BRENDA - Enzyme Database
show all sequences of 1.14.11.26

Directed evolution of Streptomyces clavuligerus deacetoxycephalosporin C synthase for enhancement of penicillin G expansion

Wei, C.; Yang, Y.; Deng, C.; Liu, W.; Hsu, J.; Lin, Y.; Liaw, S.; Tsai, Y.; Appl. Environ. Microbiol. 71, 8873-8880 (2005)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expressed in Escherichia coli Tuner(DE3) and BL21(DE3) cells
Streptomyces clavuligerus
Engineering
Protein Variants
Commentary
Organism
A106T
80% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
C155Y
90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
C155Y/Y184H/V275I/C281Y
580% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
C281Y
200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
G300V
410% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
G79E
90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
H244Q
140% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
I305L
230% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
I305M
380% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
L277Q
270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
M188I
90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
M188V
150% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
M73T
180% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
N304K
220% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
T91A
110% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
V275I
270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
V275I/I305M
500% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
Y184H
200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.004
-
Penicillin N
mutant enzyme N304K
Streptomyces clavuligerus
0.006
-
Penicillin N
mutant enzyme C281Y; mutant enzyme I305L; mutant enzyme M73T
Streptomyces clavuligerus
0.009
-
Penicillin N
mutant enzyme G79E; mutant enzyme H244Q; mutant enzyme T91A
Streptomyces clavuligerus
0.011
-
Penicillin N
mutant enzyme L277Q; mutant enzyme M188I
Streptomyces clavuligerus
0.012
-
Penicillin N
mutant enzyme I305M; mutant enzyme V275I
Streptomyces clavuligerus
0.013
-
Penicillin N
mutant enzyme A106T; mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.014
-
Penicillin N
wild type enzyme
Streptomyces clavuligerus
0.016
-
Penicillin N
mutant enzyme C155Y
Streptomyces clavuligerus
0.047
-
Penicillin N
mutant enzyme M188V
Streptomyces clavuligerus
0.092
-
Penicillin N
mutant enzyme C155Y/Y184/V275I/C281Y
Streptomyces clavuligerus
0.19
-
penicillin G
mutant enzyme C155Y/Y184/V275I/C281Y
Streptomyces clavuligerus
0.22
-
penicillin G
mutant enzyme N304K
Streptomyces clavuligerus
0.25
-
penicillin G
mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.295
-
Penicillin N
mutant enzyme Y184H
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 M73T
Streptomyces clavuligerus
0.75
-
penicillin G
mutant enzyme G79E; mutant enzyme I305M
Streptomyces clavuligerus
0.76
-
penicillin G
mutant enzyme M188V
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
1.77
-
penicillin G
mutant enzyme A106T
Streptomyces clavuligerus
1.96
-
penicillin G
mutant enzyme M188I
Streptomyces clavuligerus
2.58
-
penicillin G
wild type enzyme
Streptomyces clavuligerus
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
Fe2+-dependent
Streptomyces clavuligerus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
penicillin N + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
? + succinate + CO2
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Streptomyces clavuligerus
-
-
-
Purification (Commentary)
Purification (Commentary)
Organism
anion-exchange chromatography and gel filtration
Streptomyces clavuligerus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
deacetoxycephalosporin C + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
deacetylcephalosporin C + succinate + CO2
-
-
-
?
penicillin G + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
? + succinate + CO2
-
-
-
?
penicillin N + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
? + succinate + CO2
-
-
-
?
