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Literature summary for 3.5.1.93 extracted from

  • Aramori, I.; Fukagawa, M.; Tsumura, M.; Iwami, M.; Ono, H.; Ishitani, Y.; Koji, H.; Kohsaka, M.; Ueda, Y.; Imanaka, H.
    Comparative characterization of new glutaryl 7-ACA and cephalosporin C acylases (1992), J. Ferment. Bioeng., 73, 185-192.
No PubMed abstract available

Inhibitors

Inhibitors Comment Organism Structure
7-aminocephalosporanate competitive Brevibacillus laterosporus
7-aminocephalosporanate competitive Brevundimonas diminuta
7-aminocephalosporanate competitive Pseudomonas sp.
EDTA 5 mM, 27% inhibition Brevundimonas diminuta
Glutarate competitive Brevibacillus laterosporus
Glutarate competitive Brevundimonas diminuta
Glutarate competitive Pseudomonas sp.
PCMB 92% inhibition at 1 mM, 96% inhibition at 1 mM Brevundimonas diminuta
PCMB 5 mM, 10% inhibition Pseudomonas sp.
PMSF 10% inhibition at 1 mM, 22% inhibition at 5 mM Brevibacillus laterosporus
PMSF 5 mM, 38% inhibition Brevundimonas diminuta

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1
-
glutaryl-7-aminocephalosporanic acid
-
Pseudomonas sp.
2.6
-
glutaryl-7-aminocephalosporanic acid
-
Brevundimonas diminuta
3.2
-
glutaryl-7-aminocephalosporanic acid
-
Brevibacillus laterosporus
4.8
-
cephalosporin C
-
Brevundimonas diminuta

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
1 * 26000 + 1 * 58000, SDS-PAGE Brevundimonas diminuta
28000
-
1 * 28000 + 1 * 61000, SDS-PAGE Pseudomonas sp.
50000
-
gel filtration Brevibacillus laterosporus
54000
-
gel filtration Pseudomonas sp.
54000
-
gel filtration Brevundimonas diminuta
58000
-
1 * 26000 + 1 * 58000, SDS-PAGE Brevundimonas diminuta
61000
-
1 * 28000 + 1 * 61000, SDS-PAGE Pseudomonas sp.
70000
-
1 * 70000, SDS-PAGE Brevibacillus laterosporus

Organism

Organism UniProt Comment Textmining
Brevibacillus laterosporus
-
J1
-
Brevibacillus laterosporus J1
-
J1
-
Brevundimonas diminuta
-
N176
-
Brevundimonas diminuta N176
-
N176
-
Pseudomonas sp.
-
A14, recombinant enzyme
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Pseudomonas sp.
recombinant enzyme Brevibacillus laterosporus
recombinant enzyme Brevundimonas diminuta

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.3
-
-
Brevibacillus laterosporus
7.1
-
-
Pseudomonas sp.
100
-
-
Brevundimonas diminuta

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adipyl-7-aminocephalosporanic acid + H2O activity is 19% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta 7-aminocephalosporanate + adipate
-
?
adipyl-7-aminocephalosporanic acid + H2O activity is 19% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta N176 7-aminocephalosporanate + adipate
-
?
adipyl-7-aminocephalosporanic acid + H2O activity is 0.56% of the activity with glutaryl-7-aminocephalosporanic acid Pseudomonas sp. 7-aminocephalosporanic acid + adipate
-
?
adipyl-7-aminocephalosporanic acid + H2O activity is 68% of the activity with glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus 7-aminocephalosporanic acid + adipate
-
?
adipyl-7-aminocephalosporanic acid + H2O activity is 68% of the activity with glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus J1 7-aminocephalosporanic acid + adipate
-
?
cephalosporin C + H2O weak activity Pseudomonas sp. cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
cephalosporin C + H2O weak activity Brevibacillus laterosporus cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
cephalosporin C + H2O activity is 4% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
cephalosporin C + H2O activity is 4% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta N176 cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
cephalosporin C + H2O weak activity Brevibacillus laterosporus J1 cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
D-glutamyl-7-aminocephalosporanic acid + H2O activity is 0.85% of the activity with glutaryl-7-aminocephalosporanic acid Pseudomonas sp. 7-aminocephalosporanic acid + D-glutamate
-
?
D-glutamyl-7-aminocephalosporanic acid + H2O activity is 0.55% of the activity with glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus 7-aminocephalosporanic acid + glutamate
-
?
D-glutamyl-7-aminocephalosporanic acid + H2O activity is 0.55% of the activity with glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus J1 7-aminocephalosporanic acid + glutamate
-
?
glutaryl-7-aminocephalosporanic acid + H2O rate of synthesis is fold slower than rate of synthesis Brevundimonas diminuta 7-aminocephalosporanate + glutarate
-
r
glutaryl-7-aminocephalosporanic acid + H2O rate of synthesis is slower than rate of hydrolysis Pseudomonas sp. 7-aminocephalosporanate + glutarate
-
r
glutaryl-7-aminocephalosporanic acid + H2O rate of synthesis is slower than rate of hydrolysis Brevibacillus laterosporus 7-aminocephalosporanate + glutarate
-
r
glutaryl-7-aminocephalosporanic acid + H2O rate of synthesis is fold slower than rate of synthesis Brevundimonas diminuta N176 7-aminocephalosporanate + glutarate
-
r
glutaryl-7-aminocephalosporanic acid + H2O rate of synthesis is slower than rate of hydrolysis Brevibacillus laterosporus J1 7-aminocephalosporanate + glutarate
-
r
N-acetyl cephalosporin C + H2O activity is 1% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta acetate + cephalosporin C
-
?
N-acetyl cephalosporin C + H2O activity is 1% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta N176 acetate + cephalosporin C
-
?
succinyl-7-aminocephalosporanic acid + H2O activity is 32% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta 7-aminocephalosporanate + succinate
-
?
succinyl-7-aminocephalosporanic acid + H2O activity is 32% of the activity with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta N176 7-aminocephalosporanate + succinate
-
?
succinyl-7-aminocephalosporanic acid + H2O activity is 6.1% of the activity with glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus 7-aminocephalosporanic acid + succinate
-
?
succinyl-7-aminocephalosporanic acid + H2O activity is 9.9fold higher than with glutaryl-7-aminocephalosporanic acid Pseudomonas sp. 7-aminocephalosporanic acid + succinate
-
?
succinyl-7-aminocephalosporanic acid + H2O activity is 6.1% of the activity with glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus J1 7-aminocephalosporanic acid + succinate
-
?

