Information on EC 3.5.1.93 - glutaryl-7-aminocephalosporanic-acid acylase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
3.5.1.93
-
RECOMMENDED NAME
GeneOntology No.
glutaryl-7-aminocephalosporanic-acid acylase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O = (7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis
-
-
hydrolysis
Achromobacter xylosoxidans MTCC *491, Pseudomonas sp. N176
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Penicillin and cephalosporin biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
(7R)-7-(4-carboxybutanamido)cephalosporanate amidohydrolase
Forms 7-aminocephalosporanic acid, a key intermediate in the synthesis of cephem antibiotics. It reacts only weakly with cephalosporin C.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7beta-(4-carboxybutanamido)cephalosporanic acid acylase
-
-
adipyl-cephalosporin acylase
-
-
adipyl-cephalosporin acylase
Pseudomonas sp. SY-77
-
-
-
CCA
Achromobacter xylosoxidans MTCC *491
-
-
-
cephalorin acylase
-
-
cephalosporin acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
cephalosporin acylase
-
-
cephalosporin acylase
Aeromonas sp. ACY
-
-
-
cephalosporin acylase
-
-
cephalosporin acylase
-
-
cephalosporin acylase
Brevundimonas diminuta N 176
-
-
-
cephalosporin acylase
-
-
cephalosporin acylase
Flavobacterium sp. 650
-
-
-
cephalosporin acylase
-
-
cephalosporin acylase
-
-
cephalosporin acylase
O86089
-
cephalosporin acylase
Pseudomonas sp. A14, Pseudomonas sp. BLO 72
-
-
-
cephalosporin acylase
A4ZVL3
a member of the N-terminal nucleophile hydrolase family
cephalosporin acylase
-
-
cephalosporin acylase
-
-
cephalosporin C acylase
-
-
cephalosporin C acylase
Achromobacter xylosoxidans MTCC *491
-
-
-
cephalosporin C acylase
-
-
cephalosporin C acylase
Brevibacillus laterosporus J1
-
-
-
cephalosporin C acylase
-
-
cephalosporin C acylase
Brevundimonas diminuta N176
-
-
-
cephalosporin C acylase
-
-
cephalosporin C acylase
Pseudomonas sp. N176
-
-
-
cephalosporin C acylase
-
-
cephalosporin-C acylase
-
-
CPC acylase
-
-
CPC acylase
-
-
CPC acylase
-
-
CPCAcy
-
-
GAR
-
commercial preparation
GL-7-ACA acylase
-
-
GL-7-ACA acylase
-
-
GL-7-ACA acylase
Pseudomonas nitroreducens CCRC 11041
-
-
-
GL-7-ACA acylase
O86089
-
GL-7-ACA acylase
-
-
GL-7-ACA acylase
Pseudomonas sp. KAC-1
-
;
-
GL-7ACA acylase
-
-
Gl7ACA acylase
-
-
Gl7ACA acylase
Brevundimonas diminuta VKM B-1297
-
-
-
GLA
O86089
-
glutaryl 7-amino cephalosporanic acid acylase
-
-
glutaryl 7-aminocephalosporanic acid acylase
-
-
glutaryl acylase
-
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Aeromonas sp. ACY
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Arthrobacter sp. 5-8A
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Bacillus sp. SV12
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Brevibacillus laterosporus J1
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Brevundimonas diminuta N 176
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Burkholderia cepacia BY21
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Flavobacterium sp. 650
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Pseudomonas nitroreducens CCRC 11041
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
-
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
Pseudomonas sp. A14, Pseudomonas sp. BLO 72
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
-
glutaryl-7-ACA acylase
-
glutaryl-7-ACA acylases and cephalosporin acylases have been collectively referred to as cephalosporin acylases because they all have either glutaryl-7-aminocephalosporanic acid or cephalosporin C as their substrates
glutaryl-7-ACA acylase
-
-
glutaryl-7-amino cephalosporanic acid acylase
-
-
glutaryl-7-aminocephalosporanic acid acylase
-
-
glutaryl-7-aminocephalosporanic acid acylase
Achromobacter xylosoxidans MTCC *491
-
-
-
glutaryl-7-aminocephalosporanic acid acylase
-
activity of gamma-glutamyltranspeptidase
glutaryl-7-aminocephalosporanic acid acylase
-
-
glutaryl-7-aminocephalosporanic acid acylase
-
-
glutaryl-7-aminocephalosporanic acid acylase
Pseudomonas sp. BL072, Pseudomonas sp. N176
-
-
-
glutaryl-7-aminocephalosporanic acid acylase
-
-
glutaryl-7-aminocephalosporic acid acylase
-
-
glutaryl-7-aminocephalosporic acid acylase
Brevundimonas diminuta VKM B-1297
-
-
-
J1 acylase
Brevibacillus laterosporus J1
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
56645-46-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
; strain MTCC *491
-
-
Manually annotated by BRENDA team
Achromobacter xylosoxidans MTCC *491
strain MTCC *491
-
-
Manually annotated by BRENDA team
strain ACY 95
-
-
Manually annotated by BRENDA team
Aeromonas sp. ACY
strain ACY 95
-
-
Manually annotated by BRENDA team
strain 45-8A. Activity is detected when grown at temperature below 30C, but not at higher temperatures
-
-
Manually annotated by BRENDA team
strain 5-8A
-
-
Manually annotated by BRENDA team
Arthrobacter sp. 45-8A.
strain 45-8A. Activity is detected when grown at temperature below 30C, but not at higher temperatures
-
-
Manually annotated by BRENDA team
Arthrobacter sp. 5-8A
strain 5-8A
-
-
Manually annotated by BRENDA team
strain SV12
-
-
Manually annotated by BRENDA team
Bacillus sp. SV12
strain SV12
-
-
Manually annotated by BRENDA team
Brevibacillus laterosporus J1
J1
-
-
Manually annotated by BRENDA team
Brevibacillus laterosporus J1
strain J1
-
-
Manually annotated by BRENDA team
strain N 176
-
-
Manually annotated by BRENDA team
strain VKM B-1297
-
-
Manually annotated by BRENDA team
Brevundimonas diminuta KAC-1
KAC-1
Uniprot
Manually annotated by BRENDA team
Brevundimonas diminuta N 176
strain N 176
-
-
Manually annotated by BRENDA team
Brevundimonas diminuta N176
N176
-
-
Manually annotated by BRENDA team
Brevundimonas diminuta VKM B-1297
strain VKM B-1297
-
-
Manually annotated by BRENDA team
strain BY21
-
-
Manually annotated by BRENDA team
Burkholderia cepacia BY21
strain BY21
-
-
Manually annotated by BRENDA team
strain 650
-
-
Manually annotated by BRENDA team
Flavobacterium sp. 650
strain 650
-
-
Manually annotated by BRENDA team
strain CCRC 11041
-
-
Manually annotated by BRENDA team
Pseudomonas nitroreducens CCRC 11041
CCRC 11041
-
-
Manually annotated by BRENDA team
Pseudomonas nitroreducens CCRC 11041
strain CCRC 11041
-
-
Manually annotated by BRENDA team
strain ATCC 950
-
-
Manually annotated by BRENDA team
130, recombinant enzyme
Uniprot
Manually annotated by BRENDA team
180 out of 350 clinical samples (patient samples and water, soil, and hospital environment samples) are tested for determination of cephalosporin acylase. Two strains are capable of producing cephalosporin acylase
-
-
Manually annotated by BRENDA team
A14, recombinant enzyme
-
-
Manually annotated by BRENDA team
strain 130
UniProt
Manually annotated by BRENDA team
strain 130
-
-
Manually annotated by BRENDA team
strain A14; strain BLO 72
-
-
Manually annotated by BRENDA team
strain GK16
Uniprot
Manually annotated by BRENDA team
strain KAC-1, recombinant
-
-
Manually annotated by BRENDA team
strain SY-77
-
-
Manually annotated by BRENDA team
strain V22
-
-
Manually annotated by BRENDA team
SY-77
Uniprot
Manually annotated by BRENDA team
strain A14
-
-
Manually annotated by BRENDA team
Pseudomonas sp. BL072
-
-
-
Manually annotated by BRENDA team
Pseudomonas sp. BLO 72
strain BLO 72
-
-
Manually annotated by BRENDA team
Pseudomonas sp. KAC-1
KAC-1
-
-
Manually annotated by BRENDA team
Pseudomonas sp. KAC-1
strain KAC-1, recombinant
-
-
Manually annotated by BRENDA team
Pseudomonas sp. N176
-
-
-
Manually annotated by BRENDA team
Pseudomonas sp. N176
N176
-
-
Manually annotated by BRENDA team
Pseudomonas sp. N176
strain N176
-
-
Manually annotated by BRENDA team
Pseudomonas sp. SY-77
strain SY-77
-
-
Manually annotated by BRENDA team
Pseudomonas sp. SY-77
SY-77
Uniprot
Manually annotated by BRENDA team
strain V22
-
-
Manually annotated by BRENDA team
purchased from Recordati, Milano
-
-
Manually annotated by BRENDA team
purchased from Roche
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(5R,6R)-6-[(4-carboxybutanoyl)amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid + H2O
?
show the reaction diagram
-
118% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
(6R,7R)-7-[(3-carboxypropanoyl)amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + H2O
?
show the reaction diagram
-
26.9% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
(6R,7R)-7-[(3-carboxypropanoyl)amino]-8-oxo-3-(propionyloxy)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + H2O
?
show the reaction diagram
-
2.2% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
(6R,7R)-7-[(4-carboxybutanoyl)amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + H2O
?
show the reaction diagram
-
111% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
(6R,7R)-7-[(5-carboxypentanoyl)amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + H2O
?
show the reaction diagram
-
8.3% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
(6R,7R)-7-[(5-carboxypentanoyl)amino]-8-oxo-3-(propionyloxy)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + H2O
?
show the reaction diagram
-
6.9% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas sp. N176
-
-
-
-
?
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
Achromobacter xylosoxidans MTCC *491
-
-
-
-
?
2-amino-5-[(4-nitrophenyl)amino]-5-oxopentanoic acid + H2O
?
show the reaction diagram
-
-
-
-
?
4-(glutarylamino)benzoic acid + H2O
glutarate + 4-aminobenzoate
show the reaction diagram
-
-
-
-
?
5-oxo-5-(1-phenylethoxy)pentanoic acid + H2O
glutarate + ?
show the reaction diagram
-
-
-
-
?
5-oxo-5-[(1-phenylethyl)amino]pentanoic acid + H2O
?
show the reaction diagram
-
-
-
-
?
5-[(2-methoxy-1-phenylprop-2-en-1-yl)amino]-5-oxopentanoic acid + H2O
?
show the reaction diagram
-
-
-
-
?
5-[(2-methoxy-1-phenylprop-2-en-1-yl)oxy]-5-oxopentanoic acid + H2O
?
show the reaction diagram
-
-
-
-
?
5-[(4-nitrophenyl)amino]-5-oxopentanoic acid + H2O
?
show the reaction diagram
-
-
-
-
?
5-[[(1S)-1-benzyl-2-methoxy-2-oxoethyl]amino]-5-oxopentanoic acid + H2O
?
show the reaction diagram
-
-
-
-
?
7beta-(4-carboxylbutanamino)cephalosporanic acid + H2O
7-aminocephalosporanic acid + pentanoate
show the reaction diagram
Arthrobacter sp., Arthrobacter sp. 45-8A.
-
-
-
-
?
adipyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + adipate
show the reaction diagram
Brevundimonas diminuta, Brevundimonas diminuta N176
-
activity is 19% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
adipyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + adipate
show the reaction diagram
-
-
-
-
?
adipyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + adipate
show the reaction diagram
-
activity is 0.56% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
adipyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + adipate
show the reaction diagram
Brevibacillus laterosporus, Brevibacillus laterosporus J1
-
activity is 68% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
adipyl-7-aminocephalosporanic acid + H2O
adipate + 7-aminocephalosporanate
show the reaction diagram
-
-
-
-
?
adipyl-7-aminodeacetoxycephalosporanic acid + H2O
adipate + 7-aminodeacetoxycephalosporanic acid
show the reaction diagram
-
-
-
-
?
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
deacetoxycephalosporanic acid + adipate
show the reaction diagram
P07662
-
-
-
?
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
?
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. SY-77
-
-
-
-
?
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
adipate + 7-aminodesacetoxycephalosporanate
show the reaction diagram
-
57.4% activity with 5 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
adipyl-aminopenicillanic acid + H2O
adipate + aminopenicillanic acid
show the reaction diagram
P07662
wild-type enzyme shows 70.6% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
cephalosporin C + H2O
?
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanate + 2-amino-5-hydroxypentanoate
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. N176
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
activity determined using a biological and colorimetic method. For the spectrophotometric assay p-dimethylaminobenzaldehyde is added to produce a yellow condensation product which can be measured at a wavelength of 414 nm
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Pseudomonas sp. N176
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Arthrobacter sp. 45-8A.
