Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | requires the Fe2+ as a cofactor | Streptomyces clavuligerus |
Application | Comment | Organism |
---|---|---|
medicine | antibiotic cephalosporin production | Streptomyces clavuligerus |
Cloned (Comment) | Organism |
---|---|
cloning and heterologous expression in Escherichia coli, Streptomyces lividans, Penicillium chrysogenum, Pseudomonas putida and Pichia pastoris, using of commonly used glutathione S-transferase to express DAOCS as a fusion protein | Streptomyces clavuligerus |
expressed in Escherichia coli, Streptomyces lividans, Penicillium chrysogenum, Pseudomonas putida, and Pichia pastoris | Streptomyces clavuligerus |
the attempt to improve the activity of DAOCS by the directed evolution approach is probably the construction of hybrid DAOCS of Streptomyces clavuligerus and Nocardia lactamdurans using in-vivo homologous recombination | Streptomyces clavuligerus |
Crystallization (Comment) | Organism |
---|---|
- |
Streptomyces clavuligerus |
as apoprotein with N-terminal His tag, 2.3 A resolution | Streptomyces clavuligerus |
as apoprotein, 1.3 A resolution | Streptomyces clavuligerus |
crystallization of mutant delta R306, with Fe2+ and 2-oxoglutarate, 2.1 A resolution | Streptomyces clavuligerus |
crystallization of mutant delta R307A, with Fe2+, succinate and CO2, 1.96 A resolution | Streptomyces clavuligerus |
crystallization with 5-hydroxy-4-ketovaleric acid, 1.53 A resolution | Streptomyces clavuligerus |
crystallization with Fe(II) and succinate , 1.5 A resolution | Streptomyces clavuligerus |
crystallization with Fe(II) and deacetoxycephalosporin C, 1.7 A resolution | Streptomyces clavuligerus |
crystallization with Fe(II) and penicillin G, 1.6 A resolution | Streptomyces clavuligerus |
crystallization with Fe(II), 2-oxoglutarate and ampicillin, 1.5 A resolution | Streptomyces clavuligerus |
crystallization with Fe(II), 2-oxoglutarate and penicillin G, 1.7 A resolution | Streptomyces clavuligerus |
crystallization with Fe2+ and 2-oxoglutarate, 1.5 A resolution, the crystal structure of scDAOCS complexed with 2-oxoglutarate reveals that the 5-carboxylate of 2-oxoglutarate is stabilized by electrostatic interaction with the side chain of R258 | Streptomyces clavuligerus |
crystallization with Fe2+, 1.5 A resolution | Streptomyces clavuligerus |
R258Q mutant, crystallization with Fe(II) and alpha-keto-beta-methylbutanoate, 1.5 and 1.6 A resolution | Streptomyces clavuligerus |
with N-terminal His tag and Fe2+, 2.51 A resolution | Streptomyces clavuligerus |
with N-terminal His tag, crystallization with ampicillin and Fe2+, 2.7 A resolution | Streptomyces clavuligerus |
with N-terminal His tag, crystallization with deacetoxycephalosporin C and Fe2+, 3.0 A resolution | Streptomyces clavuligerus |
Protein Variants | Comment | Organism |
---|---|---|
C155Y | the mutant has 1.5fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
C155Y/Y184H/V275I/C281Y | mutant with enhanced activity and without substrate inhibition | Streptomyces clavuligerus |
C281Y | the mutant has 6.1fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
C281Y/I305M | kcat/Km values of double-mutant scDAOCSs are higher than that of the wild-type enzyme for ampicillin conversion, improvement in the enzymes catalytic effciency (68fold increment) | Streptomyces clavuligerus |
C281Y/N304R | kcat/Km values of double-mutant scDAOCSs are higher than that of the wild-type enzyme for ampicillin conversion, improvement in the enzymes catalytic effciency (101fold increment) | Streptomyces clavuligerus |
G79E | the mutant has 2.3fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
H244Q | the mutant has 2.8fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
I305L | the mutant has 6.4fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
I305M | the mutant has 10.8fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
L277Q | the mutant has 5.7fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
M188V | the mutant has 3.3fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
M73T | the mutant has 4.7fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
N304A | mutation is able to increase the activity of the enzyme | Streptomyces clavuligerus |
N304K | the mutant has 14.2fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
N304R | improvement of the substrate-binding affinity of the enzyme for ampicillin conversion | Streptomyces clavuligerus |
R160L | mutation abolishes the activity of scDAOCS indicating the importance of these residue in binding the penicillin substrate | Streptomyces clavuligerus |
