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Literature summary for 3.5.1.93 extracted from

  • Pollegioni, L.; Lorenzi, S.; Rosini, E.; Marcone, G.L.; Molla, G.; Verga, R.; Cabri, W.; Pilone, M.S.
    Evolution of an acylase active on cephalosporin C (2005), Protein Sci., 14, 3064-3076.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
synthesis bioconversion of cephalosporin C into 7-aminocephalosporanate Pseudomonas sp.

Cloned(Commentary)

Cloned (Comment) Organism
expression of mutant enzymes in Escherichia coli Pseudomonas sp.

Protein Variants

Protein Variants Comment Organism
A215E Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 7.6fold lower than wild-type value Pseudomonas sp.
A215F Vmax/Km for cephalosporin C is 2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 3.9fold lower than wild-type value Pseudomonas sp.
A215L Vmax/Km for cephalosporin C is 1.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 4.2fold lower than wild-type value Pseudomonas sp.
A215V Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.5fold lower than wild-type value Pseudomonas sp.
A215Y Vmax/Km for cephalosporin C is 4.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.6fold lower than wild-type value Pseudomonas sp.
A215Y/H296S Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 25.1fold lower than wild-type value Pseudomonas sp.
A215Y/H296S/H309S Vmax/Km for cephalosporin C is 3.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 13.7fold lower than wild-type value Pseudomonas sp.
A215Y/H309S Vmax/Km for cephalosporin C is 3.8fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 2.9fold higher than wild-type value Pseudomonas sp.
D416Y Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.2fold lower than wild-type value Pseudomonas sp.
D416Y/H417Y Vmax/Km for cephalosporin C is 5.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 151fold lower than wild-type value Pseudomonas sp.
E89A/A215Y Vmax/Km for cephalosporin C is 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 25.2fold lower than wild-type value Pseudomonas sp.
F270M Vmax/Km for cephalosporin C is 1.8fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.7fold lower than wild-type value Pseudomonas sp.
H296F Vmax/Km for cephalosporin C is 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 50.3fold lower than wild-type value Pseudomonas sp.
H296N Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is fold lower than wild-type value Pseudomonas sp.
H296S Vmax/Km for cephalosporin C is 1.7fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 10.1fold lower than wild-type value Pseudomonas sp.
H296S/H309S Vmax/Km for cephalosporin C is 4fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 21.6fold lower than wild-type value Pseudomonas sp.
H296S/H417M Vmax/Km for cephalosporin C is 2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 151fold lower than wild-type value Pseudomonas sp.
H296T Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.2fold lower than wild-type value Pseudomonas sp.
H309S Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 8.9fold lower than wild-type value Pseudomonas sp.
H309Y Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 16.8fold lower than wild-type value Pseudomonas sp.
H417Y Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 4.4fold lower than wild-type value Pseudomonas sp.
I44V/E49stop/D416Y/H417Y Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is more than 1510fold lower than wild-type value Pseudomonas sp.
S22P/T394P/D416Y/H417Y Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is more than 1510fold lower than wild-type value Pseudomonas sp.
Y215Y/F270S Vmax/Km for cephalosporin C is 2.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.2fold lower than wild-type value Pseudomonas sp.
Y271F Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.4fold lower than wild-type value Pseudomonas sp.
Y27V Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 2.1fold lower than wild-type value Pseudomonas sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.7
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296S Pseudomonas sp.
0.9
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215F Pseudomonas sp.
0.9
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y/H309S Pseudomonas sp.
1
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215E Pseudomonas sp.
1
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H309Y Pseudomonas sp.
1.1
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme E89A/A215Y Pseudomonas sp.
1.2
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215V Pseudomonas sp.
1.2
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme Y271F Pseudomonas sp.
1.6
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, wild-type enzyme Pseudomonas sp.
1.7
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215L Pseudomonas sp.
1.7
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y Pseudomonas sp.
1.7
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme D416Y Pseudomonas sp.
1.7
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H296S/H417M Pseudomonas sp.
1.9
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296F Pseudomonas sp.
1.9
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme Y27V Pseudomonas sp.
2
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296T Pseudomonas sp.
2
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme Y215Y/F270S Pseudomonas sp.
2.5
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y/H296S/H309S Pseudomonas sp.
2.7
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme F270M Pseudomonas sp.
2.7
-
cephalosporin C pH 8.0, 37°C, mutant enzyme Y271V Pseudomonas sp.
2.8
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296N Pseudomonas sp.
3.2
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215E Pseudomonas sp.
4
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y/H296S Pseudomonas sp.
4.1
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H296F Pseudomonas sp.
4.3
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H417Y Pseudomonas sp.
4.6
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H309S Pseudomonas sp.
4.8
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H296T Pseudomonas sp.
5.2
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H296N Pseudomonas sp.
5.5
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296S/H417M Pseudomonas sp.
6.3
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H309Y Pseudomonas sp.
6.5
-
cephalosporin C pH 8.0, 37°C, mutant enzyme F270M Pseudomonas sp.
6.7
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H296S Pseudomonas sp.
6.9
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y Pseudomonas sp.
6.9
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296S/H309S Pseudomonas sp.
7.2
-
cephalosporin C pH 8.0, 37°C, mutant enzyme D416Y Pseudomonas sp.
7.7
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215F Pseudomonas sp.
8
-
cephalosporin C pH 8.0, 37°C, mutant enzyme Y271F Pseudomonas sp.
8.3
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215V Pseudomonas sp.
9
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme D416Y/H417Y Pseudomonas sp.
9.5
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215L Pseudomonas sp.
10
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y/H296S Pseudomonas sp.
10.1
-
cephalosporin C pH 8.0, 37°C, mutant enzyme Y215Y/F270S Pseudomonas sp.
11
-
cephalosporin C pH 8.0, 37°C, wild-type enzyme Pseudomonas sp.
12.2
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H296S/H309S Pseudomonas sp.
12.3
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H417Y Pseudomonas sp.
13
-
cephalosporin C pH 8.0, 37°C, mutant enzyme D416Y/H417Y Pseudomonas sp.
17.2
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y/H309S Pseudomonas sp.
17.8
-
cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y/H296S/H309S Pseudomonas sp.
18.9
-
cephalosporin C pH 8.0, 37°C, mutant enzyme E89A/A215Y Pseudomonas sp.
24.4
-
cephalosporin C pH 8.0, 37°C, mutant enzyme H309S Pseudomonas sp.
24.5
-
cephalosporin C pH 8.0, 37°C, mutant enzyme S22P/T394P/D416Y/H417Y Pseudomonas sp.
25
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme I44V/E49stop/D416Y/H417Y Pseudomonas sp.
32
-
cephalosporin C pH 8.0, 37°C, mutant enzyme I44V/E49stop/D416Y/H417Y Pseudomonas sp.
37.2
-
glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme S22P/T394P/D416Y/H417Y Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp.
-
N176
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cephalosporin C + H2O
-
Pseudomonas sp. cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
glutaryl-7-amino cephalosporanic acid + H2O
-
Pseudomonas sp. 7-aminocephalosporanate + glutarate
-
?