Information on EC 3.5.1.19 - nicotinamidase

Word Map on EC 3.5.1.19
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
3.5.1.19
-
RECOMMENDED NAME
GeneOntology No.
nicotinamidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
nicotinamide + H2O = nicotinate + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
aldoxime degradation
-
-
Metabolic pathways
-
-
NAD salvage pathway I
-
-
Nicotinate and nicotinamide metabolism
-
-
pyridine nucleotide cycling (plants)
-
-
NAD metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
nicotinamide amidohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9033-32-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
strain 544, ATCC 23448, gene pncA, a facultative intracellular pathogen
-
-
Manually annotated by BRENDA team
strain 544, ATCC 23448, gene pncA, a facultative intracellular pathogen
-
-
Manually annotated by BRENDA team
K12
-
-
Manually annotated by BRENDA team
Flavobacterium peregrinum
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
i.e. Micrococcus lysodeiktikus
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
deep-sea extremely halotolerant and alkaliphilic organism
UniProt
Manually annotated by BRENDA team
deep-sea extremely halotolerant and alkaliphilic organism
UniProt
Manually annotated by BRENDA team
strain GDI 211
-
-
Manually annotated by BRENDA team
strain GDI 211
-
-
Manually annotated by BRENDA team
strain CA401, strain FA18, strain FA64, strain FA86, strain FA131, strain 569B
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
-
nicotinamidases are a family of peptide hydrolases that generally contain a Zn2+ ion
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-methylnicotinamide + H2O
1-methylnicotinate + NH3
show the reaction diagram
-
-
-
-
?
4-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methyl nicotinamide + H2O
5-methyl nicotinate + NH3
show the reaction diagram
5-methylnicotinamide + H2O
5-methylnicotinate + NH3
show the reaction diagram
5-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
6-aminopyridine-3-carboxamide + H2O
?
show the reaction diagram
6-chloropyridine-3-carboxamide + H2O
?
show the reaction diagram
6-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
benzamide + H2O
benzoate + NH3
show the reaction diagram
-
-
-
-
?
ethyl nicotinate + H2O
nicotinate + ethanol
show the reaction diagram
-
-
-
?
ethyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
methyl nicotinate + H2O
nicotinate + methanol
show the reaction diagram
-
-
-
?
methyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
nicotinamide + H2O
NH3 + nicotinic acid
show the reaction diagram
nicotinamide + H2O
nicotinate + NH3
show the reaction diagram
nicotinamide + H2O
nicotinic acid + NH3
show the reaction diagram
p-nitrophenol + H2O
phenol + NH3
show the reaction diagram
-
-
-
-
?
p-nitrophenyl nicotinate + H2O
?
show the reaction diagram
-
-
-
-
?
phenyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
pyrazinamide + H2O
NH3 + pyrazinoic acid
show the reaction diagram
pyrazinamide + H2O
pyrazine-2-carboxylic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinoic acid + NH3
show the reaction diagram
pyridine-3-carbothioamide + H2O
?
show the reaction diagram
-
44% activity compared to nicotinamide
-
-
?
thionicotinamide + H2O
pyridine-3-carbothioic O-acid
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methoxypyridine-3-carboxamide + H2O
?
show the reaction diagram
5-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
6-aminopyridine-3-carboxamide + H2O
?
show the reaction diagram
6-chloropyridine-3-carboxamide + H2O
?
show the reaction diagram
6-methylpyridine-3-carboxamide + H2O
?
show the reaction diagram
ethyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
methyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
nicotinamide + H2O
NH3 + nicotinic acid
show the reaction diagram
nicotinamide + H2O
nicotinate + NH3
show the reaction diagram
nicotinamide + H2O
nicotinic acid + NH3
show the reaction diagram
phenyl pyridine-3-carboxylate + H2O
?
show the reaction diagram
pyrazinamide + H2O
NH3 + pyrazinoic acid
show the reaction diagram
pyrazinamide + H2O
pyrazine-2-carboxylic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinic acid + NH3
show the reaction diagram
pyrazinamide + H2O
pyrazinoic acid + NH3
show the reaction diagram
-
a nicotinamide prodrug. In Mycobacterium tuberculosis the enzyme converts the nicotinamide analogue prodrug pyrazinamide into the bacteriostatic pyrazinoic acid, hence the alternative name, pyrazinamidase, PncA
a bacteriostatic drug
-
?
