BRENDA - Enzyme Database
show all sequences of 1.14.11.26

Modified deacetylcephalosporin C synthase for the biotransformation of semisynthetic cephalosporins

Balakrishnan, N.; Ganesan, S.; Rajasekaran, P.; Rajendran, L.; Teddu, S.; Durairaaj, M.; Appl. Environ. Microbiol. 82, 3711-3720 (2016)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
ascorbate
required
Streptomyces clavuligerus
Cloned(Commentary)
Cloned (Commentary)
Organism
geen cefF, recombinant exression of cefF library mutants, site-directed mutants, and wild-type enzyme in Escherichia coli strain BL21(DE3)
Streptomyces clavuligerus
Engineering
Protein Variants
Commentary
Organism
A177V
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
A311V
random mutagenesis
Streptomyces clavuligerus
E16G/T90A/T304A
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme; site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
E209Q
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
E82D
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
F267L
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
I193V
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
L236V
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
M184I
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
M229V
random mutagenesis
Streptomyces clavuligerus
additional information
mutagenic cefF library creation by random mutagenesis and screening of the mutant deacetylcephalosporin C synthase enzyme library, homology modeling of DACS structure and mapping of mutant positions. Process-level biotransformation reaction of cephalosporin G to deacetylcephalosporin G by mutants of deacetylcephalosporin C synthase. Deacetylcephalosporin G can be converted completely into hydroxymethyl-7-amino-cephalosporanic acid (HACA) in about 30 min by a subsequent reaction, thus facilitating scalability toward commercialization. Directed-evolution strategies such as random, semirational, rational, and computational methods are used for systematic engineering of DACS for improved activity with cephalosporin G
Streptomyces clavuligerus
P186L
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
P72L
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
R182S
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
R182W
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
S251F
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
S260G
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T273A
random mutagenesis
Streptomyces clavuligerus
T90A
random mutagenesis
Streptomyces clavuligerus
T90A/A311V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/M184I/I193V/F267L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/R182S
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/S251F
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/S260G
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/F267L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/N313D
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/R91G
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/R91G/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/T96S/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/T96S/G255D/A280S
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/V226I
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V221P
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/F195L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/P7L/A237V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/P7L/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/R250L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/V206I/A210V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/V206I/A210V/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V150A
random mutagenesis
Streptomyces clavuligerus
V171L
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V171M
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V221A
random mutagenesis
Streptomyces clavuligerus
V221H
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V221P
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V221T
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V249I
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
Y38C
random mutagenesis
Streptomyces clavuligerus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.58
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V; pH 7.5, 25°C, recombinant wild-type enzyme
Streptomyces clavuligerus
4.41
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
Streptomyces clavuligerus
4.64
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A
Streptomyces clavuligerus
7.24
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
9.2
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
dependent on, required for catalysis
Streptomyces clavuligerus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
deacetoxycephalosporin C + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
deacetylcephalosporin C + succinate + CO2
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
Streptomyces clavuligerus ATCC 27064
-
deacetylcephalosporin C + succinate + CO2
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Streptomyces clavuligerus
P42220
-
-
Streptomyces clavuligerus ATCC 27064
P42220
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
cephalosporin G + 2-oxoglutarate + O2
the wild-type enzyme shows negligible activity toward cephalosporin G, an unnatural and less expensive substrate analogue, but diverse enzyme mutants show increased to high activity with cephalosporin G, overview
744105
Streptomyces clavuligerus
deacetylcephalosporin G + succinate + CO2
-
-
-
?