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
weak activity
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
low activity
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
low activity
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
O86089
low activity
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
activity is 4% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
P07662
wild-type enzyme shows 2.3% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
double mutant H296S/H309S exhibits 22fold enhanced specificity and reactivity for cephalosporin C over the natural substrate glutaryl-7-aminocephalosporanic acid
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Aeromonas sp. ACY
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Brevundimonas diminuta N176
-
activity is 4% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Pseudomonas sp. N176
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Pseudomonas sp. SY-77
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Achromobacter xylosoxidans MTCC *491
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Flavobacterium sp. 650
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Brevundimonas diminuta VKM B-1297
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Brevibacillus laterosporus J1
-
weak activity
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Brevundimonas diminuta N 176
-
-
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Pseudomonas sp. BL072
-
low activity
-
-
?
cephalosporin C + H2O
cephalosporanic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
Pseudomonas sp. BLO 72
-
-
-
-
?
cephalosporin C + H2O
7-aminocephalosporanic acid + ?
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
caphalosporamic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporamic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
-
-
-
?
cephalosporin C + H2O
cephalosporamic acid + 2-amino-5-hydroxypentanoate
show the reaction diagram
-
the recombinant enzyme exhibits 2.3times more cephalosporin C specific deacylation activity compared to glutaryl-7-aminocephalosporanic acid
-
-
?
cephalothin + H2O
?
show the reaction diagram
Brevundimonas diminuta, Brevundimonas diminuta N176
-
-
-
-
?
D-glutamyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + D-glutamate
show the reaction diagram
-
activity is 0.85% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
D-glutamyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutamate
show the reaction diagram
Brevibacillus laterosporus, Brevibacillus laterosporus J1
-
activity is 0.55% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
DGDPP + H2O
L-Asp + GDPP
show the reaction diagram
-
20% activity compared to E-GDPPDLADQG
-
-
?
dimethyl glutarate + (2S)-2-amino-3-(4-hydroxyphenyl)propanamide
5-[[(1S)-2-amino-1-(4-hydroxybenzyl)-2-oxoethyl]amino]-5-oxopentanoic acid + ?
show the reaction diagram
-
the enzyme catalyzes the synthesis of the amide bond when working at very high substrate concentrations, namely in heterogeneous substrate mixtures, whereas no enzymatic activity is dtected using the enzyme in diluted organic solvent solutions. A negative charge on the acyl moiety is crucial for substrate recognition
-
-
?
dimethyl glutarate + ethyl (2S)-2-amino-3-(4-hydroxyphenyl)propanoate
5-[[(1S)-2-ethoxy-1-(4-hydroxybenzyl)-2-oxoethyl]amino]-5-oxopentanoic acid + ?
show the reaction diagram
-
the enzyme catalyzes the synthesis of the amide bond when working at very high substrate concentrations, namely in heterogeneous substrate mixtures, whereas no enzymatic activity is detected using the enzyme in diluted organic solvent solutions. A negative charge on the acyl moiety is crucial for substrate recognition
-
-
?
EGDPP + H2O
L-Glu + GDPP
show the reaction diagram
-
67% activity compared to E-GDPPDLADQG
-
-
?
EGDPPDLADQG + H2O
L-Glu + GDPPDLADQG
show the reaction diagram
-
100% activity
-
-
?
gamma-Glu-Cys-Gly
gamma-Glu + Cys-Gly
show the reaction diagram
-
-
-
-
-
GEGDPP + H2O
Gly-L-Glu + GDPP
show the reaction diagram
-
5% activity compared to E-GDPPDLADQG
-
-
?
glutaryl 7-aminodeacetoxycephalosporanic acid + H2O
7-aminodeacetoxycephalosporanic acid + glutarate
show the reaction diagram
O86089
-
-
-
?
glutaryl deacetoxy 7-aminocephalosporanic acid + H2O
deacetoxycephalosporanic acid + glutarate
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. BL072
-
-
-
-
?
glutaryl deacetyl-7-aminocephalosporanic acid + H2O
deacetyl-7-aminocephalosporanic acid + glutarate
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. BL072
-
-
-
-
?
glutaryl-3-chloro-7-aminodesacetoxycephalosporanic acid + H2O
glutarate + 3-chloro-7-aminodesacetoxycephalosporanate
show the reaction diagram
-
81.9% activity with 5 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-7-amino cephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. N176
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
P07662
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
P07662
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Q9L5D6
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
O86089
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
O86089
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
100% activity
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
rate of synthesis is fold slower than rate of synthesis
-
-
r
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
rate of synthesis is slower than rate of hydrolysis
-
-
r
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Aeromonas sp. ACY
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Brevundimonas diminuta N176
-
rate of synthesis is fold slower than rate of synthesis
-
-
r
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Burkholderia cepacia BY21, Arthrobacter sp. 5-8A
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas sp. N176
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas sp. KAC-1
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas sp. SY-77
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Flavobacterium sp. 650
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Brevundimonas diminuta VKM B-1297
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Brevibacillus laterosporus J1
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Brevibacillus laterosporus J1
-
rate of synthesis is slower than rate of hydrolysis
-
-
r
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Brevundimonas diminuta N 176
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas nitroreducens CCRC 11041
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas sp. BL072
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + glutarate
show the reaction diagram
Bacillus sp. SV12, Pseudomonas sp. BLO 72
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutarate
show the reaction diagram
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutarate
show the reaction diagram
-
activity assay
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutarate
show the reaction diagram
-
double mutant H296S/H309S exhibits 22fold enhanced specificity and reactivity for cephalosporin C over the natural substrate glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutarate
show the reaction diagram
Pseudomonas sp. N176
-
activity assay
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutarate
show the reaction diagram
Pseudomonas sp. N176
-
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
glutarate + 7-aminocephalosporanate
show the reaction diagram
-
9.9% activity with 5 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-7-aminodesacetoxycephalosporanic acid + H2O
glutarate + 7-aminodesacetoxycephalosporanate
show the reaction diagram
-
71.5% activity with 5 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-alanyl-proline + H2O
glutarate + alanyl-proline
show the reaction diagram
-
-
-
-
?
glutaryl-aminopenicillanic acid + H2O
glutarate + aminopenicillanic acid
show the reaction diagram
P07662
wild-type enzyme shows 90.6% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-D-Phe-Gly + H2O
glutarate + D-Phe-Gly
show the reaction diagram
-
about 25% activity with 50 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-deacetoxy-7-aminocephalosporanic acid + H2O
deacetoxycephalosporanic acid + glutarate
show the reaction diagram
Pseudomonas nitroreducens, Pseudomonas nitroreducens CCRC 11041
-
activity is 2.94fold higher than with glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-EGDPPDLADQG + H2O
glutarate + EGDPPDLADQG
show the reaction diagram
-
-
-
-
?
glutaryl-gamma-Glu-Cys-Gly + H2O
glutarate + gamma-Glu-Cys-Gly
show the reaction diagram
-
-
-
-
?
glutaryl-GDAADAADKG + H2O
glutarate + GDAADAADKG
show the reaction diagram
-
-
-
-
?
glutaryl-GDPP + H2O
glutarate + GDPP
show the reaction diagram
-
substrate with highest catalytic efficiency
-
-
?
glutaryl-GDPPDLADQG + H2O
glutarate + GDPPDLADQG
show the reaction diagram
-
-
-
-
?
glutaryl-GGGGAA + H2O
glutarate + GGGGAA
show the reaction diagram
-
-
-
-
?
glutaryl-GGGGGK + H2O
glutarate + GGGGGK
show the reaction diagram
-
-
-
-
?
glutaryl-glycine + H2O
glutarate + glycine
show the reaction diagram
-
-
-
-
?
glutaryl-glycine + H2O
glutaryl + glycine
show the reaction diagram
-
less than 5% activity with 50 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-glycyl-proline + H2O
glutarate + glycyline-proline
show the reaction diagram
-
-
-
-
?
glutaryl-L-Ala + H2O
glutarate + L-Ala
show the reaction diagram
-
about 25% activity with 50 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-L-Phe + H2O
glutarate + L-Phe
show the reaction diagram
-
about 10% activity with 50 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-L-Phe-Gly + H2O
glutarate + L-Phe-Gly
show the reaction diagram
-
about 50% activity with 50 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
glutaryl-tryptophan + H2O
glutarate + tryptophan
show the reaction diagram
-
-
-
-
?
glutarylaniline + H2O
glutarate + aniline
show the reaction diagram
-
-
-
-
?
L-gamma-glutamyl p-nitroanilide + H2O
L-glutamate + p-nitroaniline
show the reaction diagram
-
-
-
-
-
L-gamma-glutamyl p-nitroanilide + H2O
L-glutamate + p-nitroaniline
show the reaction diagram
-
-
-
-
?
L-gamma-glutamyl p-nitroanilide + H2O
L-glutamate + p-nitroaniline
show the reaction diagram
-
-
-
-
-
N-acetyl cephalosporin C + H2O
acetate + cephalosporin C
show the reaction diagram
Brevundimonas diminuta, Brevundimonas diminuta N176
-
activity is 1% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-D-alanine methyl ester + H2O
glutarate + D-alanine methyl ester
show the reaction diagram
-
2.2% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-D-phenylalanine + H2O
glutarate + D-Phe
show the reaction diagram
-
2% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-D-phenylalanine methyl ester + H2O
glutarate + D-phenylalanine methyl ester
show the reaction diagram
-
4.5% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-D-phenylglycine + H2O
glutarate + D-phenylglycine
show the reaction diagram
-
1.1% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-D-phenylglycine methyl ester + H2O
glutarate + D-phenylglycine methyl ester
show the reaction diagram
-
0.23% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-L-Ala + H2O
glutarate + L-Ala
show the reaction diagram
-
2.7% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-L-alanine methyl ester + H2O
glutarate + L-alanine methyl ester
show the reaction diagram
-
25% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-L-phenylalanine + H2O
glutarate + L-Phe
show the reaction diagram
-
2.5% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-L-phenylalanine methyl ester + H2O
glutarate + L-phenylalanine methyl ester
show the reaction diagram
-
15.7% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-L-phenylglycine + H2O
glutarate + L-phenylglycine
show the reaction diagram
-
7.5% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutaryl-L-phenylglycine methyl ester + H2O
glutarate + L-phenylglycine methyl ester
show the reaction diagram
-
59.3% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
N-glutarylglycine methyl ester + H2O
glutarate + glycine methyl ester
show the reaction diagram
-
9.7% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
penicillin G + H2O
?
show the reaction diagram
O86089
low activity
-
-
?
pentanoyl-7-aminocephalosporanic acid + H2O
pentanoate + 7-aminocephalosporanate
show the reaction diagram
-
about 2.4% activity with 5 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
succinyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + succinate
show the reaction diagram
-
-
-
-
?
succinyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanate + succinate
show the reaction diagram
-
activity is 32% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
succinyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + succinate
show the reaction diagram
-
activity is 6.1% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
succinyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + succinate
show the reaction diagram
-
activity is 9.9fold higher than with glutaryl-7-aminocephalosporanic acid
-
-
?
succinyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + succinate
show the reaction diagram
Brevibacillus laterosporus J1
-
activity is 6.1% of the activity with glutaryl-7-aminocephalosporanic acid
-
-
?
succinyl-7-aminocephalosporanic acid + H2O
succinate + 7-aminocephalosporanate
show the reaction diagram
-
25.8% activity with 5 mM substrate compared to glutaryl-7-aminocephalosporanic acid
-
-
?
L-gamma-glutamyl p-nitroanilide + H2O
L-glutamate + p-nitroaniline
show the reaction diagram
Pseudomonas nitroreducens CCRC 11041
-
-
-
-
?
additional information
?
-
-
enzyme also exhibits L-gamma-glutamyltranspeptidase activity
-
-
-
additional information
?
-
-
no activity with cephalosporin C and succinyl-4-aminocephalosporanic acid
-
-
-
additional information
?
-
-
purified enzyme exhibits also gamma-glutamyltranspeptidase activity with L-gamma-glutamyl-p-nitroanilide
-
-
-
additional information
?
-
-
the enzyme has transacylase activity 10times that of its hydrolytic activity
-
-
-
additional information
?
-
-
less than 1% activity with TLGEGDPP compared to EGDPPDLADQG, no activity with adipyl-7-aminocephalosporanic acid, the enzyme also removes N-terminal Glu or Asp from peptides. Cephalosporin acylase is an acylpeptide hydrolase rather than a previously expected acylamino acid acylase. It only exhibits exopeptidase activity for the hydrolysis of an externally added peptide, supporting the intra-molecular interaction, the precursor cephalosporin acylase (without signal peptide) is activated by two peptide bond cleavages, which excise an internal spacer. The first cleavage occurs between Gly169 and Ser170 and the second cleavage occurs between Glu159 and Gly160
-
-
-
additional information
?
-
-
the recombinant enzyme shows no activity towards cephalosporin C, butanoyl-7-aminodesacetoxycephalosporanic acid and pentanoyl-7-aminodesacetoxycephalosporanic acid
-
-
-
additional information
?
-
-
purified enzyme exhibits also gamma-glutamyltranspeptidase activity with L-gamma-glutamyl-p-nitroanilide
-
-
-
additional information
?
-
Pseudomonas nitroreducens CCRC 11041
-
enzyme also exhibits L-gamma-glutamyltranspeptidase activity
-
-
-
additional information
?