phenylacetyl-7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
phenylacetyl-7-aminodeacetylcephalosporin + succinate + CO2
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
DAOCS
-
Streptomyces clavuligerus
deacetoxycephalosporin C synthase
-
Streptomyces clavuligerus
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0315
-
penicillin G
mutant enzyme G79E
Streptomyces clavuligerus
0.0453
-
penicillin G
mutant enzyme A106T; wild type enzyme
Streptomyces clavuligerus
0.0456
-
penicillin G
mutant enzyme M188V
Streptomyces clavuligerus
0.0476
-
penicillin G
mutant enzyme C155Y
Streptomyces clavuligerus
0.048
-
Penicillin N
mutant enzyme C155Y/Y184/V275I/C281Y; mutant enzyme Y184H
Streptomyces clavuligerus
0.0502
-
penicillin G
mutant enzyme V275I
Streptomyces clavuligerus
0.0507
-
penicillin G
mutant enzyme M188I
Streptomyces clavuligerus
0.0522
-
penicillin G
mutant enzyme T91A
Streptomyces clavuligerus
0.0564
-
penicillin G
mutant enzyme N304K
Streptomyces clavuligerus
0.0627
-
penicillin G
mutant enzyme M73T
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
0.084
-
Penicillin N
mutant enzyme M188V
Streptomyces clavuligerus
0.1036
-
penicillin G
mutant enzyme L277Q
Streptomyces clavuligerus
0.1398
-
penicillin G
mutant enzyme C155Y/Y184/V275I/C281Y
Streptomyces clavuligerus
0.1452
-
penicillin G
mutant enzyme I305M
Streptomyces clavuligerus
0.1458
-
penicillin G
mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.178
-
Penicillin N
mutant enzyme G79E
Streptomyces clavuligerus
0.235
-
Penicillin N
mutant enzyme C155Y
Streptomyces clavuligerus
0.239
-
Penicillin N
mutant enzyme M73T
Streptomyces clavuligerus
0.247
-
Penicillin N
mutant enzyme T91A
Streptomyces clavuligerus
0.252
-
Penicillin N
mutant enzyme V275I
Streptomyces clavuligerus
0.26
-
Penicillin N
mutant enzyme H244Q
Streptomyces clavuligerus
0.273
-
Penicillin N
mutant enzyme C281Y
Streptomyces clavuligerus
0.277
-
Penicillin N
mutant enzyme A106T; mutant enzyme M188I
Streptomyces clavuligerus
0.284
-
Penicillin N
mutant enzyme I305L
Streptomyces clavuligerus
0.297
-
Penicillin N
mutant enzyme L277Q
Streptomyces clavuligerus
0.307
-
Penicillin N
wild type enzyme
Streptomyces clavuligerus
0.31
-
Penicillin N
mutant enzyme I305M
Streptomyces clavuligerus
0.316
-
Penicillin N
mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.366
-
Penicillin N
mutant enzyme N304K
Streptomyces clavuligerus
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli Tuner(DE3) and BL21(DE3) cells
Streptomyces clavuligerus
Engineering (protein specific)
Protein Variants
Commentary
Organism
A106T
80% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
C155Y
90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
C155Y/Y184H/V275I/C281Y
580% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
C281Y
200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
G300V
410% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
G79E
90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
H244Q
140% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
I305L
230% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
I305M
380% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
L277Q
270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
M188I
90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
M188V
150% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
M73T
180% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
N304K
220% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
T91A
110% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
V275I
270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
V275I/I305M
500% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
Y184H
200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Streptomyces clavuligerus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.004
-
Penicillin N
mutant enzyme N304K
Streptomyces clavuligerus
0.006
-
Penicillin N
mutant enzyme C281Y; mutant enzyme I305L; mutant enzyme M73T
Streptomyces clavuligerus
0.009
-
Penicillin N
mutant enzyme G79E; mutant enzyme H244Q; mutant enzyme T91A
Streptomyces clavuligerus
0.011
-
Penicillin N
mutant enzyme L277Q; mutant enzyme M188I
Streptomyces clavuligerus
0.012
-
Penicillin N
mutant enzyme I305M; mutant enzyme V275I
Streptomyces clavuligerus
0.013
-
Penicillin N
mutant enzyme A106T; mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.014
-
Penicillin N
wild type enzyme
Streptomyces clavuligerus
0.016
-
Penicillin N
mutant enzyme C155Y
Streptomyces clavuligerus
0.047
-
Penicillin N
mutant enzyme M188V
Streptomyces clavuligerus
0.092
-
Penicillin N
mutant enzyme C155Y/Y184/V275I/C281Y
Streptomyces clavuligerus
0.19
-
penicillin G
mutant enzyme C155Y/Y184/V275I/C281Y
Streptomyces clavuligerus
0.22
-
penicillin G
mutant enzyme N304K
Streptomyces clavuligerus
0.25
-
penicillin G
mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.295
-
Penicillin N
mutant enzyme Y184H
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 M73T
Streptomyces clavuligerus
0.75
-
penicillin G
mutant enzyme G79E; mutant enzyme I305M
Streptomyces clavuligerus
0.76
-
penicillin G
mutant enzyme M188V
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
1.77
-
penicillin G
mutant enzyme A106T
Streptomyces clavuligerus
1.96
-
penicillin G
mutant enzyme M188I
Streptomyces clavuligerus
2.58
-
penicillin G
wild type enzyme
Streptomyces clavuligerus
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
Fe2+-dependent
Streptomyces clavuligerus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
penicillin N + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
? + succinate + CO2
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
anion-exchange chromatography and gel filtration
Streptomyces clavuligerus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
deacetoxycephalosporin C + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
deacetylcephalosporin C + succinate + CO2
-
-
-
?
