Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.1.93 extracted from

  • Saito, Y.; Fujimura, T.; Ishii, Y.; Noguchi, Y.; Miura, T.; Niwa, M.; Shimomura, K.
    Oxidative modification of a cephalosporin C acylase from Pseudomonas strain N176 and site-directed mutagenesis of the gene (1996), Appl. Environ. Microbiol., 62, 2919-2925.
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
M116A at pH 7.5, 76.9% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 95.8% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 108% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate Pseudomonas sp.
M157A at pH 7.5, 70.9% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 58% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 65% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is lower than that of wild-type enzyme Pseudomonas sp.
M164A at pH 7.5, 167% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 104% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 86.6% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is higher than that of wild-type enzyme. The ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.2fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 2.16fold higher than the wild-type ratio Pseudomonas sp.
M164F the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.3fold lower than the wild-type ratio Pseudomonas sp.
M164G the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.3fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 2.23fold higher than the wild-type ratio Pseudomonas sp.
M164L the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 2.1fold lower than the wild-type ratio Pseudomonas sp.
M164N the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.4fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is 1.81fold higher than the wild-type ratio Pseudomonas sp.
M164P the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.3fold lower than the wild-type ratio Pseudomonas sp.
M164Q the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 5.3fold lower than the wild-type ratio. The ratio of turnover number to KM-value for glutaryl-7-aminocephalosporanic acid is nearly identical to the wild-type ratio Pseudomonas sp.
M164S the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.5fold lower than the wild-type ratio Pseudomonas sp.
M164T the ratio of specific activity with cephalosporin C as substrate to that with glutaryl-7-aminocephalosporanic acid as substrate is 1.4fold lower than the wild-type ratio Pseudomonas sp.
M174A at pH 7.5, 96.5% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 109% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 122% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate Pseudomonas sp.
M227A at pH 7.5, 106% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 74.2% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 105% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate. Stability after treatment with H2O2 is lower than that of wild-type enzyme Pseudomonas sp.
M98A at pH 7.5, 126% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 83.1% of the specific activity measured with wild-type enzyme with glutaryl-7-aminocephalosporanic acid as substrate. At pH 8.7, 91.2% of the specific activity measured with wild-type enzyme with cephalosporin C as substrate Pseudomonas sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164A Pseudomonas sp.
2.57
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164G Pseudomonas sp.
3.68
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164N Pseudomonas sp.
5.85
-
glutaryl-7-aminocephalosporanic acid 37°C, wild-type enzyme Pseudomonas sp.
9.32
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164Q Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp.
-
-
-

Substrates and Products (Substrate)

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
pH 8.0, 6 h, 25 mM Tris-HCl, 22% inactivation of wild-type enzyme, 26% inactivation of mutant enzyme M164A, 29% inactivation of mutant enzyme M164G, 20% inactivation of mutant enzyme M164L, 29% inactivation of mutant enzyme M164N, 10% inactivation of mutant enzyme M164Q Pseudomonas sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
8.29
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164Q Pseudomonas sp.
15.1
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164N Pseudomonas sp.
18.1
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164A Pseudomonas sp.
18.6
-
glutaryl-7-aminocephalosporanic acid 37°C, mutant enzyme M164G Pseudomonas sp.
49.8
-
glutaryl-7-aminocephalosporanic acid 37°C, wild-type enzyme Pseudomonas sp.