-
Pseudomonas nitroreducens CCRC 11041
-
no activity with cephalosporin C and succinyl-4-aminocephalosporanic acid
-
-
-
additional information
?
-
Pseudomonas sp. BL072
-
the enzyme has transacylase activity 10times that of its hydrolytic activity
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
-
-
-
-
?
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
Pseudomonas sp. N176
-
-
-
-
?
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
(7R)-7-aminocephalosporanate + glutarate
show the reaction diagram
Achromobacter xylosoxidans MTCC *491
-
-
-
-
?
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
7-aminocephalosporanate
-
20 mM, 81% inhibition
7-aminocephalosporanic acid
-
product inhibition, competitive inhibitor
7beta-3-bromopropionyl aminocephalosporanic acid
-
Trp4 of the beta-subunit is alkylated
7beta-bromoacetyl amino cephalosporanic acid
-
inhibits and alkylates the enzyme. The enzyme labeled with 7beta-bromoacetyl amino cephalosporanic acid is inactive at room temperature, but in the process of crystallization at 18C it catalyzes the hydrolysis of 7beta-bromoacetyl amino cephalosporanic acid.. In crystals, 7-aminocephalosporanic acid is released but the acetic acid still binds with Trp-beta4, and as a result, the enzyme remains inactive
7beta-bromoacetyl aminocephalosporanic acid
-
Trp4 of the beta-subunit is alkylated
cephalosporin C
-
20 mM, 28% inhibition
EDTA
-
5 mM, 27% inhibition
Glutaric acid
-
20 mM, 28% loss of activity
PCMB
-
92% inhibition at 1 mM, 96% inhibition at 1 mM
PCMB
-
5 mM, 10% inhibition
PMSF
-
10% inhibition at 1 mM, 22% inhibition at 5 mM
PMSF
-
5 mM, 38% inhibition
succinyl-7-aminocephalosporanic acid
-
20 mM, 23% loss of activity
glutaryl-7-aminocephalosporanic acid
-
substrate inhibition
additional information
-
nitrogen sources have a repressive effect on enzyme activity
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
D-glucose
-
9% increase of activity in the presence of glucose
galactose
-
22% increase of activity in the presence of galactose
N-isobutoxycarbomyl cephalosporin C
-
-
monopotassium anthroquinone 1,8-di-sulfonate
-
-
additional information
A4ZVL3
is activated through two steps of intramolecular autoproteolysis, the first mediated by a serine residue, and the second by a glutamate, which releases the pro-segment and produces the active enzyme
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.16
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375G
0.17
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375C
0.51
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375A
0.65
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375M
0.7
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375L; pH 7.5, 37C, mutant enzyme F375Y
0.83
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375N
0.9
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375H; pH 7.5, 37C, mutant enzyme F375T
0.99
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375S
1
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, wild-type enzyme
1.37
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375V
1.5
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375W
2.07
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375I
2.1
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375Q
2.16
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375K
4.3
-
adipyl-7-aminocephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
4.4
-
adipyl-7-aminocephalosporanic acid
-
recombinant enzyme, at pH 8.0 and 25C
7.5
-
adipyl-7-aminocephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
10
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375R
11
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375D
12
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375E
0.14
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266H
0.33
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266M
0.59
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266W
0.6
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266F
0.62
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266Q
0.65
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266Y
0.8
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266C
0.99
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266S
1
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266L
1.2
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
wild-type enzyme
1.4
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266G
2.2
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266A; mutant enzymeN266D
4.4
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266P
5.7
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266T
13.8
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzymeN266E
0.14
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266H
0.14
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266H
0.17
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266Q/F375L; mutant enzyme Y178H/N266M
0.19
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme Y178H/N266H
0.28
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme Y178H/N266Q/F375L
0.33
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266M
0.42
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266S
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme Y178H
0.52
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F229L
0.62
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266Q
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme F375M
0.7
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme M271V/Q291K/T374S
0.7
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme F375L
0.7
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
0.8
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
wild type enzyme
0.82
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F375L
0.96
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, wild-type enzyme
3.8
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
recombinant enzyme, at pH 8.0 and 25C
7.9
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
1.7
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H296S/H417M
2.7
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme Y271V
3.2
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215E
4.1
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H296F
4.8
-
cephalosporin C
-
-
4.8
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H296T
4.8
-
cephalosporin C
-
-
5.2
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H296N
6.3
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H309Y
6.5
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme F270M
6.7
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H296S
6.9
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215Y
7.2
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme D416Y
7.7
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215F
8
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme Y271F
8.3
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215V
9.5
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215L
9.5
-
cephalosporin C
-
wild type enzyme, at pH 8.0 and 25C
10
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215Y/H296S
10.1
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme Y215Y/F270S
11
-
cephalosporin C
-
pH 8.0, 37C, wild-type enzyme
12.2
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H296S/H309S
12.2
-
cephalosporin C
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
12.3
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H417Y
13
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme D416Y/H417Y
17.2
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215Y/H309S
17.8
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme A215Y/H296S/H309S
18.9
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme E89A/A215Y
21.3
-
cephalosporin C
-
37C, mutant enzyme Y269F
21.6
-
cephalosporin C
-
37C, wild-type enzyme
22
-
cephalosporin C
-
at 37C, pH not specified in the publication
22.1
-
cephalosporin C
-
37C, mutant enzyme Y269Y
24.4
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme H309S
24.5
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme S22P/T394P/D416Y/H417Y
27.8
-
cephalosporin C
-
37C, mutant enzyme Y270F
32
-
cephalosporin C
-
pH 8.0, 37C, mutant enzyme I44V/E49stop/D416Y/H417Y
37
-
cephalosporin C
-
mutant enzyme L677F, in 100 mM Tris-HCl buffer, pH 8.0, temperature not specified in the publication
43
-
cephalosporin C
-
wild type enzyme, in 100 mM Tris-HCl buffer, pH 8.0, temperature not specified in the publication
86
-
cephalosporin C
-
mutant enzyme P295A, in 100 mM Tris-HCl buffer, pH 8.0, temperature not specified in the publication
105
-
cephalosporin C
-
mutant enzyme A675G, in 100 mM Tris-HCl buffer, pH 8.0, temperature not specified in the publication
121
-
cephalosporin C
-
mutant enzyme H309V, in 100 mM Tris-HCl buffer, pH 8.0, temperature not specified in the publication
3.3
-
gamma-Glu-Cys-Gly
-
at 37C, pH not specified in the publication
-
1.6
-
glutaryl deacetoxy 7-aminocephalosporanic acid
-
-
1
-
glutaryl-3-chloro-7-aminodesacetoxycephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
-
2.2
-
glutaryl-3-chloro-7-aminodesacetoxycephalosporanic acid
-
recombinant enzyme, at pH 8.0 and 25C
-
8.7
-
glutaryl-3-chloro-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
-
0.7
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H296S
0.9
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme A215F; pH 8.0, 37C, mutant enzyme A215Y/H309S
1
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme A215E; pH 8.0, 37C, mutant enzyme H309Y
1.1
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme E89A/A215Y
1.2
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme A215V; pH 8.0, 37C, mutant enzyme Y271F
1.6
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
1.7
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme A215L; pH 8.0, 37C, mutant enzyme A215Y; pH 8.0, 37C, mutant enzyme D416Y
1.9
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H296F; pH 8.0, 37C, mutant enzyme Y27V
2
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H296T; pH 8.0, 37C, mutant enzyme Y215Y/F270S
2.5
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme A215Y/H296S/H309S
2.7
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme F270M
2.8
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H296N
4
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme A215Y/H296S
4.3
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H417Y
4.6
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H309S
5.5
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H296S/H417M
6.9
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme H296S/H309S
9
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme D416Y/H417Y
25
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme I44V/E49stop/D416Y/H417Y
37.2
-
glutaryl-7-amino cephalosporanic acid
-
pH 8.0, 37C, mutant enzyme S22P/T394P/D416Y/H417Y
0.016
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266H
0.031
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, wild-type enzyme
0.031
-
glutaryl-7-aminocephalosporanic acid
-
wild-type enzyme
0.033
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, wild-type enzyme
0.038
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375C
0.04
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266H
0.042
-
glutaryl-7-aminocephalosporanic acid
-
wild type enzyme
0.043
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme M271V/Q291K/T374S
0.044
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266H
0.048
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F229L
0.066
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375G
0.07
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375A; pH 7.5, 37C, mutant enzyme F375S
0.075
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266S
0.08
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375Y
0.08
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme Y178H; mutant enzyme Y178H/N266H; mutant enzyme Y178H/N266Q/F375L
0.09
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375T
0.094
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266S
0.1
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375I; pH 7.5, 37C, mutant enzyme F375V
0.1
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266F
0.11
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375H
0.11
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266Y
0.12
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266Q
0.12
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266Q
0.15
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, mutant enzyme Y126F
0.16
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266C
0.17
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375M
0.17
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266M
0.17
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme F375M; mutant enzyme N266M; mutant enzyme N266Q/F375L
0.18
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375N
0.18
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266W
0.19
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
0.2
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375E
0.21
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
0.22
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375D
0.26
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375L
0.26
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266A
0.26
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme F375L
0.27
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F375L
0.29
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266E
0.3
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme Y178H/N266M
0.33
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4A; pH 8.0, 37C, beta-subunit mutant W4Y
0.36
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4T
0.37
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4F; pH 8.0, 37C, beta-subunit mutant W4L
0.4
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4H
0.43
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266L
0.46
-
glutaryl-7-aminocephalosporanic acid
-
-
0.5
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, wild-type enzyme
0.5
-
glutaryl-7-aminocephalosporanic acid
-
at 37C, pH not specified in the publication
0.54
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266G
0.6
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375R
0.7
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375K; pH 7.5, 37C, mutant enzyme F375Q
0.8
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375W
0.82
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme H23Q
1.18
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme E455Q
1.2
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, mutant enzyme W173A
1.24
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme E455D
1.3
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266P
1.5
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266T
1.5
-
glutaryl-7-aminocephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
1.69
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, mutant enzyme Y126A
1.76
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme H23D
2.1
-
glutaryl-7-aminocephalosporanic acid
-
-
2.5
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164A
2.5
-
glutaryl-7-aminocephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
2.57
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164G
2.6
-
glutaryl-7-aminocephalosporanic acid
-
-
3
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
3.1
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunit mutant enzyme E455L
3.2
-
glutaryl-7-aminocephalosporanic acid
-
-
3.3
-
glutaryl-7-aminocephalosporanic acid
-
recombinant enzyme, at pH 8.0 and 25C
3.68
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164N
5.4
-
glutaryl-7-aminocephalosporanic acid
-
recombinant enzyme, at pH 8.0 and 25C
5.85
-
glutaryl-7-aminocephalosporanic acid
-
37C, wild-type enzyme
6.1
-
glutaryl-7-aminocephalosporanic acid
-
-
6.9
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