cephalosporin G + 2-oxoglutarate + O2
the wild-type enzyme shows negligible activity toward cephalosporin G, an unnatural and less expensive substrate analogue, but diverse enzyme mutants show increased to high activity with cephalosporin G, overview
744105
Streptomyces clavuligerus ATCC 27064
deacetylcephalosporin G + succinate + CO2
-
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
-
744105
Streptomyces clavuligerus
deacetylcephalosporin C + succinate + CO2
-
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
-
744105
Streptomyces clavuligerus ATCC 27064
deacetylcephalosporin C + succinate + CO2
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
cefF
-
Streptomyces clavuligerus
DACS
-
Streptomyces clavuligerus
deacetylcephalosporin C synthase
-
Streptomyces clavuligerus
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Streptomyces clavuligerus
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.055
-
cephalosporin G
pH 7.5, 25°C, recombinant wild-type enzyme
Streptomyces clavuligerus
0.122
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V
Streptomyces clavuligerus
0.326
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A
Streptomyces clavuligerus
1.805
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
1.899
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
Streptomyces clavuligerus
2.302
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Streptomyces clavuligerus
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
ascorbate
required
Streptomyces clavuligerus
Cloned(Commentary) (protein specific)
Commentary
Organism
geen cefF, recombinant exression of cefF library mutants, site-directed mutants, and wild-type enzyme in Escherichia coli strain BL21(DE3)
Streptomyces clavuligerus
Engineering (protein specific)
Protein Variants
Commentary
Organism
A177V
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
A311V
random mutagenesis
Streptomyces clavuligerus
E16G/T90A/T304A
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme; site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
E209Q
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
E82D
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
F267L
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
I193V
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
L236V
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
M184I
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
M229V
random mutagenesis
Streptomyces clavuligerus
additional information
mutagenic cefF library creation by random mutagenesis and screening of the mutant deacetylcephalosporin C synthase enzyme library, homology modeling of DACS structure and mapping of mutant positions. Process-level biotransformation reaction of cephalosporin G to deacetylcephalosporin G by mutants of deacetylcephalosporin C synthase. Deacetylcephalosporin G can be converted completely into hydroxymethyl-7-amino-cephalosporanic acid (HACA) in about 30 min by a subsequent reaction, thus facilitating scalability toward commercialization. Directed-evolution strategies such as random, semirational, rational, and computational methods are used for systematic engineering of DACS for improved activity with cephalosporin G
Streptomyces clavuligerus
P186L
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
P72L
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
R182S
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
R182W
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
S251F
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
S260G
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T273A
random mutagenesis
Streptomyces clavuligerus
T90A
random mutagenesis
Streptomyces clavuligerus
T90A/A311V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/M184I/I193V/F267L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/R182S
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/S251F
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/S260G
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/F267L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/N313D
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/R91G
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/R91G/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/T96S/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/T96S/G255D/A280S
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/V226I
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/A40V/M229I/T273A/V221P
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/F195L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/P7L/A237V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/P7L/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/R250L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/V206I/A210V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
T90A/P72L/A311V/V206I/A210V/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V150A
random mutagenesis
Streptomyces clavuligerus
V171L
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V171M
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V221A
random mutagenesis
Streptomyces clavuligerus
V221H
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V221P
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V221T
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
V249I
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
Streptomyces clavuligerus
Y38C
random mutagenesis
Streptomyces clavuligerus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.58
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V; pH 7.5, 25°C, recombinant wild-type enzyme
Streptomyces clavuligerus
4.41
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
Streptomyces clavuligerus
4.64
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A
Streptomyces clavuligerus
7.24
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
9.2
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
dependent on, required for catalysis
Streptomyces clavuligerus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
deacetoxycephalosporin C + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
deacetylcephalosporin C + succinate + CO2
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
Streptomyces clavuligerus ATCC 27064
-
deacetylcephalosporin C + succinate + CO2
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
cephalosporin G + 2-oxoglutarate + O2
the wild-type enzyme shows negligible activity toward cephalosporin G, an unnatural and less expensive substrate analogue, but diverse enzyme mutants show increased to high activity with cephalosporin G, overview
744105
Streptomyces clavuligerus
deacetylcephalosporin G + succinate + CO2
-
-
-
?
cephalosporin G + 2-oxoglutarate + O2
the wild-type enzyme shows negligible activity toward cephalosporin G, an unnatural and less expensive substrate analogue, but diverse enzyme mutants show increased to high activity with cephalosporin G, overview
744105
Streptomyces clavuligerus ATCC 27064
deacetylcephalosporin G + succinate + CO2
-
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
-
744105
Streptomyces clavuligerus
deacetylcephalosporin C + succinate + CO2
-
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
-
744105
Streptomyces clavuligerus ATCC 27064
deacetylcephalosporin C + succinate + CO2
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Streptomyces clavuligerus
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.055
-
cephalosporin G
pH 7.5, 25°C, recombinant wild-type enzyme
Streptomyces clavuligerus
0.122
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V
Streptomyces clavuligerus
0.326
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A
Streptomyces clavuligerus
1.805
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
1.899
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
Streptomyces clavuligerus
2.302
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Streptomyces clavuligerus
General Information
General Information
Commentary
Organism
evolution
the enzyme belongs to the family of 2-oxoglutarate-dependent oxygenases
Streptomyces clavuligerus
metabolism
deacetylcephalosporin C synthase (DACS) transforms an inert methyl group of deacetoxycephalosporin C (DAOC) into an active hydroxyl group of deacetylcephalosporin C (DAC) during the biosynthesis of cephalosporins
Streptomyces clavuligerus
additional information
homology modeling of DACS structure
Streptomyces clavuligerus
physiological function
