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Literature summary for 3.5.1.93 extracted from

  • Otten, L.G.; Sio, C.F.; Reis, C.R.; Koch, G.; Cool, R.H.; Quax, W.J.
    A highly active adipyl-cephalosporin acylase obtained via rational randomization (2007), FEBS J., 274, 5600-5610.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain DH10B Pseudomonas sp.

Protein Variants

Protein Variants Comment Organism
F375L decreased activity compared to the wild type enzyme Pseudomonas sp.
F375M decreased activity compared to the wild type enzyme Pseudomonas sp.
N266H decreased activity compared to the wild type enzyme Pseudomonas sp.
N266M decreased activity compared to the wild type enzyme Pseudomonas sp.
N266Q decreased activity compared to the wild type enzyme Pseudomonas sp.
N266Q/F375L decreased activity compared to the wild type enzyme Pseudomonas sp.
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.
Y178H decreased activity compared to the wild type enzyme Pseudomonas sp.
Y178H/N266H decreased activity compared to the wild type enzyme Pseudomonas sp.
Y178H/N266M decreased activity compared to the wild type enzyme Pseudomonas sp.
Y178H/N266Q/F375L decreased activity compared to the wild type enzyme Pseudomonas sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.04
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266H Pseudomonas sp.
0.042
-
glutaryl-7-aminocephalosporanic acid wild type enzyme Pseudomonas sp.
0.08
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H Pseudomonas sp.
0.08
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H/N266H Pseudomonas sp.
0.08
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H/N266Q/F375L Pseudomonas sp.
0.12
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266Q Pseudomonas sp.
0.14
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266H Pseudomonas sp.
0.17
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266M Pseudomonas sp.
0.17
-
glutaryl-7-aminocephalosporanic acid mutant enzyme F375M Pseudomonas sp.
0.17
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266Q/F375L Pseudomonas sp.
0.17
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266Q/F375L Pseudomonas sp.
0.17
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H/N266M Pseudomonas sp.
0.19
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H/N266H Pseudomonas sp.
0.26
-
glutaryl-7-aminocephalosporanic acid mutant enzyme F375L Pseudomonas sp.
0.28
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H/N266Q/F375L Pseudomonas sp.
0.3
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H/N266M Pseudomonas sp.
0.33
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266M Pseudomonas sp.
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H Pseudomonas sp.
0.62
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266Q Pseudomonas sp.
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme F375M Pseudomonas sp.
0.7
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme F375L Pseudomonas sp.
0.8
-
adipyl-7-aminodesacetoxycephalosporanic acid wild type enzyme Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp.
-
-
-
Pseudomonas sp. SY-77
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Pseudomonas sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
-
Pseudomonas sp. ?
-
?
adipyl-7-aminodesacetoxycephalosporanic acid + H2O
-
Pseudomonas sp. SY-77 ?
-
?
cephalosporin C + H2O
-
Pseudomonas sp. cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
cephalosporin C + H2O
-
Pseudomonas sp. SY-77 cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
glutaryl-7-aminocephalosporanic acid + H2O
-
Pseudomonas sp. 7-aminocephalosporanate + glutarate
-
?
glutaryl-7-aminocephalosporanic acid + H2O
-
Pseudomonas sp. SY-77 7-aminocephalosporanate + glutarate
-
?

Synonyms

Synonyms Comment Organism
adipyl-cephalosporin acylase
-
Pseudomonas sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.38
-
adipyl-7-aminodesacetoxycephalosporanic acid wild type enzyme Pseudomonas sp.
0.46
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266Q Pseudomonas sp.
0.47
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266H Pseudomonas sp.
0.48
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266Q/F375L Pseudomonas sp.
0.65
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme F375M Pseudomonas sp.
0.66
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H Pseudomonas sp.
0.66
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H/N266Q/F375L Pseudomonas sp.
0.67
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme F375L Pseudomonas sp.
0.75
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266Q/F375L Pseudomonas sp.
0.89
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H/N266M Pseudomonas sp.
0.95
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H/N266H Pseudomonas sp.
1.1
-
glutaryl-7-aminocephalosporanic acid mutant enzyme F375L Pseudomonas sp.
1.2
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme Y178H/N266M Pseudomonas sp.
1.8
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266M Pseudomonas sp.
1.9
-
adipyl-7-aminodesacetoxycephalosporanic acid mutant enzyme N266M Pseudomonas sp.
2.02
-
glutaryl-7-aminocephalosporanic acid mutant enzyme F375M Pseudomonas sp.
2.2
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H/N266H Pseudomonas sp.
2.4
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266Q Pseudomonas sp.
2.5
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H Pseudomonas sp.
2.5
-
glutaryl-7-aminocephalosporanic acid mutant enzyme Y178H/N266Q/F375L Pseudomonas sp.
3.1
-
glutaryl-7-aminocephalosporanic acid mutant enzyme N266H Pseudomonas sp.
4.1
-
glutaryl-7-aminocephalosporanic acid wild type enzyme Pseudomonas sp.