9.32
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164Q
9.9
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, soluble enzyme
10.41
-
glutaryl-7-aminocephalosporanic acid
-
-
11.7
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme K198A
12.2
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme R121A
12.3
-
glutaryl-7-aminocephalosporanic acid
P07662
wild-type enzyme
12.7
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme D286A
13.4
-
glutaryl-7-aminocephalosporanic acid
P07662
mutant enzyme with mutation Y151F in alpha-subunit and mutation Q50N in beta-subunit
14.4
-
glutaryl-7-aminocephalosporanic acid
P07662
alpha-subunit mutant enzyme Y151F
15.4
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane
17.8
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme Q50N/K198A
17.9
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme Q50N
1.3
-
glutaryl-7-aminodesacetoxycephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
-
1.5
-
glutaryl-7-aminodesacetoxycephalosporanic acid
-
recombinant enzyme, at pH 8.0 and 25C
-
14.1
-
glutaryl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
-
0.73
-
glutaryl-alanyl-proline
-
at 37C, pH not specified in the publication
-
2.7
-
glutaryl-EGDPPDLADQG
-
at 37C, pH not specified in the publication
-
6.4
-
glutaryl-gamma-Glu-Cys-Gly
-
at 37C, pH not specified in the publication
-
0.4
-
glutaryl-GDAADAADKG
-
at 37C, pH not specified in the publication
-
0.35
-
glutaryl-GDPP
-
at 37C, pH not specified in the publication
-
0.67
-
glutaryl-GDPPDLADQG
-
at 37C, pH not specified in the publication
-
1.68
-
glutaryl-GGGGAA
-
at 37C, pH not specified in the publication
-
0.43
-
glutaryl-GGGGGK
-
at 37C, pH not specified in the publication
-
3.1
-
glutaryl-glycine
-
at 37C, pH not specified in the publication
-
0.85
-
glutaryl-glycyl-proline
-
at 37C, pH not specified in the publication
-
6.1
-
glutaryl-tryptophan
-
at 37C, pH not specified in the publication
-
40
-
glutarylaniline
-
at 37C, pH not specified in the publication
5.92
-
L-gamma-glutamyl p-nitroanilide
-
-
16.1
-
pentanoyl-7-aminocephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
-
17.4
-
pentanoyl-7-aminocephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
-
2.8
-
succinyl-7-aminocephalosporanic acid
-
recombinant enzyme, at pH 8.0 and 25C; wild type enzyme, at pH 8.0 and 25C
3.2
-
succinyl-7-aminocephalosporanic acid
-
at 37C, pH not specified in the publication
5.4
-
succinyl-7-aminocephalosporanic acid
-
mutant enzyme H296S/H309S, at pH 8.0 and 25C
4.5
-
4-(glutarylamino)benzoic acid
-
at 37C, pH not specified in the publication
-
additional information
-
7-aminocephalosporanic acid
-
estimated value: 9.43 mg/ml, pH 7.6, 34C, Vmax: 7.65 U/ml
3.8
-
L-gamma-glutamyl-p-nitroanilide
-
-
additional information
-
additional information
-
the apparent Km value of CPC acylase increases and Vmax is almost unaffected in the presence of 7-aminocephalosporanic acid
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.16
-
4-(glutarylamino)benzoic acid
-
at 37C, pH not specified in the publication
-
0.0485
-
7-aminocephalosporanic acid
-
pH and temperature not specified in the publication
0.508
-
7-aminocephalosporanic acid
-
D445G mutant, pH and temperature not specified in the publication
0.015
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375D
0.018
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375R
0.026
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375W
0.036
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375K
0.073
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375G
0.08
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375E
0.09
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375V
0.097
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375I
0.11
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375S
0.2
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375A
0.24
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375T
0.43
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375Y
0.49
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, wild-type enzyme
0.55
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375C
0.65
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375M
0.67
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375L
0.68
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375Q
0.9
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375N
1.16
-
adipyl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375H
0.002
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266D
0.023
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266E
0.061
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266P
0.15
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266T
0.18
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266A
0.23
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266G
0.24
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266S
0.25
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266L
0.31
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266C
0.33
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266W; mutant enzyme N266Y
0.37
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266F
0.41
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
wild-type enzyme
0.46
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266Q
0.47
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266H
1.9
-
adipyl-7-aminodeacetoxycephalosporanic acid
-
mutant enzyme N266M
0.34
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266S
0.38
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
wild type enzyme
0.46
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, wild-type enzyme
0.46
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266Q
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F229L
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266H
0.48
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266Q/F375L
0.55
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266H
0.61
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme M271V/Q291K/T374S
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme F375M
0.66
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme Y178H; mutant enzyme Y178H/N266Q/F375L
0.67
-
adipyl-7-aminodesacetoxycephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F375L
0.67
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme F375L
0.95
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme Y178H/N266H
1.2
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme Y178H/N266M
1.9
-
adipyl-7-aminodesacetoxycephalosporanic acid
-
mutant enzyme N266M
0.015
-
cephalosporin C
-
at 37C, pH not specified in the publication
2.91
-
cephalosporin C
-
37C, mutant enzyme Y270F
5.93
-
cephalosporin C
-
37C, wild-type enzyme
9.4
-
cephalosporin C
-
37C, mutant enzyme Y269Y
10.4
-
cephalosporin C
-
37C, mutant enzyme Y269F
0.008
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375E
0.01
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375D
0.025
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266E
0.031
0.51
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
0.082
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266L
0.13
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266P
0.21
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme Q50N/K198A
0.21
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375R
0.22
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme Q50N
0.23
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, mutant enzyme W173A
0.24
-
glutaryl-7-aminocephalosporanic acid
P07662
alpha-subunit mutant enzyme Y151F
0.38
-
glutaryl-7-aminocephalosporanic acid
P07662
mutant enzyme with mutation Y151F in the alpha-subunit and Q50N in the beta-subunit
0.41
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme D286A
0.45
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme R121A; wild-type enzyme
0.47
-
glutaryl-7-aminocephalosporanic acid
P07662
beta-subunit mutant enzyme K198betaA
0.47
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375W
0.49
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266T
0.61
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266Y
0.67
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375I
0.75
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266Q/F375L
0.8
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375Q
0.89
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme Y178H/N266M
0.9
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266F
1.02
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375V
1.1
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375L
1.1
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F375L
1.1
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme F375L
1.3
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266W
1.334
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375H
1.4
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266A
1.6
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266S
1.8
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375T
1.8
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266M
1.8
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266M
1.96
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375N
2
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266C; mutant enzyme N266G
2.02
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375M
2.02
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme F375M
2.2
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375K
2.2
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266S
2.2
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme Y178H/N266H
2.308
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375C
2.35
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375A
2.4
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266Q
2.4
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266Q
2.46
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375G
2.5
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme Y178H; mutant enzyme Y178H/N266Q/F375L
3.1
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375S
3.1
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266H
3.1
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme F229L
3.1
-
glutaryl-7-aminocephalosporanic acid
-
mutant enzyme N266H
3.3
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4T
3.3
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme N266H
3.4
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme E455L
3.5
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, mutant enzyme M271V/Q291K/T374S
3.6
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4A
3.64
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, mutant enzyme F375Y
3.8
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4L
4
-
glutaryl-7-aminocephalosporanic acid
-
pH 7.5, 37C, wild-type enzyme
4
-
glutaryl-7-aminocephalosporanic acid
-
wild-type enzyme
4.1
-
glutaryl-7-aminocephalosporanic acid
-
wild type enzyme
4.3
-
glutaryl-7-aminocephalosporanic acid
P07662
pH 7.5, 37C, wild-type enzyme
5
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4F
5.5
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4H
5.9
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4Y
6.05
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, mutant enzyme Y126F
6.1
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme H23D
6.5
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunit mutant enzyme H23Q
6.6
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme E455Q
7.1
-
glutaryl-7-aminocephalosporanic acid
-
at 37C, pH not specified in the publication
7.16
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, mutant enzyme Y126A
7.54
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
7.8
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
8.29
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164Q
9.4
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, beta-subunitmutant enzyme E455D
15.1
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164N
16.9
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 30C, wild-type enzyme
18.1
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164A
18.6
-
glutaryl-7-aminocephalosporanic acid
-
37C, mutant enzyme M164G
49.8
-
glutaryl-7-aminocephalosporanic acid
-
37C, wild-type enzyme
6.2
-
glutaryl-alanyl-proline
-
at 37C, pH not specified in the publication
-
12.8
-
glutaryl-EGDPPDLADQG
-
at 37C, pH not specified in the publication
-
3.2
-
glutaryl-gamma-Glu-Cys-Gly
-
at 37C, pH not specified in the publication
-
4.8
-
glutaryl-GDAADAADKG
-
at 37C, pH not specified in the publication
-
13.8
-
glutaryl-GDPP
-
at 37C, pH not specified in the publication
-
13.2
-
glutaryl-GDPPDLADQG
-
at 37C, pH not specified in the publication
-
1.36
-
glutaryl-GGGGAA
-
at 37C, pH not specified in the publication
-
1.73
-
glutaryl-GGGGGK
-
at 37C, pH not specified in the publication
-
1.65
-
glutaryl-glycine
-
at 37C, pH not specified in the publication
-
5.4
-
glutaryl-glycyl-proline
-
at 37C, pH not specified in the publication
-
3.1
-
glutaryl-tryptophan
-
at 37C, pH not specified in the publication
-
0.35
-
glutarylaniline
-
at 37C, pH not specified in the publication
1.28
-
succinyl-7-aminocephalosporanic acid
-
at 37C, pH not specified in the publication
0.33
-
gamma-Glu-Cys-Gly
-
at 37C, pH not specified in the publication
-
additional information
-
glutaryl-7-aminocephalosporanic acid
-
pH 8.0, 37C, mutant enzyme Y126F
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.04
-
4-(glutarylamino)benzoic acid
-
at 37C, pH not specified in the publication
0
3.41
-
7-aminocephalosporanic acid
-
E423/E442Q/D445N mutant, pH and temperature not specified in the publication
5842
0.00068
-
cephalosporin C
-
at 37C, pH not specified in the publication
8411
0.1
-
gamma-Glu-Cys-Gly
-
at 37C, pH not specified in the publication
0
14.2
-
glutaryl-7-aminocephalosporanic acid
-
at 37C, pH not specified in the publication
213015
8.5
-
glutaryl-alanyl-proline
-
at 37C, pH not specified in the publication
0
4.7
-
glutaryl-EGDPPDLADQG
-
at 37C, pH not specified in the publication
0
0.5
-
glutaryl-gamma-Glu-Cys-Gly
-
at 37C, pH not specified in the publication
0
12
-
glutaryl-GDAADAADKG
-
at 37C, pH not specified in the publication
0
40
-
glutaryl-GDPP
-
at 37C, pH not specified in the publication
0
20
-
glutaryl-GDPPDLADQG
-
at 37C, pH not specified in the publication
0
0.81
-
glutaryl-GGGGAA
-
at 37C, pH not specified in the publication
0
4
-
glutaryl-GGGGGK
-
at 37C, pH not specified in the publication
0
0.53
-
glutaryl-glycine
-
at 37C, pH not specified in the publication
0
6.4
-
glutaryl-glycyl-proline
-
at 37C, pH not specified in the publication
0
0.56
-
glutaryl-tryptophan
-
at 37C, pH not specified in the publication
0
0.01
-
glutarylaniline
-
at 37C, pH not specified in the publication
324836
0.4
-
succinyl-7-aminocephalosporanic acid
-
at 37C, pH not specified in the publication
222400
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.4
-
7-aminocephalosporanate
-
-
2
-
7-aminocephalosporanate
-
-
2.3
-
7-aminocephalosporanate
-
-
14
-
7beta-3-bromopropionyl aminocephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
20
-
7beta-3-bromopropionyl aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4Y
0.87
-
7beta-bromoacetyl aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4Y
5.4
-
7beta-bromoacetyl aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4H; pH 8.0, 37C, beta-subunit mutant W4T
5.5
-
7beta-bromoacetyl aminocephalosporanic acid
-
pH 8.0, 37C, wild-type enzyme
6.3
-
7beta-bromoacetyl aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4F
6.4
-
7beta-bromoacetyl aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4A
6.8
-
7beta-bromoacetyl aminocephalosporanic acid
-
pH 8.0, 37C, beta-subunit mutant W4L
2.5
-
Glutarate
-
-
10
-
Glutarate
-
-
21
-
glutaryl-7-aminocephalosporanic acid
-
wild type enzyme, at pH 8.0 and 25C
0.102
-
PCMB
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.28
-
-
LB Miller broth medium
1.6
-
-
crude extract, at 37C
3.7
-
-
purification step crude extract