pyridine-3-carbothioamide + H2O
?
show the reaction diagram
-
44% activity compared to nicotinamide
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activation
Ni2+
-
enhances activity
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl
-
Mg2+ reverses inhibition
2-chloropyridine-3-carbaldehyde
2-mercaptoethanol
-
-
3-(3-pyridyl)-1-propanol
-
moderate
3-acetylpyridine
3-aminomethylpyridine
-
potent
3-chloropyridine
-
moderate
3-Cyanopyridine
3-hydroxypyridine
-
-
3-indolepropionic acid
-
inhibits amidase and esterase activity
3-pyridine carboxaldehyde
-
competitive inhibitor
3-Pyridineacetonitrile
-
moderate
3-pyridinealdoxime
-
potent
3-Pyridinecarboxaldehyde
-
potent
3-Pyridylcarbinol
-
potent
4-aminonicotinamide
-
potent
4-methoxylnicotinaldehyde
5-bromonicotinaldehyde
competitive inhibition
-
5-bromopyridine-3-carbaldehyde
5-methoxynicotinaldehyde
Q97PM2
competitive inhibitor
5-methoxypyridine-3-carbaldehyde
5-methylnicotinaldehyde
5-Methylnicotinamide
5-methylpyridine-3-carbaldehyde
6-fluoropyridine-3-carbaldehyde
8-hydroxyquinoline
-
-
benzaldehyde
-
competitive inhibition
Benzoic acid
-
inhibits amidase and esterase activity
Carbobenzoxyamido-2-phenyl-ethyl-chloromethyl-ketone
-
-
diisopropylfluorophosphate
-
-
dithiothreitol
-
-
Fe3+
1 mM, 94% inhibition
guanidinium hydrochloride
-
3.4 M, complete inhibition
HgCl2
Flavobacterium peregrinum
-
-
Hydrocinnamic acid
-
inhibits amidase and esterase activity
iodoacetamide
-
reversible, competitive inhibitor
Isoniazid
-
moderate
L-thyroxine
-
-
N,N'-diethylnicotinamide
Flavobacterium peregrinum
-
competitive
N-ethylmaleimide
-
-
N2-ethylnicotinamide
Flavobacterium peregrinum
-
competitive
N2-methylnicotinamide
Flavobacterium peregrinum
-
competitive
NAD+
-
competitive with respect to nicotinamide
nicotinal hydroxamic acid
-
moderate
nicotinaldehyde
nicotinamide
-
inhibits esterolysis of p-nitrophenyl acetate
nicotinamide analogs
Flavobacterium peregrinum
-
analogs with a trivalent nitrogen, competitive inhibition
-
nicotinic acid
nicotinic acid hydrazide
nipecotamide
-
moderate
o-bromobenzamide
-
moderate
p-nitrophenol
-
inhibits amidase and esterase activity
Pyrazinamide
pyrazinecarbonitrile
-
irreversible inactivator
pyrazinoic acid
-
competitive inhibition
pyridine-3-carbonitrile
-
reversible, competitive inhibitor
Sodium diethyl dithiocarbamate
-
-
sulfhydryl reagents
-
-
Thionicotinamide
-
moderate
Triton X-100
-
0.1%, 24% inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
dithiothreitol
Serum albumin
-
0.02%, 9% activation
-
additional information
-
stress suppresses the inhibitory effect of nicotinamide through the induction of PNC1 expression
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.36
5'-methylnicotinamide
-
-
0.55 - 0.68
5-methyl nicotinamide
0.061
5-Methylnicotinamide
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
0.025
Benzamide
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
0.34 - 1.43
ethyl nicotinate
-
0.44 - 1.03
methyl nicotinate
-
0.0002 - 50
nicotinamide
0.016 - 1.2
Pyrazinamide
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.16 - 15.9
5-methyl nicotinamide
1.75
5-Methylnicotinamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
0.0088
Benzamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
0.05 - 0.19
ethyl nicotinate
-
0.17 - 1.02
methyl nicotinate
-
0.0005 - 76.9
nicotinamide
0.1 - 135.7
Pyrazinamide
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0086
1-methylnicotinamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
12283
3.4 - 29.04
5-methyl nicotinamide
206933
0.061 - 29
5-Methylnicotinamide
7130
0.025 - 0.35
Benzamide
1408
0.05 - 0.41
ethyl nicotinate
206935
0.31 - 1.17
methyl nicotinate
206934
0.0096 - 427
nicotinamide
267
0.157 - 331
Pyrazinamide
3666
0.05
Thionicotinamide
Saccharomyces cerevisiae
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
8199
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.37 - 5
2-chloropyridine-3-carbaldehyde
0.316
3-acetylpyridine