Subunits

Subunits Comment Organism
? 1 * 70000, SDS-PAGE Brevibacillus laterosporus
? 1 * 26000 + 1 * 58000, SDS-PAGE Brevundimonas diminuta
dimer 1 * 28000 + 1 * 61000, SDS-PAGE Pseudomonas sp.

Synonyms

Synonyms Comment Organism
cephalosporin C acylase
-
Pseudomonas sp.
cephalosporin C acylase
-
Brevibacillus laterosporus
cephalosporin C acylase
-
Brevundimonas diminuta

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Brevibacillus laterosporus
40
-
-
Brevundimonas diminuta
48
-
-
Pseudomonas sp.

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 55 30°C: about 45% of maximal activity, 55°C: about 70% of maximal activity Pseudomonas sp.
30 55 30°C: about 45% of maximal activity, 55°C: about 70% of maximal activity Brevibacillus laterosporus
30 55 30°C: about 45% of maximal activity, 55°C: about 70% of maximal activity Brevundimonas diminuta

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
1 h, less than 10% loss of activity at pH 6.0, about 60% less of activity at pH 5, about 35% loss of activity at pH 7, about 50% loss of activity at pH 8, about 90% loss of activity at pH 9 Pseudomonas sp.
50
-
1 h, less than 10% loss of activity at pH 7.0, about 20% less of activity at pH 5, about 35% loss of activity at pH 7, about 70% loss of activity at pH 8, about 90% loss of activity at pH 9 Brevibacillus laterosporus
50
-
1 h, stable at pH 9.0, about 45% less of activity at pH 6, about 15% loss of activity at pH 7, about 10% loss of activity at pH 8, about 60% loss of activity at pH 10 Brevundimonas diminuta

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
reaction with glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus
9
-
reaction with glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta
9 10 reaction with glutaryl-7-aminocephalosporanic acid Pseudomonas sp.

pH Range

pH Minimum pH Maximum Comment Organism
6 9 pH 6: about 50% of maximal activity, pH 90: about 90% of maximal activity, hydrolysis of glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus
7 9 activity decrease from pH 7 to pH 9, synthesis of glutaryl-7-aminocephalosporanic acid Pseudomonas sp.
7 9 activity decreases from pH 7 to pH 9, synthesis of glutaryl-7-aminocephalosporanic acid Brevibacillus laterosporus
8 10 pH 8: about 50% of maximal activity, pH 9-10: maximal activity, hydrolysis of glutaryl-7-aminocephalosporanic acid Pseudomonas sp.
8
-
activity decrease from pH 7 to pH 9, synthesis of glutaryl-7-aminocephalosporanic acid Brevundimonas diminuta

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.4
-
7-aminocephalosporanate
-
Brevundimonas diminuta
2
-
7-aminocephalosporanate
-
Brevibacillus laterosporus
2.3
-
7-aminocephalosporanate
-
Pseudomonas sp.
2.5
-
Glutarate
-
Brevundimonas diminuta
10
-
Glutarate
-
Pseudomonas sp.
53
-
Glutarate
-
Brevibacillus laterosporus

pI Value

Organism Comment pI Value Maximum pI Value
Brevundimonas diminuta isoelectric focusing
-
4.5
Brevibacillus laterosporus isoelectric focusing
-
4.7
Pseudomonas sp. above, isoelectric focusing
-
6.5