10
-
-
after 6fold purification, at 37C
11.3
-
P07662
beta-subunit mutant enzyme K198A
11.4
-
-
optimized conditions, SB3 medium, 12.5 g NaCl/l, 0.6 mM IPTG
11.6
-
P07662
beta-subunit mutant enzyme R121A
11.8
-
P07662
wild-type enzyme
12
-
P07662
beta-subunit mutant enzyme D286A
12.3
-
P07662
mutant enzyme with mutation Y151F in alpha-subunit and mutation Q50N in beta-subunit
14.4
-
P07662
alpha-subunit mutant enzyme Y151F
16.6
-
-
-
17.4
-
P07662
beta-subunit mutant enzyme Q50N/K198A
17.5
-
P07662
beta-subunit mutant enzyme Q50N
125
-
-
purification step HiTrap chelating chromatography
133
-
-
purification step PD10 desalting
additional information
-
-
it is possible to immobilize up to 85 IU/g at 25C that corresponds to 200 IU/g at 37C
additional information
-
-
6810 U/l in a 5l fermentor
additional information
-
-
double mutant H296S/H309S exhibits 22fold enhanced specificity and reactivity for cephalosporin C over the natural substrate glutaryl-7-aminocephalosporanic acid
additional information
-
-
immobilized enzyme, on LX-1000EP support, 81 U/g wet resin; purified enzyme 10 U/mg wet resin
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
-
P07662
beta-subunit mutant enzyme R121A
7
-
P07662
wild-type enzyme
7.5
-
-
enzyme immobilized on cyanogen bromide agarose, enzyme immobilized on amino-epoxy Sepabead derivatives and enzyme and soluble enzyme
7.6
-
-
for the 7-aminocephalosporanic acid production
8
8.5
-
soluble enzyme and enzyme immobilized on silica gel modified by epoxide silanization
8
-
P07662
beta-subunit mutant enzyme K198A
8
-
-
soluble enzyme
8
-
-
reaction with glutaryl-7-aminocephalosporanic acid
8
-
-
assay at
8.3
-
-
enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane
8.5
9.5
-
LX-1000EP-immobilized enzyme
8.5
9.5
-
broad pH optimum, immobilized enzyme
8.5
-
-
free enzyme
9
10
-
reaction with glutaryl-7-aminocephalosporanic acid
9
-
P07662
beta-subunit mutant enzyme D286A
9
-
-
reaction with glutaryl-7-aminocephalosporanic acid
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
6
-
pH 5.0: about 70% of maximal activity, pH 6.0: about 75% of maximal activity, pH 4.5: about 20% of maximal activity, pH 6.5: about 15% of maximal activity
5
6
-
pH 4.0: no activity, pH 5.0: maximal activity, pH 6.0: about 60% of maximal activity, pH 7: no activity
6
10
-
pH 6.0: soluble enzyme shows 35% of maximal activity, enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane shows 60% of maximal activity, pH 10.0: soluble enzyme shows 67% of maximal activity, enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane shows 75% of maximal activity
6
10
-
pH 6.0: soluble enzyme shows about 35% of maximal activity, enzyme immobilized on silica gel modified by epoxide silanization shows about 65% of maximal activity, pH 10.0: soluble enzyme shows about 70% of maximal activity, enzyme immobilized on silica gel modified by epoxide silanization shows about 85% of maximal activity
6
9
-
pH 6.0: about 40% of maximal activity of enzyme immobilized on cyanogen bromide agarose, about 55% of maximal activity of soluble enzyme and enzyme immobilized on amino-epoxy Sepabead derivatives, pH 9.0: about 90% of maximal activity of soluble enzyme, about 70% of maximal activity of cyanogen bromide agarose derivatives
6
9
-
pH 6: about 50% of maximal activity, pH 90: about 90% of maximal activity, hydrolysis of glutaryl-7-aminocephalosporanic acid
7
9
-
activity decreases from pH 7 to pH 9, synthesis of glutaryl-7-aminocephalosporanic acid
7
9
-
activity decrease from pH 7 to pH 9, synthesis of glutaryl-7-aminocephalosporanic acid
8
10
-
pH 8: about 50% of maximal activity, pH 9-10: maximal activity, hydrolysis of glutaryl-7-aminocephalosporanic acid
8
-
-
activity decrease from pH 7 to pH 9, synthesis of glutaryl-7-aminocephalosporanic acid
additional information
-
-
pH-dependent activity profile for the enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane is considerably expanded
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
the production of 7-aminocephalosporanic acid is carried out at 25C and pH 8.5 to avoid product degradation
34
-
-
for the 7-aminocephalosporanic acid production
37
-
-
assay at
42
-
-
enzyme immobilized on cyanogen bromide agarose
50
-
-
soluble enzyme and enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane
50
-
-
soluble enzyme and enzyme immobilized on silica gel modified by epoxide silanization
52
-
-
enzyme immobilized on amino-epoxy Sepabead derivatives
55
-
-
LX-1000EP-immobilized enzyme
55
-
-
immobilized enzyme
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
15
55
-
15C: about 40% of maximal activity, 30C: about 55% of maximal activity, 45C: about 75% of maximal activity, 55C: about 30% of maximal activity
20
60
-
20C: soluble enzyme shows 35% of maximal activity, enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane shows 45% of maximal activity, 60C: soluble enzyme shows 20% of maximal activity, enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane shows 40% of maximal activity
24
47
-
24C: about 50% of maximal activity, 47C: about 60% of maximal activity, enzyme immobilized on cyanogen bromide agarose
25
42
-
25C: about 55% of maximal activity, 48C: about 30% of maximal activity
30
55
-
30C: about 45% of maximal activity, 55C: about 70% of maximal activity
30
55
-
30C: soluble enzyme shows about 45% of maximal activity, enzyme immobilized on silica gel modified by epoxide silanization shows about 75% of maximal activity, 55C: soluble enzyme shows about 40% of maximal activity, enzyme immobilized on silica gel modified by epoxide silanization shows about 55% of maximal activity
47
57
-
47C: about 50% of maximal activity, 57C: about 60% of maximal activity, enzyme immobilized on amino-epoxy Sepabead derivatives
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
-
-
isoelectric focusing
4.6
-
-
isoelectric focusing
4.7
-
-
isoelectric focusing
5.3
-
-
isoelectric focusing, pH-range 3.5-9.5
6.5
-
-
above, isoelectric focusing
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Pseudomonas nitroreducens CCRC 11041
-
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
26000
-
-
alpha-subunit, determined by SDS-PAGE
50000
-
-
gel filtration
54000
-
-
gel filtration
58000
-
-
gel filtration
58000
-
-
SDS-PAGE
58000
-
-
beta-subunit, determined by SDS-PAGE
70000
-
-
SDS-PAGE
70000
-
O86089
SDS-PAGE
84000
-
-
SDS-PAGE
89000
-
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
1 * 70000, SDS-PAGE
?
-
1 * 26000 + 1 * 58000, SDS-PAGE
?
Brevibacillus laterosporus J1
-
1 * 70000, SDS-PAGE
-
?
Brevundimonas diminuta N176
-
1 * 26000 + 1 * 58000, SDS-PAGE
-
dimer
-
1 * 65000 + 1 * 24000
dimer
-
1 * 40000 + 1 * 22000, SDS-PAGE
dimer
-
1 * 21000 + 1 * 35000, SDS-PAGE
dimer
-
1 * 28000 + 1 * 61000, SDS-PAGE
dimer
-
1 * 58000 + 1 * 26000, SDS-PAGE
dimer
-
1 * 35000 + 1 * 21000, SDS-PAGE
dimer
-
1 * 61000 + 1* 28000, SDS-PAGE
dimer
O86089
1 * 54000 + 1 * 16000, SDS-PAGE
dimer
-
1 * 28000 + 1 * 58000, alpha- und beta-subunit, SDS-PAGE
dimer
Brevundimonas diminuta N 176
-
1 * 58000 + 1 * 26000, SDS-PAGE
-
dimer
Pseudomonas nitroreducens CCRC 11041
-
1 * 21000 + 1 * 35000, SDS-PAGE; 1 * 35000 + 1 * 21000, SDS-PAGE
-
dimer
-
1 * 61000 + 1* 28000, SDS-PAGE
-
dimer
Pseudomonas sp. BL072
-
1 * 65000 + 1 * 24000
-
dimer
Pseudomonas sp. BLO 72
-
1 * 61000 + 1* 28000, SDS-PAGE
-
dimer
-
1 * 40000 + 1 * 22000, SDS-PAGE
-
heterodimer
-
-
heterodimer
-
1 * 58000 + 1 * 28000, SDS-PAGE
heterodimer
Pseudomonas sp. N176
-
-
-
monomer
Brevibacillus laterosporus J1
-
-
-
additional information
-
the enzym is composed of 54 kDa alpha-subunits and 20 kDa beta-subunits
additional information
Brevundimonas diminuta VKM B-1297
-
the enzym is composed of 54 kDa alpha-subunits and 20 kDa beta-subunits
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
the enzyme is translated as an inactive single chain precursor, being post-translationally modified into an active enzyme. The post-translational modification takes place in two steps. The first intramolecular autocatalytic proteolysis takes place at one end of the spacer peptide by a nucleophilic Ser or Thr, which in turn becomes a new N-terminal Ser or Thr. The second intermolecular modification cleaves off the other end of the spacer peptide by another enzyme molecule
proteolytic modification
P07662
the enzyme is processed in two sequential steps of intramolecular autoproteolysis involving two distinct proteolytic mechanisms, the first mediatzed by a serine residue and the second by a glutamate
proteolytic modification
O86089
the gene encoding the enzyme is expressed as a precursor polypeptide consisting of a signal peptide followed by alpha- and beta-subunits, which are separated by a spacer peptide. Removing the signal peptide has little effect on precursor processing or enzyme activity. The precursor is supposed to be processed autocatalytically, probably intramolecularly
proteolytic modification
-
His23beta is essential for autoproteolysis, Glu455beta is responsible for the efficiency of the process
proteolytic modification
P07662
activation of precursor consists of primary and secondary autoproteolytic cleavages, generating a terminal residue with both a nucleophile and a base and releasing a nine amino acid spacer peptide. Precursor activation is likely triggered by conformational constraints within the spacer peptide, probably inducing a peptide flip. Autoproteolytic site solvent molecules, which have been trapped in a hydrophobic environment by the spacer peptide, may play a role as a general base for nucleophilic attack. The activation results in building up a catalytic triad composed of Ser170/His192/Glu624. The triad is not linked to the usual hydroxyl but the free R-amino group of the N-terminal serine residue of the native enzyme. Stabilization of a transient hydroxazolidine ring during autoproteolysis would be critical during the N to O acyl shift
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ligand bound enzyme crystals grow at 21C from hanging drops, binary complex structure of the enzyme with glutaryl-7-aminocephalosporanic acid and glutarate solved at 2.6 A and 2.5 A resolution, respectively, shows extensive interactions between the glutaryl moiety of glutaryl-7-aminocephalosporanic acid and the seven residues that form the side-chain pocket; vapour diffusion method. Binary complex structures with glutarate and glutaryl-7-aminocephalosporanic acid, solved at 2.6 A and 2.5 A resolution, respectively
Q9L5D6
hanging-drop vapour diffusion method, mutant enzymes Y202L, R226K, S170C and E159Q
P07662
polyethylene glycol 6000 as precipitant. The crystals are orthorhombic and have unit-cell parameters a = 141.41, b = 192.0, c = 80.75 A. They belong to a space group P2(1)2(1)2(1) and diffract to at least 2.7 A resolution
-
preliminary crystals of the double mutant can be grown from 4% PEG 8000, 100 mM Tris-HCl, pH 8.5, carried out by vapour diffusion using hanging-drop method at 17.85C, 1.57 A resolution
-
vapour-diffusion method. A bipyramidal crystal form is obtained from a solution containing polyethylene glycol and CaCl2. The crystal is tetragonal with the space group P4(1)2(1)2 or P4(3)2(1)2 and the unit cell parameters are a = b = 73.5 A, c = 380.3 A
-
hanging-drop vapor diffusion method using ammonium sulfate as a precipitating agent, crystal structures of the recombinant selenomethionyl native and S170A mutant precursor
P07662
the crystals of Y202L mutants including intermediates, Y202L-I, Y202-II and Y202L-II, and Q168P and L379N are grown at 22C using the hanging-drop vapor diffusion method. The three dimensional structures are determined at 2-2.5 A resolution. The pro-segments of the mutants undergo dynamic conformational changes during activation and adopt different loop conformations from one another. The autoproteolytic site is found to form catalytically competent conformation with a solvent water molecule, which is essentially conserved in the cephalosporin acylase mutants
A4ZVL3
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.5
-
-
37C, 30 min, about 75% loss of activity
4
6.5
-
37C, 30 min, stable
7
11
O86089
immobilized GLA shows good stability at pH value below 11.0
7
8
-
both forms of the enzyme, free and immobilized are partly inactivated between pH 7 and 8, while the immobilized CPC acylase shows good stability in solutions at pH 8-9.5
7
-
P07662
37C, 125 min, 10% loss of wild-type activity, 7% loss of activity of mutant enzyme K198betaA
8
9.5
-
LX-1000EP-immobilized enzyme shows good stability in solutions at pH 8.0-9.5. after 2 h of incubation
8
9.5
-
immobilized enzyme
8
-
P07662
37C, 125 min, 13% loss of wild-type activity, 7% loss of activity of mutant enzyme K198betaA
9
-
P07662
37C, 125 min, 20% loss of wild-type activity, 3% loss of activity of mutant enzyme K198betaA
10
-
P07662
37C, 125 min, 35% loss of wild-type activity, 10% loss of activity of mutant enzyme K198betaA
10
-
-
37C, 30 min, about 39% loss of activity
additional information
-
-
the stability of the enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane is significantly improved over its free form at lower pH values
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
9
-
-
the enzyme activity sharply drops on either side of pH 9
15
40
-
the LX-1000EP-immobilized enzyme is stable at temperatures up to 40C (8% loss of activity after 2 h of incubation at 40C)
25
-
-
above 25C the stability of the free enzyme decays sharply
25
-
-
there is no fatal decrease of the enzyme stability up to 25C. Above 25C, the stability of the free enzyme decays sharply
30
-
O86089
immobilized GLA shows good stability at temperature up to 30C
37
-
P07662
half-life: 68.1 h (wild-type enzyme), 88.3 h (mutant enzyme R121betaA), 107.5 h (mutant enzyme K198betaA), 53.9 h (mutant enzyme D286betA)
37
-
-
1 h, soluble enzyme and enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane are stable, t1/2 is 307.7 min for the soluble enzyme and 733.7 min for the immobilized enzyme
37
-
-
pH 8.0, t1/2: 600 min for wild-type enzyme, 51 min for beta-subunit mutant enzyme W4A, 98 min for beta-subunit mutant enzyme W4L, 12 min for beta-subunit mutant enzyme W4T, 16 min for beta-subunit mutant enzyme W4F, 307 min for beta-subunit mutant enzyme W4H, 50 min for beta-subunit mutant enzyme W4Y
37
-
-
mutant enzyme E455L is unstable at
37
-
-
60 min, soluble enzyme and enzyme immobilized on silica gel modified by epoxide silanization lose about 10% of its activity
40
-
-
immobilized enzyme, 92% activity remains, the free enzyme is nearly inactive
45
-
-
t1/2 is 49.97 min for the soluble enzyme and 200 min for the immobilized enzyme
45
-
-
pH 7.0, complete inactivation of soluble enzyme after 2 h, complete inactivation of enzyme immobilized on CNBr activated agarose and of enzyme immobilized on glyoxyl agarose, 35% inactivation of enzyme immobilized on amino-epoxy Sepabeads, 60% inactivation of enzyme immobilized on glutaraldehyde activated agarose, 70% inactivation of enzyme immobilized on epoxy Sepabeads
50
-
-
t1/2 is 29.72 min for the soluble enzyme and 65.9 min for the immobilized enzyme
50
-
-
pH 5.5, 30 min, stable up to
50
-
-
1 h, less than 10% loss of activity at pH 7.0, about 20% less of activity at pH 5, about 35% loss of activity at pH 7, about 70% loss of activity at pH 8, about 90% loss of activity at pH 9
50
-
-
1 h, stable at pH 9.0, about 45% less of activity at pH 6, about 15% loss of activity at pH 7, about 10% loss of activity at pH 8, about 60% loss of activity at pH 10
50
-
-
1 h, less than 10% loss of activity at pH 6.0, about 60% less of activity at pH 5, about 35% loss of activity at pH 7, about 50% loss of activity at pH 8, about 90% loss of activity at pH 9
50
-
-
60 min, soluble enzyme loses 77% of its activity, enzyme immobilized on silica gel modified by epoxide silanization loses 60% of its activity
55
-
-
1 h, soluble enzyme loses 77% of activity. t1/2 is 17.03 min for the soluble enzyme and 34.74 min for the immobilized enzyme
55
-
-
pH 5.5, 30 min, about 55% loss of activity
60
-
-