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
0.085 - 2
3-Cyanopyridine
0.00029
3-pyridine carboxaldehyde
-
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
0.00044 - 0.153
4-methoxylnicotinaldehyde
0.0044
5-bromonicotinaldehyde
recombinant wild-type enzyme, pH 7.3, 37°C
-
0.00059 - 0.004
5-bromopyridine-3-carbaldehyde
0.000039 - 0.0038
5-methoxypyridine-3-carbaldehyde
0.000023
5-methylnicotinaldehyde
-
in 100 mM phosphate buffer, pH 7.3 at 25°C
0.000023 - 0.00065
5-methylpyridine-3-carbaldehyde
0.004 - 0.018
6-fluoropyridine-3-carbaldehyde
0.0206
benzaldehyde
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
0.000011 - 0.0035
nicotinaldehyde
0.12 - 2
nicotinic acid
6.7
pyrazinoic acid
-
wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C
0.1
pyridine-3-carbonitrile
-
Ki about 0.1 mM, wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.06
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate ethyl nicotinate
0.13
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate ethyl nicotinate
0.22
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate ethyl nicotinate
0.31
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate methyl nicotinate
0.34
purified recombinant mutant C133A, pH 7.3, 37°C, substrate ethyl nicotinate
0.36
purified recombinant mutant K104A, pH 7.3, 37°C, substrate nicotinamide
0.37
purified recombinant mutant T12Q, pH 7.3, 37°C, substrate nicotinamide
0.45
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate methyl nicotinate
0.46
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate pyrazinamide
0.7
purified recombinant mutant E65H, pH 7.3, 37°C, substrate pyrazinamide
1.16
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate methyl nicotinate
1.4
purified recombinant mutant C133A, pH 7.3, 37°C, substrate methyl nicotinate
1.8
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate pyrazinamide
2.17
Flavobacterium peregrinum
-
-
3.62
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate pyrazinamide
3.96
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate 5-methyl nicotinamide
6.39
purified recombinant mutant C133A, pH 7.3, 37°C, substrate pyrazinamide
7.43
purified recombinant mutant F68W/C133A, pH 7.3, 37°C, substrate pyrazinamide
7.5
purified recombinant mutant F68W, pH 7.3, 37°C, substrate pyrazinamide
8.7
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate nicotinamide
9.01
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate 5-methyl nicotinamide
11.74
purified recombinant mutant C133A, pH 7.3, 37°C, substrate 5-methyl nicotinamide
13.5
purified recombinant mutant E65H, pH 7.3, 37°C, substrate nicotinamide
17.5
purified recombinant mutant C133A, pH 7.3, 37°C, substrate nicotinamide
19.97
purified recombinant mutant F68W, pH 7.3, 37°C, substrate nicotinamide
22.9
purified recombinant mutant F68W/C133A, pH 7.3, 37°C, substrate nicotinamide
23.3 - 25
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate nicotinamide
24.8
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate nicotinamide
28
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate 5-methyl nicotinamide
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
Flavobacterium peregrinum
-
-
6.5 - 8.5
-
amidase activity
6.6
Flavobacterium peregrinum
-
-
6.8 - 7.5
-
-
8 - 10
-
esterase
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
activity range, marked decrease in activity below pH 5.0 and above pH 8.0
5 - 10.5
-
pH 5.0: about 70% of maximal activity, pH 10.5: about 75% of maximal activity
5.5 - 9.5
-
pH 5.5: about 60% of maximal activity, pH 9.5: about 45% of maximal activity
6.5 - 10
-
hydrolysis of nicotinamide of a wide range of pH values
7 - 11
-
pH 7.0: about 35% of maximal activity, pH 11.0: about 30% of maximal activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 55
activity range, below 25°C or above 55°C the enzyme loses most of its activity
55 - 80
-
about 35% of maximal activity at 55°C and at 80°C
60 - 100
-
60°C: about 45% of maximal activity, 100°C: 85% of maximal activity
65 - 98
-