R258L | mutant have the same catalytic activity as the R258Q mutant when 2-oxo-4-methylpentanoate is used as a co-substrate | Streptomyces clavuligerus |
R258Q | mutation in scDAOCS almost abolishes the oxidation of both penicillin N and penicillin G, aliphatic 2-oxoacids (2-oxo-4-methylpentanoate and 2-oxo-3-methylbutanoate), which can not replace the function of 2-oxoglutarate for the wild-type enzyme, are able to rescue the catalytic activity of the R258Q mutant to the level observed for the wild-type enzyme | Streptomyces clavuligerus |
R266L | mutation abolishes the activity of scDAOCS indicating the importance of these residue in binding the penicillin substrate | Streptomyces clavuligerus |
R306I | replacement of R306 with amino acids structurally similar to leucine is able to improve the enzyme activity via hydrophobic interaction with the surrounding residues | Streptomyces clavuligerus |
R306L | mutation improves the conversion activity of scDAOCS | Streptomyces clavuligerus |
R306M | replacement of R306 with amino acids structurally similar to leucine is able to improve the enzyme activity via hydrophobic interaction with the surrounding residues | Streptomyces clavuligerus |
R306P | loss of enzyme activity | Streptomyces clavuligerus |
R74L | mutation abolishes the activity of scDAOCS indicating the importance of these residue in binding the penicillin substrate | Streptomyces clavuligerus |
T91A | the mutant has 2.1fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
V275I | the mutant has 1.7fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
V275I/I305M | the mutant has 32.4fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
Y184H | the mutant has 4.7fold increment in kcat/Km value when compared with the wild type enzyme | Streptomyces clavuligerus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.014 | - |
Penicillin N | wild type enzyme | Streptomyces clavuligerus | |
0.014 | - |
penicillin G | recombinant clone FF8 derived from gene shuffling | Streptomyces clavuligerus | |
0.19 | - |
penicillin G | mutant enzyme C155Y/Y184H/V275I/C281Y | Streptomyces clavuligerus | |
0.22 | - |
penicillin G | mutant enzyme N304K | Streptomyces clavuligerus | |
0.25 | - |
penicillin G | mutant enzyme V275I/I305M | Streptomyces clavuligerus | |
0.66 | - |
penicillin G | mutant enzyme I305L | Streptomyces clavuligerus | |
0.68 | - |
penicillin G | mutant enzyme C281Y | Streptomyces clavuligerus | |
0.74 | - |
penicillin G | mutant enzyme M737T | Streptomyces clavuligerus | |
0.75 | - |
penicillin G | mutant enzyme G79E | Streptomyces clavuligerus | |
0.75 | - |
penicillin G | mutant enzyme I305M | Streptomyces clavuligerus | |
0.76 | - |
penicillin G | mutant enzyme M188V | Streptomyces clavuligerus | |
0.89 | - |
penicillin G | wild type enzyme | Streptomyces clavuligerus | |
0.95 | - |
penicillin G | mutant enzyme Y184H | Streptomyces clavuligerus | |
1.02 | - |
penicillin G | mutant enzyme L277Q | Streptomyces clavuligerus | |
1.36 | - |
penicillin G | mutant enzyme T91A | Streptomyces clavuligerus | |
1.39 | - |
penicillin G | mutant enzyme H244Q | Streptomyces clavuligerus | |
1.68 | - |
penicillin G | mutant enzyme V275I | Streptomyces clavuligerus | |
1.76 | - |
penicillin G | mutant enzyme C155Y | Streptomyces clavuligerus | |
2.58 | - |
penicillin G | wild type enzyme | Streptomyces clavuligerus | |
3.28 | - |
acetyl-6-aminopenicillanic acid | wild type enzyme | Streptomyces clavuligerus | |
4.86 | - |
ampicillin | wild type enzyme | Streptomyces clavuligerus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | acts as cofactor, the iron-binding centre in this family of enzymes essentially provides versatility and flexibility in catalysing a variety of oxidative reactions that include desaturation, ring expansion, and hydroxylation | Streptomyces clavuligerus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2S,5R,6R)-3,3-dimethyl-7-oxo-6-[(thiophen-2-ylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
(6R,7R)-3-methyl-8-oxo-7-[(thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + succinate + CO2 + H2O | - |
? | |
acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-acetylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-adipylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus NRRL 3585 | - |
7-adipylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
amoxicillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? | |
amoxicillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus NRRL 3585 | - |
? | - |
? | |
ampicillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? | |
ampicillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus NRRL 3585 | - |