pH 8.0, 6 h, 25 mM Tris-HCl, 22% inactivation of wild-type enzyme, 26% inactivation of mutant enzyme M164A, 29% inactivation of mutant enzyme M164G, 20% inactivation of mutant enzyme M164L, 29% inactivation of mutant enzyme M164N, 10% inactivation of mutant enzyme M164Q
60
-
-
pH 5.5, 30 min, complete loss of activity
additional information
-
-
thermal stability of the enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane is increased significantly compared to that of the soluble enzyme
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Eupergit C-immobilized acylase has a half-life of greater than 16 days
-
FP-IDA-Ni2+ resins-immobilized GLA is recycled 21times without significant loss of activity
O86089
immobilized mutant enzyme M269Y/C305S is effective in more than 60 cycles for the production of 7-aminocephalosporanate
-
maltose-binding protein improves the solubility of the enzyme
-
the enzyme is thermostabilized by immobilization using epoxide silanization
-
the stability of the enzyme immobilized on silica gel modified with 3-aminopropyltriethoxysilane is significantly improved over its free form at lower pH values
-
in the results of repeated batch production of 7-aminocephalosporanic acid, 50% activity of the initial cycle is maintained after recycled 24 times and the average conversion rate of cephalosporin reaches 98%
-
45C, pH 7.0, complete inactivation of soluble enzyme after 2 h, complete inactivation of enzyme immobilized on CNBr activated agarose and of enzyme immobilized on glyoxyl agarose, 35% inactivation of enzyme immobilized on amino-epoxy Sepabeads, 60% inactivation of enzyme immobilized on glutaraldehyde activated agarose, 70% inactivation of enzyme immobilized on epoxy Sepabeads
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
25C, 4 weeks, mutant enzyme C305S loses 25% of its activity, mutant enzyme M269Y/C305S loses 25% of its activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, TEAE-cellulose chromatography, Toyopearl HW65F column chromatography, CM Toyopearl 650 column chromatography, Sephadex G-200 gel filtration, Q-Sepharose column chromatography, hydroxyapatite column chromatography, SP-Trisacryl gel filtration, and Sephacryl S-300gel filtration
-
ammonium sulfate precipitation, Toyopearl HW65F column chromatography, and CM Toyopearl 650 column chromatography
-
recombinant enzyme
-
a modified chitin-binding domain-GLA fusion protein is affinity purified on chitin column by changing the salt concentration of binding and elution buffer
-
ammonium sulfate precipitation, chitin granula column chromatography, and Co2+-iminodiacetic acid-Sepharose column chromatography
-
ammonium sulfate precipitation, Toyopearl HW65F column chromatography, and CM Toyopearl 650 column chromatography
-
recombinant enzyme
-
acetone precipitation and Sepharose CM column chromatography
-
by HiTrap chelating affinity chromatography
-
Cu2+-immobilized iminodiacetic acid column chromatography
-
Ni2+-HiTrap chelating column chromatography and Cu2+-immobilized iminodiacetic acid affinity chromatography
-
recombinant enzyme
-
TALON affinity resin column chromatography
O86089
overexpressed mutant protein
A4ZVL3
recombinant protein, to high purity by immobilized metal affinity chromatography, 56% recovery
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
a novel recombinant Escherichia coli strain simultaneously containing the maltose binding protein, the lysis genes and the target GLA gene in a same operon is constructed
-
expressed in Bacillus subtilis and Escherichia coli strain JM 109
-
expression in Escherichia coli
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21 (DE3) cells
-
expression in Escherichia coli
-
expressed in Escherichia coli BL21 (DE3)/pET28GA01 cells
O86089
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli BL21(DE3)/pET-Acy cells
-
expressed in Escherichia coli strain D11
-
expressed in Escherichia coli strain DH10B
-
expression in Escherichia coli BL21/DE3
P07662
expression in Escherichia coli in a two-cistron system
-
expression in Escherichia coli. Semi-large-scale fermentation of recombinant enzyme
-
expression of mutant enzymes in Escherichia coli
-
full-length wild-type enzyme or the double mutant variant H296S/H309S are transformed in Escherichia coli BL21
-
into the vector pET24 for expression in Escherichia coli BL21DE3 pLysS cells
-
expression in Escherichia coli
-
cloned in Escherichia coli BL21/DE3 using he pET23d plasmids harboring the cloned mature cephalosporin acylase gene from Pseudomonas sp. GK16 and the mutant genes E159M, D161N, D161L, L165N, Q168P, and L379N. The mutant proteins are overexpressed and purified
A4ZVL3
cloned in Escherichia coli BL21(DE3) using a pET28-acy plasmid haboring the CPC acylase gene
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D445G
-
kcat increased, turnover rate improved up to about 10fold compared to the glutaryl-7-aminocephalosporanic acid acylase activity of gamma-glutamyltranspeptidase
E423Y/E442Q/D445N
-
kcat/Km is increased, the catalytic efficiency improved up to 1000fold compared to the glutaryl-7-aminocephalosporanic acid acylase activity of gamma-glutamyltranspeptidase
D264A
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
H309A
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
S125A
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
W173A
-
kcat/KM is 2.1fold lower than wild-type value
Y126A
-
kcat/KM is 9.2 fold lower than wild-type value
Y126F
-
kcat/KM is identical to wild-type value
Y57A
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
Y57F
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
D264A
Brevibacillus laterosporus J1
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
-
H309A
Brevibacillus laterosporus J1
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
-
S125A
Brevibacillus laterosporus J1
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
-
Y57A
Brevibacillus laterosporus J1
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
-
Y57F
Brevibacillus laterosporus J1
-
no hydrolysis of glutaryl-7-aminocephalosporanic acid
-
F177H
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 17.6% of the wild-type activity, no activity with cephalosporin C
F177H
-
beta-subunit mutant, 17.6% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
F177P
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid and cephalosporin C
F177P
-
beta-subunit mutant, only partial intramolecular cleavage and no intermolecular cleavage in posttranslational modification, no activity with glutaryl-7-aminocephalosporanic acid
F177T
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 4.3% of the wild-type activity, no activity with cephalosporin C
F177T
-
beta-subunit mutant, 4.3% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate. Only partial intermolecular cleavage in posttranslational modification
L24F
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid, activity with cephalosporin C is 25.9% of the wild-type activity
L24G
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 27% of the wild-type activity, no activity with cephalosporin C
L24K
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 1.1% of the wild-type activity, no activity with cephalosporin C
L24R
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid, activity with cephalosporin C is 87.7% of the wild-type activity
L24W
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 11.3% of the wild-type activity, activity with cephalosporin C is 98.8% of wild-type activity
Q50L
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 9.9% of the wild-type activity, no activity with cephalosporin C
Q50L
-
beta-subunit mutant, 9.9% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Q50M
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 7.5% of the wild-type activity, activity with cephalosporin C is 180% of wild-type activity
Q50M
-
beta-subunit mutant, 7.5% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Q50M/F58A
-
beta-subunit mutation, activity with cephalosporin C is 180% of wild-type activity
Q50M/F58D
-
beta-subunit mutation, activity with cephalosporin C is 130% of wild-type activity
Q50M/F58G
-
beta-subunit mutation, activity with cephalosporin C is 290% of wild-type activity
Q50M/F58H
-
beta-subunit mutation, activity with cephalosporin C is 160% of wild-type activity
Q50M/F58I
-
beta-subunit mutation, activity with cephalosporin C is 150% of wild-type activity
Q50M/F58L
-
beta-subunit mutation, activity with cephalosporin C is 120% of wild-type activity
Q50M/F58P
-
beta-subunit mutation, activity with cephalosporin C is 240% of wild-type activity
Q50M/F58S
-
beta-subunit mutation, activity with cephalosporin C is 260% of wild-type activity
Q50M/F58T
-
beta-subunit mutation, activity with cephalosporin C is 230% of wild-type activity
Q50M/Y149F
-
Q50M mutation in beta-subunit, Y149F mutation in alpha-subunit, activity with cephalosporin C is 157% of wild-type activity
Q50M/Y149K
-
Q50M mutation in beta-subunit, Y149K mutation in alpha-subunit, activity with cephalosporin C is 600% of wild-type activity
Q50M/Y149K/F177G
-
beta-subunit mutation, activity with cephalosporin C is 787% of wild-type activity
Q50M/Y149K/F177P
-
beta-subunit mutation, activity with cephalosporin C is 301% of wild-type activity
Q50M/Y149K/F177S
-
beta-subunit mutation, activity with cephalosporin C is 456% of wild-type activity
Q50M/Y149K/M145A
-
Q50M and Y149K are mutation in the beta-subunit, M145A is a mutation in the alpha-subunit, activity with cephalosporin C is 335% of wild-type activity
Q50M/Y149K/M145L
-
Q50M and Y149K are mutations in the beta-subunit, M145L is a mutation in the alpha-subunit, activity with cephalosporin C is 704% of wild-type activity
Q50M/Y149K/M145P
-
Q50M and Y149K are mutations in the beta-subunit, M145P is a mutation in the alpha-subunit, activity with cephalosporin C is 442% of wild-type activity
Q50M/Y149K/M145T
-
Q50M and Y149K are mutation in the beta-subunit, M145T is a mutation in the alpha-subunit, activity with cephalosporin C is 433% of wild-type activity
Q50M/Y149K/Y33N
-
beta-subunit mutation, activity with cephalosporin C is 460% of wild-type activity
Q50M/Y149K/Y33S
-
beta-subunit mutation, activity with cephalosporin C is 568% of wild-type activity
Q50M/Y149K/Y33T
-
beta-subunit mutation, activity with cephalosporin C is 424% of wild-type activity
Q50M/Y33D
-
beta-subunit mutant, activity with cephalosporin C is 450% of wild-type activity
Q50M/Y33G
-
beta-subunit mutant, activity with cephalosporin C is 70% of wild-type activity
Q50M/Y33H
-
beta-subunit mutant, activity with cephalosporin C is 101% of wild-type activity
Q50M/Y33L
-
beta-subunit mutant, activity with cephalosporin C is 202% of wild-type activity
Q50M/Y33N
-
beta-subunit mutant, activity with cephalosporin C is 350% of wild-type activity
Q50M/Y33P
-
beta-subunit mutant, activity with cephalosporin C is 99% of wild-type activity
Q50M/Y33S
-
beta-subunit mutant, activity with cephalosporin C is 60% of wild-type activity
Q50M/Y33T
-
beta-subunit mutant, activity with cephalosporin C is 109% of wild-type activity
Q50M/Y33V
-
beta-subunit mutant, activity with cephalosporin C is 135% of wild-type activity
Q50R
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 2.1% of the wild-type activity, no activity with cephalosporin C
Q50R
-
beta-subunit mutant, 2.1% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Q50T
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 72.5% of the wild-type activity, no activity with cephalosporin C
Q50T
-
beta-subunit mutant, 72.5% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Q50Y
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 15.7% of the wild-type activity, no activity with cephalosporin C
Q50Y
-
beta-subunit mutant, 15.7% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
R155G
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 88% of the wild-type activity
R57C
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid and cephalosporin C
R57C
-
beta-subunit mutant, no intermolecular cleavage in posttranslational modification, no activity with glutaryl-7-aminocephalosporanic acid
R57I
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid and cephalosporin C
R57I
-
beta-subunit mutant, no intermolecular cleavage in posttranslational modification, no activity with glutaryl-7-aminocephalosporanic acid
R57K
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 0.3% of the wild-type activity, no activity with cephalosporin C
R57K
-
beta-subunit mutant, 0.3% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, only partial intermolecular cleavage in posttranslational modification
R57S
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid and cephalosporin C
S152G
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 75.4% of the wild-type activity, activity with cephalosporin C is 38.6% of wild-type activity
S152H
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 63.7% of the wild-type activity, activity with cephalosporin C is 11.9% of wild-type activity
S152N
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 78% of the wild-type activity, activity with cephalosporin C is 24.8% of wild-type activity
S152P
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 76.2% of the wild-type activity, activity with cephalosporin C is 60.4% of wild-type activity
S152T
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 81.4% of the wild-type activity, activity with cephalosporin C is 34.7% of wild-type activity
S1A
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid and cephalosporin C
S1A
-
beta-subunit mutant, no processing takes place, no activity with glutaryl-7-aminocephalosporanic acid
S1betaC
-
no activity with glutaryl-7-aminocephalosporanic acid and cephalosporin C
S1C
-
beta-subunit mutant, no intermolecular cleavage in posttranslational modification, no activity with glutaryl-7-aminocephalosporanic acid
V70G
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid, activity with cephalosporin C is 32.1% of the wild-type activity
V70S
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 28.8% of the wild-type activity, no activity with cephalosporin C
Y149C
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 16.4% of the wild-type activity, activity with cephalosporin C is 24% of wild-type activity
Y149C
-
alpha-subunit mutant, 16.4% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Y149G
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 0.7% of the wild-type activity, no activity with cephalosporin C
Y149G
-
alpha-subunit mutant, 0.7% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Y149L
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 6.3% of the wild-type activity, activity with cephalosporin C is 48% of wild-type activity
Y149L
-
alpha-subunit mutant, 6.3% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Y149N
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 30.8% of the wild-type activity, activity with cephalosporin C is 48% of wild-type activity
Y149N
-
alpha-subunit mutant, 30.8% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Y149P
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 1.1% of the wild-type activity, no activity with cephalosporin C
Y149P
-
alpha-subunit mutant, 1.1% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Y149R
-
alpha-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 13.6% of the wild-type activity, no activity with cephalosporin C
Y149R
-
alpha-subunit mutant, 13.6% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Y33F
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 65.4% of the wild-type activity, no activity with cephalosporin C
Y33F
-
beta-subunit mutant, 65.4% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
Y33I
-
beta-subunit mutant, activity with glutaryl-7-aminocephalosporanic acid is 9.3% of the wild-type activity, activity with cephalosporin C is 90% of wild-type activity
Y33I
-
beta-subunit mutant, 9.3% of the wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate only partial intermolecular cleavage in posttranslational modification
Y33S
-
beta-subunit mutant, no activity with glutaryl-7-aminocephalosporanic acid, activity with cephalosporin C is 120% of the wild-type activity