penicillin G + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
? + succinate + CO2
-
-
-
?
penicillin N + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
? + succinate + CO2
-
-
-
?
phenylacetyl-7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2
-
671443
Streptomyces clavuligerus
phenylacetyl-7-aminodeacetylcephalosporin + succinate + CO2
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0315
-
penicillin G
mutant enzyme G79E
Streptomyces clavuligerus
0.0453
-
penicillin G
mutant enzyme A106T; wild type enzyme
Streptomyces clavuligerus
0.0456
-
penicillin G
mutant enzyme M188V
Streptomyces clavuligerus
0.0476
-
penicillin G
mutant enzyme C155Y
Streptomyces clavuligerus
0.048
-
Penicillin N
mutant enzyme C155Y/Y184/V275I/C281Y; mutant enzyme Y184H
Streptomyces clavuligerus
0.0502
-
penicillin G
mutant enzyme V275I
Streptomyces clavuligerus
0.0507
-
penicillin G
mutant enzyme M188I
Streptomyces clavuligerus
0.0522
-
penicillin G
mutant enzyme T91A
Streptomyces clavuligerus
0.0564
-
penicillin G
mutant enzyme N304K
Streptomyces clavuligerus
0.0627
-
penicillin G
mutant enzyme M73T
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
0.084
-
Penicillin N
mutant enzyme M188V
Streptomyces clavuligerus
0.1036
-
penicillin G
mutant enzyme L277Q
Streptomyces clavuligerus
0.1398
-
penicillin G
mutant enzyme C155Y/Y184/V275I/C281Y
Streptomyces clavuligerus
0.1452
-
penicillin G
mutant enzyme I305M
Streptomyces clavuligerus
0.1458
-
penicillin G
mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.178
-
Penicillin N
mutant enzyme G79E
Streptomyces clavuligerus
0.235
-
Penicillin N
mutant enzyme C155Y
Streptomyces clavuligerus
0.239
-
Penicillin N
mutant enzyme M73T
Streptomyces clavuligerus
0.247
-
Penicillin N
mutant enzyme T91A
Streptomyces clavuligerus
0.252
-
Penicillin N
mutant enzyme V275I
Streptomyces clavuligerus
0.26
-
Penicillin N
mutant enzyme H244Q
Streptomyces clavuligerus
0.273
-
Penicillin N
mutant enzyme C281Y
Streptomyces clavuligerus
0.277
-
Penicillin N
mutant enzyme A106T; mutant enzyme M188I
Streptomyces clavuligerus
0.284
-
Penicillin N
mutant enzyme I305L
Streptomyces clavuligerus
0.297
-
Penicillin N
mutant enzyme L277Q
Streptomyces clavuligerus
0.307
-
Penicillin N
wild type enzyme
Streptomyces clavuligerus
0.31
-
Penicillin N
mutant enzyme I305M
Streptomyces clavuligerus
0.316
-
Penicillin N
mutant enzyme V275I/I305M
Streptomyces clavuligerus
0.366
-
Penicillin N
mutant enzyme N304K
Streptomyces clavuligerus
Other publictions for EC 1.14.11.26
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
-
1
-
53
-
-
5
-
1
-
2
-
3
-
-
-
-
-
-
-
-
4
-
3
1
-
-
6
1
-
-
-
-
-
-
1
-
1
-
-
53
-
-
-
-
5
-
1
-
2
-
-
-
-
-
-
-
-
4
-
1
-
-
6
1
-
-
-
-
4
4
-
6
6
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
-
-
2
-
-
-
-
-
-
2
-
4
-
8
-
-
-
-
-
-
-
-
4
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
2
-
4
-
-
-
-
-
-
-
-
4
-
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