? | - |
? | |
butyryl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-butyrylaminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
carbenicillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? | |
D-carboxymethylcysteinyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? | |
decanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-decanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
heptanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-heptanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
hexanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-hexanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
metampicillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? | |
additional information | Streptomyces clavuligerus | enzyme is able to convert a wide range of penicillin substrates differing in their side chains, it is a member of 2-oxoglutarate-dependent dioxygenase protein family, the mechanism involves initial activation of the iron centre by 2-oxoglutarate binding followed by oxidative decarboxylation of 2-oxoglutarate to generate oxidizing ferryl species with succinate and carbon dioxide as by-products, succinate remains bound and stabilizes the reactive iron species until the substrate enters the active site, binding of penicillin to the reactive iron species via the sulfur group subsequently expels succinate and leads to oxidative ring expansion of the substrate | ? | - |
? | |
additional information | Streptomyces clavuligerus NRRL 3585 | enzyme is able to convert a wide range of penicillin substrates differing in their side chains, it is a member of 2-oxoglutarate-dependent dioxygenase protein family, the mechanism involves initial activation of the iron centre by 2-oxoglutarate binding followed by oxidative decarboxylation of 2-oxoglutarate to generate oxidizing ferryl species with succinate and carbon dioxide as by-products, succinate remains bound and stabilizes the reactive iron species until the substrate enters the active site, binding of penicillin to the reactive iron species via the sulfur group subsequently expels succinate and leads to oxidative ring expansion of the substrate | ? | - |
? | |
N-nonanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
N7-nonanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-octanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
N7-octanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-valeryl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
N7-pentanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
penicillin F + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-((3E)-hex-3-enoyl)aminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin G + 2-oxoglutarate + O2 | Streptomyces clavuligerus | low catalytic effciency towards penicillin G | 7-phenylacetylaminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin mX + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-[(3-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin N + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
penicillin N + 2-oxoglutarate + O2 | Streptomyces clavuligerus NRRL 3585 | - |
deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
penicillin V + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? | |
penicillin X + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
7-[(4-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
phenethicillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? | |
ticarcillin + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces clavuligerus | P18548 | - |
- |
Streptomyces clavuligerus NRRL 3585 | P18548 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Streptomyces clavuligerus |
affinity chromatography | Streptomyces clavuligerus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
quaternary mutant (C155Y/Y184H/V275I/C281Y) with enhanced activity and without substrate inhibition | Streptomyces clavuligerus |
additional information | - |
the activity of scDAOCS very much depends on the stability of the enzyme maintained by equilibrium in a pool of monomeric and trimeric forms of scDAOCS which can be disrupted by introduction of a relatively small His-tag at the N-terminus of the enzyme | Streptomyces clavuligerus |
additional information | - |