65°C: about 50% of maximal activity, 98°C: about 80% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
C1300 neuroblastoma M1 clonal cell line
Manually annotated by BRENDA team
-
embryonic axis
Manually annotated by BRENDA team
-
human erythrocytes infected with Plasmodium falciparum
Manually annotated by BRENDA team
-
activity during germination, overview
Manually annotated by BRENDA team
-
highest level of expression
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
surface location
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Acinetobacter baumannii (strain AYE)
Acinetobacter baumannii (strain AYE)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20890
-
determined by analytical ultracentrifugation and mass spectrometry
21100
2 * 21100, mass spectrometry
21400
Q97PM2
4 * 21400, calculated from amino acid sequence
22900
-
x * 22900, calculated from amino acid sequence
24000
-
x * 24000, SDS-PAGE
26000
-
2 * 26000, SDS-PAGE
34000
-
1 * 34000, SDS-PAGE
39700
dimeric enzyme, gel filtration
42900
gel filtration
43000
-
gel filtration
47000
-
gel filtratrion and analytical ultracentrifugation
65000
-
2 * 65000 + 2 * 50000, SDS-PAGE
211000
-
gel filtration
230000
-
gel electrophoresis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homotetramer
Q97PM2
4 * 21400, calculated from amino acid sequence
monomer
tetramer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
contains 1.7% w/w mannose and 1.5% w/w N-acetylglucosamine
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure (PDB ID 2WT9) analysis, and modeling of substrate binding and unbinding, overview
sitting-drop-based and sparse-matrix screening , using 0.7 M Na-citrate and 0.1 M HEPES (pH 7.5)
-
in complex with nicotinaldehyde, hanging drop vapor diffusion method, using 1.6 M NaOAc, 10% (w/v) ethylene glycol, and 0.1 M HEPES (pH 7.4)
-
resolution 2.9 Angstrom
-
a trapped nicotinoyl-thioester complexed with wild type enzyme and C136S mutant in complex with nicotinamide, hanging drop vapor diffusion method, using 18-22% (w/v) polyethylene glycol 3350, 0.2-0.3 M NaCl and 0.2 M sodium malonate, pH 6.3, at 18°C
Q97PM2
crystal structure (PDB ID 3O94) analysis, and modeling of substrate binding and unbinding, overview
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
-
37°C, 1 h, stable
246570
6 - 8
-
-
679806
6 - 7.3
purified recombinant wild-type enzyme, very stable at pH 6.0 and pH 7.3, where it maintains 40% and 30% residual activity, respectively, after 20 hours of incubation
735084
7 - 9.5
-
37°C, 10 min, stable
246573
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 20
purified recombinant wild-type enzyme, 20 h, stable
40
-
pH 6.8, in presence of Co2+, 10 min, stable up to
55
-
half-life: 2.5 min
65
-
half-life: 1 min
70
-
10 min, complete loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme is stable in buffer solution containing Co2+ and Ni2+
-
freezing and defreezing inactivates
-
inactivated by dialysis in absence of Mn2+. After overnight dialysis in presence of Mn2+ almost all activity is retained. A dialysis against 0.01 M maleate buffer, pH 6.5, containing 0.005 mM HgCl2 results in complete retention of activity
Flavobacterium peregrinum
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
photooxidation of two His residues by methylene blue leads to loss of deaminase activity
-
246572
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 50% glycerol, stable for months
-
-20°C, 50% loss of activity after 4 weeks
Flavobacterium peregrinum
-
0°C, phosphate buffer, pH 7.0, weak loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
immobilized metal affinity chromatography
-
Ni-NTA agarose column chromatography
-
Ni-NTA column chromatography
-
Ni-NTA column chromatography and Sephacryl S-300 gel filtration
-
Ni-NTA resin column chromatography
Q97PM2
nickel chelate chromatography and molecular sieve
-
nickel-based affinity chromatography
-
recombinant enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography, ultrafiltration, and gel filtration
