BRENDA - Enzyme Database

A highly active adipyl-cephalosporin acylase obtained via rational randomization

Otten, L.G.; Sio, C.F.; Reis, C.R.; Koch, G.; Cool, R.H.; Quax, W.J.; FEBS J. 274, 5600-5610 (2007) View publication on PubMed

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
3.5.1.93
expressed in Escherichia coli strain DH10B
Pseudomonas sp.
Protein Variants
EC Number
Protein Variants
Commentary
Organism
3.5.1.93
F375L
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
F375M
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266H
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266M
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266Q
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266Q/F375L
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178F/F375H
the mutation synergistically improve the catalytic efficiency towards adipyl-7-ADCA 36fold, The activity of this double mutant towards adipyl-7-ADCA is 50% of the activity of the wild-type enzyme towards the preferred substrate glutaryl-7-aminocephalosporanic acid
Pseudomonas sp.
3.5.1.93
Y178H
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178H/N266H
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178H/N266M
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178H/N266Q/F375L
decreased activity compared to the wild type enzyme
Pseudomonas sp.
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.1.93
0.04
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
0.042
-
glutaryl-7-aminocephalosporanic acid
wild type enzyme
Pseudomonas sp.
3.5.1.93
0.08
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
0.08
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
0.08
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.12
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
0.14
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
0.17
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
0.17
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
0.17
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.17
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.17
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
0.19
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
0.26
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
0.28
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.3
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
0.33
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
0.62
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
0.7
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
0.8
-
adipyl-7-aminodesacetoxycephalosporanic acid
wild type enzyme
Pseudomonas sp.
Organism
EC Number
Organism
UniProt
Commentary
Textmining
3.5.1.93
Pseudomonas sp.
-
-
-
3.5.1.93
Pseudomonas sp. SY-77
-
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
3.5.1.93
-
Pseudomonas sp.
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
3.5.1.93
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
-
686677
Pseudomonas sp.
?
-
-
-
?
3.5.1.93
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
-
686677
Pseudomonas sp. SY-77
?
-
-
-
?
3.5.1.93
cephalosporin C + H2O
-
686677
Pseudomonas sp.
cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
-
-
?
3.5.1.93
cephalosporin C + H2O
-
686677
Pseudomonas sp. SY-77
cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
-
-
?
3.5.1.93
glutaryl-7-aminocephalosporanic acid + H2O
-
686677
Pseudomonas sp.
7-aminocephalosporanate + glutarate
-
-
-
?
3.5.1.93
glutaryl-7-aminocephalosporanic acid + H2O
-
686677
Pseudomonas sp. SY-77
7-aminocephalosporanate + glutarate
-
-
-
?
Synonyms
EC Number
Synonyms
Commentary
Organism
3.5.1.93
adipyl-cephalosporin acylase
-
Pseudomonas sp.
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.1.93
0.38
-
adipyl-7-aminodesacetoxycephalosporanic acid
wild type enzyme
Pseudomonas sp.
3.5.1.93
0.46
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
0.48
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
0.66
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
0.66
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.67
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
0.75
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.89
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
0.95
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
1.1
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
1.2
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
1.8
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
1.9
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
2.02
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
2.2
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
2.4
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
2.5
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
2.5
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
3.1
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
4.1
-
glutaryl-7-aminocephalosporanic acid
wild type enzyme
Pseudomonas sp.
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.93
expressed in Escherichia coli strain DH10B
Pseudomonas sp.
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
3.5.1.93
F375L
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
F375M
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266H
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266M
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266Q
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
N266Q/F375L
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178F/F375H
the mutation synergistically improve the catalytic efficiency towards adipyl-7-ADCA 36fold, The activity of this double mutant towards adipyl-7-ADCA is 50% of the activity of the wild-type enzyme towards the preferred substrate glutaryl-7-aminocephalosporanic acid
Pseudomonas sp.
3.5.1.93
Y178H
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178H/N266H
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178H/N266M
decreased activity compared to the wild type enzyme
Pseudomonas sp.
3.5.1.93
Y178H/N266Q/F375L
decreased activity compared to the wild type enzyme
Pseudomonas sp.
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.1.93
0.04
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
0.042
-
glutaryl-7-aminocephalosporanic acid
wild type enzyme
Pseudomonas sp.
3.5.1.93
0.08
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
0.08
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
0.08
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.12
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
0.14
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
0.17
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
0.17
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
0.17
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.17
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.17
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
0.19
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
0.26
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
0.28
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.3
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
0.33
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
0.62
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
0.7
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
0.8
-
adipyl-7-aminodesacetoxycephalosporanic acid
wild type enzyme
Pseudomonas sp.
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.93
-
Pseudomonas sp.
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
3.5.1.93
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
-
686677
Pseudomonas sp.
?
-
-
-
?
3.5.1.93
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
-
686677
Pseudomonas sp. SY-77
?
-
-
-
?
3.5.1.93
cephalosporin C + H2O
-
686677
Pseudomonas sp.
cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
-
-
?
3.5.1.93
cephalosporin C + H2O
-
686677
Pseudomonas sp. SY-77
cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
-
-
?
3.5.1.93
glutaryl-7-aminocephalosporanic acid + H2O
-
686677
Pseudomonas sp.
7-aminocephalosporanate + glutarate
-
-
-
?
3.5.1.93
glutaryl-7-aminocephalosporanic acid + H2O
-
686677
Pseudomonas sp. SY-77
7-aminocephalosporanate + glutarate
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.1.93
0.38
-
adipyl-7-aminodesacetoxycephalosporanic acid
wild type enzyme
Pseudomonas sp.
3.5.1.93
0.46
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
0.48
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
0.66
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
0.66
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.67
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
0.75
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q/F375L
Pseudomonas sp.
3.5.1.93
0.89
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
0.95
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
1.1
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375L
Pseudomonas sp.
3.5.1.93
1.2
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme Y178H/N266M
Pseudomonas sp.
3.5.1.93
1.8
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
1.9
-
adipyl-7-aminodesacetoxycephalosporanic acid
mutant enzyme N266M
Pseudomonas sp.
3.5.1.93
2.02
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme F375M
Pseudomonas sp.
3.5.1.93
2.2
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266H
Pseudomonas sp.
3.5.1.93
2.4
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266Q
Pseudomonas sp.
3.5.1.93
2.5
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H
Pseudomonas sp.
3.5.1.93
2.5
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme Y178H/N266Q/F375L
Pseudomonas sp.
3.5.1.93
3.1
-
glutaryl-7-aminocephalosporanic acid
mutant enzyme N266H
Pseudomonas sp.
3.5.1.93
4.1
-
glutaryl-7-aminocephalosporanic acid
wild type enzyme
Pseudomonas sp.