to achieve optimum conditions for the conversion of penicillin to a cephalosporin, a reaction mixture containing 50 mM Tris-HCl (pH 7.4), 10 mM KCl, 10 mM MgSO4, 0.6 mM ascorbic acid, 0.8 mM ATP, 0.04 mM FeSO4, 0.6 mM 2-oxoglutarate, and 0.28 mM penicillin N is used for detection of the ring-expansion activity of DAOCS from Acremonium chrysogenum, DAOCS is unable to convert penicillin G and other penicillin N analogs under this optimum conditions described for penicillin N | Streptomyces clavuligerus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2S,5R,6R)-3,3-dimethyl-7-oxo-6-[(thiophen-2-ylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | (6R,7R)-3-methyl-8-oxo-7-[(thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + succinate + CO2 + H2O | - |
? | |
acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-acetylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-adipylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus NRRL 3585 | 7-adipylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
amoxicillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? | |
amoxicillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus NRRL 3585 | ? | - |
? | |
ampicillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? | |
ampicillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus NRRL 3585 | ? | - |
? | |
butyryl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-butyrylaminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
carbenicillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? | |
D-carboxymethylcysteinyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? | |
decanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-decanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
heptanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-heptanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
hexanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-hexanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
metampicillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? | |
additional information | enzyme is able to convert a wide range of penicillin substrates differing in their side chains, it is a member of 2-oxoglutarate-dependent dioxygenase protein family, the mechanism involves initial activation of the iron centre by 2-oxoglutarate binding followed by oxidative decarboxylation of 2-oxoglutarate to generate oxidizing ferryl species with succinate and carbon dioxide as by-products, succinate remains bound and stabilizes the reactive iron species until the substrate enters the active site, binding of penicillin to the reactive iron species via the sulfur group subsequently expels succinate and leads to oxidative ring expansion of the substrate | Streptomyces clavuligerus | ? | - |
? | |
additional information | enzyme is able to convert a wide range of penicillin substrates differing in their side chains, it is a member of 2-oxoglutarate-dependent dioxygenase protein family, the mechanism involves initial activation of the iron centre by 2-oxoglutarate binding followed by oxidative decarboxylation of 2-oxoglutarate to generate oxidizing ferryl species with succinate and carbon dioxide as by-products, succinate remains bound and stabilizes the reactive iron species until the substrate enters the active site, binding of penicillin to the reactive iron species via the sulfur group subsequently expels succinate and leads to oxidative ring expansion of the substrate | Streptomyces clavuligerus NRRL 3585 | ? | - |
? | |
N-((thiophen-2-yl)acetyl)-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-[(5R,6R)-3,3-dimethyl-2,7-dioxo-4-thia-1-azabicyclo[3.2.0]hept-6-yl]-2-(thiophen-2-yl)acetamide + succinate + CO2 + H2O | - |
? | |
N-acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-acetyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-adipyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-butyryl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-butyryldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-decanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-decanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-heptanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-heptanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-hexanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-hexanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-nonanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N7-nonanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-nonanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-nonanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-octanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N7-octanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-octanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-octanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-valeryl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N7-pentanoyldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