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes methylnicotinamide from Escherichia coli strain Rosetta 2 (DE3) by ultrafiltration and nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a single enzyme accounts for both pyrazinamidase activity and nicotinamidase activity
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli Rosetta 2(DE3) pLysS cells
Q97PM2
expression in Escherichia coli
gene PNC1, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
gene pncA, DNA and amino acid sequence determination and analysis, recombinant enzyme expression in Escherichia coli strain BL21(DE3)pLysS
isolation of a pnc-1a promoter consisting of exon 1a and the intervening region between exon 1a and the adjacent gene, expression as GFP-fusion protein. Expression of His-tagged PNC proteins in Escherichia coli strain BL21(DE3)
-
phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2 (DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C138A
-
significant decrease in enzyme activity
D49A
-
significant decrease in enzyme activity
D8A
-
significant decrease in enzyme activity
H51A
-
significant decrease in enzyme activity
H57A
-
significant decrease in enzyme activity
H57D
-
the mutant has reduced Mn2+ content and shows a 6fold and 38fold decrease in kcat value for nicotinamide and pyrazinamide, respectively
H71A
-
significant decrease in enzyme activity
K96A
-
significant decrease in enzyme activity
S104A
-
partial loss of enzyme activity
S95A
-
partial loss of enzyme activity
H57D
-
the mutant has reduced Mn2+ content and shows a 6fold and 38fold decrease in kcat value for nicotinamide and pyrazinamide, respectively
-
C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
E65H
site-directed mutagenesis, the mutation highly reduces the activity with pyrazinamide compared to the wild-type enzyme
F68W
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
F68W/C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
K104A
site-directed mutagenesis, the mutant shows 98.55% reduced activity compared to the wild-type enzyme
Q96A
site-directed mutagenesis, the mutant shows similar activity with nicotinamide and increased activity with 5-methyl nicotinamide compared to the wild-type enzyme
Q96K
site-directed mutagenesis, the mutant shows 62.7% reduced activity compared to the wild-type enzyme
T12Q
site-directed mutagenesis, the mutant shows 98.42% reduced activity compared to the wild-type enzyme
C133A
-
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
-
K104A
-
site-directed mutagenesis, the mutant shows 98.55% reduced activity compared to the wild-type enzyme
-
Q96A
-
site-directed mutagenesis, the mutant shows similar activity with nicotinamide and increased activity with 5-methyl nicotinamide compared to the wild-type enzyme
-
T12Q
-
site-directed mutagenesis, the mutant shows 98.42% reduced activity compared to the wild-type enzyme
-
D51A
-
the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
D51N
-
the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
D8A
-
the mutant shows 770fold decreased kcat value compared to the wild type enzyme
D8E
-
the mutant shows 100fold decreased kcat value compared to the wild type enzyme
D8N
-
the mutant shows 1000fold decreased kcat value compared to the wild type enzyme
H53A
-
the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
H94A
-
the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme
K122A
-
the mutant shows 16fold decreased kcat value compared to the wild type enzyme
K122R
-
the mutant shows 770fold decreased kcat value compared to the wild type enzyme
C136A
-
the mutant is unable to catalyze nicotinamide hydrolysis
K103A
-
the mutant has only 0.15% of the catalytic activity of the native enzyme
R97A
-
the mutant is a robust nicotinamidase and is nearly as good as the native enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
determination of nicotinamide
drug development
Show AA Sequence (1123 entries)
Please use the Sequence Search for a specific query.