Y33S
-
beta-subunit mutant, no intermolecular cleavage in posttranslational modification, no activity with glutaryl-7-aminocephalosporanic acid
A215E
-
Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 7.6fold lower than wild-type value
A215F
-
Vmax/Km for cephalosporin C is 2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 3.9fold lower than wild-type value
A215L
-
Vmax/Km for cephalosporin C is 1.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 4.2fold lower than wild-type value
A215V
-
Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.5fold lower than wild-type value
A215Y
-
Vmax/Km for cephalosporin C is 4.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.6fold lower than wild-type value
A215Y/H296S
-
Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 25.1fold lower than wild-type value
A215Y/H296S/H309S
-
Vmax/Km for cephalosporin C is 3.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 13.7fold lower than wild-type value
A215Y/H309S
-
Vmax/Km for cephalosporin C is 3.8fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 2.9fold higher than wild-type value
A271F
-
118% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 56.2% of wild-type activity with cephalosporin C as substrate
A271L
-
104% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 100% of wild-type activity with cephalosporin C as substrate
A271Y
-
101% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 122% of wild-type activity with cephalosporin C as substrate
A675G
-
the mutant exhibits 112.8% activity with cephalosporin C compared to the wild type enzyme; the mutant exhibits significantly enhanced specific enzymatic activity compared to the wild type enzyme (35% increase)
C102S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
C199S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
C277S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
C305S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C. Expression in Escherichia coli is 2-3fold higher than that of the wild-type enzyme
C391S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
C493S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
C496S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
C748S
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
D286betaA
P07662
beta-subunit mutant enzyme, KM-value is 1.1fold lower than the wild-type value, turnover-number is 1.03fold higher than the wild-type value. Half-life at 37C is 53.9 h compared to 68.1 h for wild-type enzyme. Optimal pH is pH 9.0 compared to pH 7.0 for wild-type enzyme
D416Y
-
Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.2fold lower than wild-type value
D416Y/H417Y
-
Vmax/Km for cephalosporin C is 5.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 151fold lower than wild-type value
E159D
-
the mutant is still active, but significantly retarded in the second autocleavage compared to the wild type enzyme
E89A/A215Y
-
Vmax/Km for cephalosporin C is 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 25.2fold lower than wild-type value
F229L
P07662
turnover number for adipyl-7-aminodesacetoxycephalosporanic acid is nealy identical to wild-type value, 1.8fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.4fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 1.45fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme
F270M
-
Vmax/Km for cephalosporin C is 1.8fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.7fold lower than wild-type value
F375L
P07662
1.5fold increase in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 1.2fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 3.9fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 8.2fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme. Improved activity ratio for adipyl-7-aminodesacetoxycephalosporanic acid to glutaryl-7-aminocephalosporanic acid of the mutant enzyme is a consequence of a decreased catalytic efficiency towards glutaryl-7-aminocephalosporanic acid
F375L
-
decreased activity compared to the wild type enzyme
F375M
-
decreased activity compared to the wild type enzyme
G160A
-
the mutant performs the second autocleavage more slowly than the wild type enzyme
G160L
-
the mutant performs the second autocleavage more slowly than the wild type enzyme
H296F
-
Vmax/Km for cephalosporin C is 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 50.3fold lower than wild-type value
H296N
-
Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is fold lower than wild-type value
H296S
-
Vmax/Km for cephalosporin C is 1.7fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 10.1fold lower than wild-type value
H296S/H309S
-
Vmax/Km for cephalosporin C is 4fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 21.6fold lower than wild-type value
H296S/H309S
-
the mutant also shows activity with cephalosporin C (3fold increase compared to the wild type enzyme)
H296S/H417M
-
Vmax/Km for cephalosporin C is 2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 151fold lower than wild-type value
H296T
-
Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.2fold lower than wild-type value
H309S
-
Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 8.9fold lower than wild-type value
H309V
-
the mutant exhibits 88.9% activity with cephalosporin C compared to the wild type enzyme
H309Y
-
Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 16.8fold lower than wild-type value
H417Y
-
Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 4.4fold lower than wild-type value
I44V/E49stop/D416Y/H417Y
-
Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is more than 1510fold lower than wild-type value
K100Q
-
81% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 106% of wild-type activity with cephalosporin C as substrate
K114Q
-
86% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 101% of wild-type activity with cephalosporin C as substrate
K170Q
-
130% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 95.6% of wild-type activity with cephalosporin C as substrate
K187Q
-
113% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 91.1% of wild-type activity with cephalosporin C as substrate
K198betaA
P07662
beta-subunit mutant enzyme, KM-value is 1.04fold higher than the wild-type value, turnover-number is 1.1fold lower than the wild-type value. Half-life at 37C is 107.5 h compared to 68.1 h for wild-type enzyme. Optimal pH is pH 8.0 compared to pH 7.0 for wild-type enzyme. Mutant enzyme shows higher stability at alkaline pH than wild-type enzyme
K255Q
-
107% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 97% of wild-type activity with cephalosporin C as substrate
K301Q
-
101% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
K44Q
-
102% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 111% of wild-type activity with cephalosporin C as substrate
K507Q
-
102% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 113.9% of wild-type activity with cephalosporin C as substrate
K629Q
-
94.2% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate
K73Q
-
46.9% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 47% of wild-type activity with cephalosporin C as substrate
L666F
-
the mutant exhibits significantly reduced specific enzymatic activity (75.4%) with cephalosporin C compared to the wild type enzyme
L677A
-
the mutant exhibits significantly reduced specific enzymatic activity compared to the wild type enzyme
M116A
-
at pH 7.5, 76.9% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 95.8% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 108% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate
M157A
-
at pH 7.5, 70.9% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 58% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 65% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is lower than that of wild-type enzyme
M164A
-
at pH 7.5, 167% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 104% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 86.6% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is higher than that of wild-type enzyme. The ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.2fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 2.16fold higher than the wild-type ratio
M164F
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.3fold lower than the wild-type ratio
M164G
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.3fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 2.23fold higher than the wild-type ratio
M164L
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 2.1fold lower than the wild-type ratio
M164N
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.4fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 1.81fold higher than the wild-type ratio
M164P
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.3fold lower than the wild-type ratio
M164Q
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 5.3fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is nearly identical to the wild-type ratio
M164S
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.5fold lower than the wild-type ratio
M164T
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.4fold lower than the wild-type ratio
M174A
-
at pH 7.5, 96.5% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 109% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 122% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate
M227A
-
at pH 7.5, 106% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 74.2% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 105% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is lower than that of wild-type enzyme
M269F
-
98.2% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 165% of wild-type activity with cephalosporin C as substrate. The ratio of turnover number to Km-value with cephalosporin C as substrate is 1.8fold higher than the wild-type ratio
M269L
-
92.5% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 107.9% of wild-type activity with cephalosporin C as substrate
M269Y
-
91.5% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 155% of wild-type activity with cephalosporin C as substrate. The ratio of turnover number to Km-value with cephalosporin C as substrate is 1.6fold higher than the wild-type ratio
M269Y/C305S
-
1.6fold higher activity with cephalosporin C than wild-type enzyme
M271V/Q291K/T374S
P07662
1.3fold increase in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 1.4fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.2fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 1.3fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme
M98A
-
at pH 7.5, 126% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 83.1% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 91.2% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate
N266H
P07662
1.2fold increase in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 6.9fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.3fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 2fold decrease in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme. Nearly 10fold improved catalytic efficiency on adipyl-7-aminodesacetoxycephalosporanic acid, resulting from a 50% increase in turnover-number and a 6fold decrease in KM-value without decreasing the catalytic efficiency on glutaryl-7-aminocephalosporanic acid
N266H
-
decreased activity compared to the wild type enzyme
N266M
-
decreased activity compared to the wild type enzyme
N266Q
-
decreased activity compared to the wild type enzyme
N266Q/F375L
-
decreased activity compared to the wild type enzyme
N266S
P07662
1.4fold decrease in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 2.3fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.95fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 2.8fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme
P295A
-
the mutant exhibits 20% activity with cephalosporin C compared to the wild type enzyme
Q50betaN/K198betaA
P07662
beta-subunit mutant enzyme, KM-value is 2.14fold lower than the wild-type value, turnover-number is 1.45fold higher than the wild-type value, immobilized mutant enzyme shows 34.2% increase in specific activity compared to immobilized wild-type enzyme. Ability of the mutant enzyme to hydrolyze adipoyl 6-aminopenicillinic acid is improved, compared to activity of wild-type enzyme
Q50N
P07662
beta-subunit mutant enzyme, KM-value is 2.05fold lower than the wild-type value, turnover-number is 1.45fold higher than the wild-type value. Ability of the mutant enzyme to hydrolyze adipoyl 6-aminopenicillinic acid is improved, compared to activity of wild-type enzyme
R121A
P07662
beta-subunit mutant enzyme, KM-value is identical to the wild-type value, turnover-number is nearly identical to wild-type value. Half-life at 37C is 88.3 h compared to 68.1 h for wild-type enzyme. Optimal pH is pH 6.0 compared to pH 7.0 for wild-type enzyme
S170C
-
the mutant simultaneously loses the activities for both the second autocleavage and substrate hydrolysis
S22P/T394P/D416Y/H417Y
-
Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is more than 1510fold lower than wild-type value
S293C
-
1.21% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, no activity with cephalosporin C as substrate
Y151F
P07662
alpha-subunit mutant enzyme, KM-value is 1.9fold lower than the wild-type value, turnover-number is 1.17fold higher than the wild-type value. Ability of the mutant enzyme to hydrolyze adipoyl 6-aminopenicillinic acid is improved, compared to activity of wild-type enzyme
Y151F/Q50N
P07662
mutant enzyme with mutation Y151F in alpha-subunit and mutation Q50N in beta-subunit, KM-value is 1.2fold lower than the wild-type value, turnover-number is 1.09fold higher than the wild-type value
Y178F/F375H
-
the mutation synergistically improve the catalytic efficiency towards adipyl-7-ADCA 36fold, The activity of this double mutant towards adipyl-7-ADCA is 50% of the activity of the wild-type enzyme towards the preferred substrate glutaryl-7-aminocephalosporanic acid
Y178H
-
decreased activity compared to the wild type enzyme
Y178H/N266H
-
decreased activity compared to the wild type enzyme
Y178H/N266M
-
decreased activity compared to the wild type enzyme
Y178H/N266Q/F375L
-
decreased activity compared to the wild type enzyme
Y215Y/F270S
-
Vmax/Km for cephalosporin C is 2.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.2fold lower than wild-type value
Y270A
-
70.6% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 24.3% of wild-type activity with cephalosporin C as substrate
Y270E
-
no activity with glutaryl-7-aminocephalosporanic acid or cephalosporin C as substrate
Y270F
-
50.4% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 46.1% of wild-type activity with cephalosporin C as substrate. The ratio of turnover number to Km-value with cephalosporin C as substrate is 2.6fold lower than the wild-type ratio
Y270L
-
28.1% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 32.3% of wild-type activity with cephalosporin C as substrate
Y270S
-
61.7% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 28.3% of wild-type activity with cephalosporin C as substrate
Y271F
-
Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.4fold lower than wild-type value
Y27V
-
Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 2.1fold lower than wild-type value
E455D
-
the ratio of turnover number to Km-value of the beta-subunit mutant enzyme is 4.5fold lower than the wild-type ratio
E455L
-
the ratio of turnover number to Km-value of the beta-subunit mutant enzyme is 34fold lower than the wild-type ratio
E455Q
-
the ratio of turnover number to Km-value of the beta-subunit mutant enzyme is 6.1fold lower than the wild-type ratio
H23D
-
the ratio of turnover number to Km-value of the beta-subunit mutant enzyme is 9.7fold lower than to the wild-type ratio
W4A
-
beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.6fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 2.2fold lower than wild-type value, t1/2 at 37C is 9.8fold lower than wild-type value
W4F
-
beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.8fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 1.6fold lower than wild-type value, t1/2 at 37C is 37.5fold lower than wild-type value
