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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
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 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 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. |