N-valeryl-6-aminopenicillanic acid + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | N-valeryldeacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
penicillin F + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-((3E)-hex-3-enoyl)aminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin G + 2-oxoglutarate + O2 | low catalytic effciency towards penicillin G | Streptomyces clavuligerus | 7-phenylacetylaminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin mX + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-[(3-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus NRRL 3585 | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
penicillin V + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? | |
penicillin X + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | 7-[(4-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O | - |
? | |
phenethicillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? | |
ticarcillin + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | upon addition of Fe(II) and 2-oxoglutarate, scDAOCS dissociates into monomers, determined by gelfiltration and dynamic laser light scattering studies which reveal the existence of an equilibrium between the monomeric and trimeric scDAOCS in the absence of Fe2+ | Streptomyces clavuligerus |
trimer | native state, the C-terminus of one molecule (residues 308-311) penetrates the active site of its neighbouring molecule in a cyclical fashion | Streptomyces clavuligerus |
Synonyms | Comment | Organism |
---|---|---|
DAOCS | - |
Streptomyces clavuligerus |
deacetoxycephalosporin C synthase | - |
Streptomyces clavuligerus |
scDAOCS | - |
Streptomyces clavuligerus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0297 | - |
penicillin G | recombinant FF2 derived from gene shuffling | Streptomyces clavuligerus | |
0.0315 | - |
penicillin G | mutant enzyme G79E | Streptomyces clavuligerus | |
0.0453 | - |
penicillin G | wild type enzyme | Streptomyces clavuligerus | |
0.0456 | - |
penicillin G | mutant enzyme M188V | Streptomyces clavuligerus | |
0.0476 | - |
penicillin G | mutant enzyme C155Y | Streptomyces clavuligerus | |
0.0502 | - |
penicillin G | mutant enzyme V275I | Streptomyces clavuligerus | |
0.0522 | - |
penicillin G | mutant enzyme T91A | Streptomyces clavuligerus | |
0.0564 | - |
penicillin G | mutant enzyme N304K | Streptomyces clavuligerus | |
0.06 | - |
acetyl-6-aminopenicillanic acid | wild type enzyme | Streptomyces clavuligerus | |
0.0627 | - |
penicillin G | mutant enzyme M737T | Streptomyces clavuligerus | |
0.0698 | - |
penicillin G | mutant enzyme H244Q | Streptomyces clavuligerus | |
0.0744 | - |
penicillin G | mutant enzyme C281Y | Streptomyces clavuligerus | |
0.0759 | - |
penicillin G | mutant enzyme I305L | Streptomyces clavuligerus | |
0.0798 | - |
penicillin G | mutant enzyme Y184H | Streptomyces clavuligerus | |
0.1036 | - |
penicillin G | mutant enzyme L277Q | Streptomyces clavuligerus | |
0.12 | - |
ampicillin | wild type enzyme | Streptomyces clavuligerus | |
0.1398 | - |
penicillin G | mutant enzyme C155Y/Y184H/V275I/C281Y | Streptomyces clavuligerus | |
0.1452 | - |
penicillin G | mutant enzyme I305M | Streptomyces clavuligerus | |
0.1458 | - |
penicillin G | mutant enzyme V275I/I305M | Streptomyces clavuligerus | |
0.307 | - |
Penicillin N | wild type enzyme | Streptomyces clavuligerus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
penicillin G | wild type enzyme | Streptomyces clavuligerus | |
0.027 | - |
penicillin G | mutant enzyme C155Y | Streptomyces clavuligerus | |
0.03 | - |
penicillin G | mutant enzyme V275I | Streptomyces clavuligerus | |
0.038 | - |
penicillin G | mutant enzyme T91A | Streptomyces clavuligerus | |
0.042 | - |
penicillin G | mutant enzyme G79E | Streptomyces clavuligerus | |
0.05 | - |
penicillin G | mutant enzyme H244Q | Streptomyces clavuligerus | |
0.06 | - |
penicillin G | mutant enzyme M188V | Streptomyces clavuligerus | |
0.084 | - |
penicillin G | mutant enzyme Y184H | Streptomyces clavuligerus | |
0.085 | - |
penicillin G | mutant enzyme M737T | Streptomyces clavuligerus | |
0.102 | - |
penicillin G | mutant enzyme L277Q | Streptomyces clavuligerus | |
0.109 | - |
penicillin G | mutant enzyme C281Y | Streptomyces clavuligerus | |
0.115 | - |
penicillin G | mutant enzyme I305L | Streptomyces clavuligerus | |
0.194 | - |
penicillin G | mutant enzyme I305M | Streptomyces clavuligerus | |
0.256 | - |
penicillin G | mutant enzyme N304K | Streptomyces clavuligerus | |
0.583 | - |
penicillin G | mutant enzyme V275I/I305M | Streptomyces clavuligerus | |
0.736 | - |
penicillin G | mutant enzyme C155Y/Y184H/V275I/C281Y | Streptomyces clavuligerus |