W4H
-
beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.9fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 1.4fold lower than wild-type value, t1/2 at 37C is 1.9fold lower than wild-type value
W4L
-
beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.8fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 2.1fold lower than wild-type value, t1/2 at 37C is 6.1fold lower than wild-type value
W4T
-
beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.7fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 2.4fold lower than wild-type value, t1/2 at 37C is 50fold lower than wild-type value
W4Y
-
beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.6fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 1.3fold lower than wild-type value, t1/2 at 37C is 12fold lower than wild-type value
A215Y
Pseudomonas sp. N176
-
Vmax/Km for cephalosporin C is 4.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.6fold lower than wild-type value
-
C102S
Pseudomonas sp. N176
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
-
C199S
Pseudomonas sp. N176
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
-
C277S
Pseudomonas sp. N176
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
-
C305S
Pseudomonas sp. N176
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C. Expression in Escherichia coli is 2-3fold higher than that of the wild-type enzyme
-
C391S
Pseudomonas sp. N176
-
mutant enzyme retains activity towards glutaryl-7-aminocephalosporanic acid and cephalosporin C
-
F270M
Pseudomonas sp. N176
-
Vmax/Km for cephalosporin C is 1.8fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.7fold lower than wild-type value
-
H296S
Pseudomonas sp. N176
-
Vmax/Km for cephalosporin C is 1.7fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 10.1fold lower than wild-type value
-
H309S
Pseudomonas sp. N176
-
Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 8.9fold lower than wild-type value
-
K100Q
Pseudomonas sp. N176
-
81% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 106% of wild-type activity with cephalosporin C as substrate
-
K114Q
Pseudomonas sp. N176
-
86% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 101% of wild-type activity with cephalosporin C as substrate
-
K255Q
Pseudomonas sp. N176
-
107% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 97% of wild-type activity with cephalosporin C as substrate
-
K44Q
Pseudomonas sp. N176
-
102% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 111% of wild-type activity with cephalosporin C as substrate
-
K73Q
Pseudomonas sp. N176
-
46.9% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 47% of wild-type activity with cephalosporin C as substrate
-
M116A
Pseudomonas sp. N176
-
at pH 7.5, 76.9% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 95.8% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 108% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate
-
M157A
Pseudomonas sp. N176
-
at pH 7.5, 70.9% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 58% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 65% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is lower than that of wild-type enzyme
-
M164A
Pseudomonas sp. N176
-
at pH 7.5, 167% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 104% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 86.6% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is higher than that of wild-type enzyme. The ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.2fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 2.16fold higher than the wild-type ratio
-
M164G
Pseudomonas sp. N176
-
the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.3fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 2.23fold higher than the wild-type ratio
-
M98A
Pseudomonas sp. N176
-
at pH 7.5, 126% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 83.1% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 91.2% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate
-
Y271F
Pseudomonas sp. N176
-
Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.4fold lower than wild-type value
-
F375L
Pseudomonas sp. SY-77
-
decreased activity compared to the wild type enzyme
-
F375M
Pseudomonas sp. SY-77
-
decreased activity compared to the wild type enzyme
-
N266H
Pseudomonas sp. SY-77
-
decreased activity compared to the wild type enzyme
-
N266M
Pseudomonas sp. SY-77
-
decreased activity compared to the wild type enzyme
-
N266Q
Pseudomonas sp. SY-77
-
decreased activity compared to the wild type enzyme
-
F375A
-
turnover number is 2.45fold lower than wild-type value, KM-value is 1.9fold lower than wild-type value, turnover number is 1.7fold lower than wild-type value, KM-value is 2.2fold higher than wild-type value
F375C
-
turnover number is 1.1fold higher than wild-type value, KM-value is 5.9fold lower than wild-type value, turnover number is 1.7fold lower than wild-type value, KM-value is 1.2fold higher than wild-type value
F375D
-
turnover number is 32.7fold lower than wild-type value, KM-value is 11fold higher than wild-type value, turnover number is 400fold lower than wild-type value, KM-value is 7fold higher than wild-type value
F375E
-
turnover number is 6.1fold lower than wild-type value, KM-value is 12fold higher than wild-type value, turnover number is 500fold lower than wild-type value, KM-value is 6.7fold higher than wild-type value
F375G
-
turnover number is 6.7fold lower than wild-type value, KM-value is 6.25fold lower than wild-type value, turnover number is 1.6fold lower than wild-type value, KM-value is 2.1fold higher than wild-type value
F375H
-
turnover number is 2.4fold higher than wild-type value, KM-value is 1.1fold lower than wild-type value, turnover number is 3fold lower than wild-type value, KM-value is 3.5fold higher than wild-type value
F375I
-
turnover number is 5fold lower than wild-type value, KM-value is 2.1fold higher than wild-type value, turnover number is 6fold lower than wild-type value, KM-value is 3.2fold higher than wild-type value
F375K
-
turnover number is 13.6fold lower than wild-type value, KM-value is 2.2fold higher than wild-type value, turnover number is 1.8fold lower than wild-type value, KM-value is 22.6fold higher than wild-type value
F375L
-
turnover number is 1.4fold higher than wild-type value, KM-value is 1.4fold lower than wild-type value, turnover number is 3.6fold lower than wild-type value, KM-value is 8.4fold higher than wild-type value
F375M
-
turnover number is 1.3fold higher than wild-type value, KM-value is 1.5fold lower than wild-type value, turnover number is 2fold lower than wild-type value, KM-value is 5.5fold higher than wild-type value
F375N
-
turnover number is 1.8fold higher than wild-type value, KM-value is 1.2fold lower than wild-type value, turnover number is 2fold lower than wild-type value, KM-value is 5.8fold higher than wild-type value
F375Q
-
turnover number is 1.4fold higher than wild-type value, KM-value is 2.1fold higher than wild-type value, turnover number is 5fold lower than wild-type value, KM-value is 22.6fold higher than wild-type value
F375R
-
turnover number is 27.2fold lower than wild-type value, KM-value is 10fold higher than wild-type value, turnover number is 19fold lower than wild-type value, KM-value is 19fold higher than wild-type value
F375S
-
turnover number is 4.5fold lower than wild-type value, KM-value is nearly identical to wild-type value, turnover number is 1.3fold lower than wild-type value, KM-value is 2.3fold higher than wild-type value
F375T
-
turnover number is 2fold lower than wild-type value, KM-value is 1.1fold lower than wild-type value, turnover number is 2.2fold lower than wild-type value, KM-value is 2.9fold higher than wild-type value
F375V
-
turnover number is 5.4fold lower than wild-type value, KM-value is 1.4fold higher than wild-type value, turnover number is 3.9fold lower than wild-type value, KM-value is 3.2fold higher than wild-type value
F375W
-
turnover number is 18.8fold lower than wild-type value, KM-value is 1.5fold higher than wild-type value, turnover number is 8.5fold lower than wild-type value, KM-value is 25.8fold higher than wild-type value
F375Y
-
turnover number is 1.14fold lower than wild-type value, KM-value is 2.4fold lower than wild-type value, turnover number is 1.1fold lower than wild-type value, KM-value is 2.6fold higher than wild-type value
N266A
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 2.3fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2.9fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 8.4fold higher than wild-type value
N266C
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.3fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.5fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 5.2fold higher than wild-type value
N266D
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 205fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold higher than wild-type value. Less efficient cleavage of the spacer from the alpha-subunit compared to wild-type enzyme
N266E
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 17.8fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 11.5fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 160fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 9.4fold higher than wild-type value. Less efficient cleavage of the spacer from the alpha-subunit compared to wild-type enzyme
N266F
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.1fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 4.4fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 3.2fold higher than wild-type value
N266G
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 17.4fold higher than wild-type value
N266H
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.15fold higher than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 8.6fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 1.3fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 1.4fold higher than wild-type value. 100% improved conversion of cephalosporin C compared to wild type enzyme
N266L
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.64fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 48.8fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 13.9fold higher than wild-type value
N266M
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 4.6fold higher than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 3.6fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2.2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 5.4fold higher than wild-type value
N266P
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 6.7fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 3.7fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 30.8fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 41.9fold higher than wild-type value
N266Q
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.12fold higher than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.9fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 1.7fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 3.9fold higher wild-type value. 100% improved conversion of cephalosporin C compared to wild type enzyme
N266S
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.7fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2.5fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 2.4fold higher than wild-type value
N266T
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 2.7fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 4.75fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 8.2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 48.4fold higher than wild-type value
N266W
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 3fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 5.8fold higher than wild-type value. 30% improved conversion of cephalosporin C compared to wild type enzyme
N266Y
-
turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 6.6fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 3.5fold higher than wild-type value
H23Q
-
the ratio of turnover number to Km-value of the beta-subunit mutant enzyme is 4.3fold lower than the wild-type ratio
additional information
O86089
substitution of the first residue of the beta-subunit, Ser, results in a complete loss of enzyme activity, and substitution of the last residue of the spacer, Gly, leads to an inactive and unprocessed precursor
additional information
-
deletion of the signal peptide and mutation in the alpha-subunit of the acylase has little influence on its posttranslational processing and its kinetic parameters
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
industry
-
great potential for industrial application of the lysis genes-assisted cell disruption
industry
Achromobacter xylosoxidans MTCC *491
-
great potential for industrial application of the lysis genes-assisted cell disruption
-
synthesis
-
D-amino acid oxidase in the permeabilized Pichia pastoris cells and immobilized glutaryl-7-aminocephalosporanic acid acid acylase on support, are employed to convert cephalosporin to 7-aminocephalosporanic acid in a single reactor
biotechnology
-
with an aim to increase the yield of 7-aminocephalosporanic acid production, a stepwise strategy, statistical medium is applied for optimizing the medium composition for the production of CPC acylase
biotechnology
-
a biological and colometric method is evaluated for determination of cephalosporin acylase product in bacteria
industry
-
the huge production of engineered GLA produced in Escherichia coli under optimized conditions makes this enzyme an economic tool to be used in bioconversion processes at the industrial level, e.g., in 7-ACA production
pharmacology
-
the product of the reaction 7-aminocephalosporanic acid is a starting material for semisynthetic cephalosporin antibiotics. High thermal stability of the enzyme immobilized on silica gels indicates that it can be successfully used for the production of 7-aminocephalosporanic acid on an industrial scale
synthesis
-
immobilized mutant enzyme M269Y/C305S may be a promising enzyme in one-step enzymatic production of 7-aminocephalosporanate from cephalosporin C
synthesis
P07662
enzymatic production of 7-aminocephalosporanate. Construction of site-directed mutants with enhanced activity and stability
synthesis
-
mutant enzyme M269Y may be a promising enzyme in the one-step enzymic production of 7-amino-cephalosporanic acid from cephalosporin C
synthesis
-
bioconversion of cephalosporin C into 7-aminocephalosporanate
synthesis
-
production of 7-aminocephalosporanate on a large scale at 25C using enzyme immobilized by epoxide silanization
pharmacology
Pseudomonas sp. KAC-1
-
the product of the reaction 7-aminocephalosporanic acid is a starting material for semisynthetic cephalosporin antibiotics. High thermal stability of the enzyme immobilized on silica gels indicates that it can be successfully used for the production of 7-aminocephalosporanic acid on an industrial scale
-
synthesis
Pseudomonas sp. KAC-1
-
production of 7-aminocephalosporanate on a large scale at 25C using enzyme immobilized by epoxide silanization
-
industry
Pseudomonas sp. N176
-
the huge production of engineered GLA produced in Escherichia coli under optimized conditions makes this enzyme an economic tool to be used in bioconversion processes at the industrial level, e.g., in 7-ACA production
-
synthesis
Pseudomonas sp. N176
-
bioconversion of cephalosporin C into 7-aminocephalosporanate; immobilized mutant enzyme M269Y/C305S may be a promising enzyme in one-step enzymatic production of 7-aminocephalosporanate from cephalosporin C; mutant enzyme M269Y may be a promising enzyme in the one-step enzymic production of 7-amino-cephalosporanic acid from cephalosporin C
-
synthesis
-
immobilzation on LX-1000EP, an epoxy support, with a recovery of 33.8% and an activity of 81 U/g wet resin, suggesting a potential in industrial 7-aminocephalosporanic acid production. Under optimal conditions, a very high 7-aminocephalosporanic acid yield of 96.6% is obtained within 60 min