the enzyme is involved in the biosynthesis of cephalosporins, it carries out a critical transformation of an inert cephem methyl group of deacetoxycephalosporin C (DAOC) into an active hydroxyl group of deacetylcephalosporin C (DAC) in the presence of iron, oxygen, ascorbic acid, and 2-oxoglutarate. It is a step which is chemically difficult to accomplish and which offers a second substitution site for derivatization in cephalosporins
Streptomyces clavuligerus
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme belongs to the family of 2-oxoglutarate-dependent oxygenases
Streptomyces clavuligerus
metabolism
deacetylcephalosporin C synthase (DACS) transforms an inert methyl group of deacetoxycephalosporin C (DAOC) into an active hydroxyl group of deacetylcephalosporin C (DAC) during the biosynthesis of cephalosporins
Streptomyces clavuligerus
additional information
homology modeling of DACS structure
Streptomyces clavuligerus
physiological function
the enzyme is involved in the biosynthesis of cephalosporins, it carries out a critical transformation of an inert cephem methyl group of deacetoxycephalosporin C (DAOC) into an active hydroxyl group of deacetylcephalosporin C (DAC) in the presence of iron, oxygen, ascorbic acid, and 2-oxoglutarate. It is a step which is chemically difficult to accomplish and which offers a second substitution site for derivatization in cephalosporins
Streptomyces clavuligerus
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.021
-
cephalosporin G
pH 7.5, 25°C, recombinant wild-type enzyme
Streptomyces clavuligerus
0.047
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V
Streptomyces clavuligerus
0.07
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A
Streptomyces clavuligerus
0.249
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
0.25
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
0.43
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
Streptomyces clavuligerus
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.021
-
cephalosporin G
pH 7.5, 25°C, recombinant wild-type enzyme
Streptomyces clavuligerus
0.047
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V
Streptomyces clavuligerus
0.07
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A
Streptomyces clavuligerus
0.249
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
0.25
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
Streptomyces clavuligerus
0.43
-
cephalosporin G
pH 7.5, 25°C, recombinant mutant T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
Streptomyces clavuligerus
Other publictions for EC
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
744105
Balakrishnan
Modified deacetylcephalospori ...
Streptomyces clavuligerus, Streptomyces clavuligerus ATCC 27064
Appl. Environ. Microbiol.
82
3711-3720
2016
1
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1
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4
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725639
An
Expression of cefF significant ...
Acremonium chrysogenum, Acremonium chrysogenum CGMCC3.3795, Streptomyces clavuligerus, Streptomyces clavuligerus CGMCC4.1611
J. Ind. Microbiol. Biotechnol.
39
269-274
2012
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3
3
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699366
Goo
Directed evolution and rationa ...
Streptomyces clavuligerus
J. Ind. Microbiol. Biotechnol.
36
619-633
2009
-
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-
-
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-
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700004
Liras
Enzymology of beta-lactam comp ...
Actinomyces sp.
Methods Enzymol.
458
401-429
2009
-
1
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-
-
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3
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671324
Stok
Development of enzyme-linked i ...
Streptomyces clavuligerus
Anal. Chim. Acta
577
153-162
2006
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5
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661796
Wu
C-terminus mutations of Acremo ...
Acremonium chrysogenum
FEMS Microbiol. Lett.
246
103-110
2005
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-
-
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2
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1
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671443
Wei
Directed evolution of Streptom ...
Streptomyces clavuligerus
Appl. Environ. Microbiol.
71
8873-8880
2005
-
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1
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18
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28
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1
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1
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1
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1
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4
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2
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33
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1
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18
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28
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1
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4
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33
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-
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-
659322
Lloyd
Controlling the substrate sele ...
Acremonium chrysogenum
J. Biol. Chem.
279
15420-15426
2004
-
-
-
-
5
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-
-
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1
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4
1
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5
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4
1
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-
440375
Ghag
Refolding and purification of ...
Acremonium chrysogenum
Biotechnol. Appl. Biochem.
24
109-119
1996
-
1
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1
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-
-
-
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-
660772
Coque
Characterization of the cefF g ...
Amycolatopsis lactamdurans
Appl. Microbiol. Biotechnol.
44
605-609
1996
-
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1
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1
1
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662088
Baker
Deacetoxycephalosporin C hydro ...
Streptomyces clavuligerus
J. Biol. Chem.
266
5087-5093
1991
2
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7
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2
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7
1
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440380
Dotzlaf
Purification and properties of ...
Streptomyces clavuligerus
J. Biol. Chem.
264
10219-10227
1989
2
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1
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12
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1
1
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1
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1
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1
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1
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2
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1
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12
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1
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1
1
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1
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1
2
-
-
-
-
-
-
660843
Samson
-
Cloning and expression of the ...
Acremonium chrysogenum
Bio/Technology
5
1207-1214
1987
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660936
Baldwin
Purification and initial chara ...
Acremonium chrysogenum
Biochem. J.
245
831-841
1987
4
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4
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1
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661974
Dotzlaf
Copurification and characteriz ...
Acremonium chrysogenum
J. Bacteriol.
169
1611-1618
1987
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2
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1
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661955
Jensen
Deacetoxycephalosporin C synth ...
Streptomyces clavuligerus
J. Antibiot.
38
263-265
1985
-
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