Application | Comment | Organism |
---|---|---|
synthesis | mutant enzyme M269Y may be a promising enzyme in the one-step enzymic production of 7-amino-cephalosporanic acid from cephalosporin C | Pseudomonas sp. |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli. Semi-large-scale fermentation of recombinant enzyme | Pseudomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
A271F | 118% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 56.2% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
A271L | 104% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 100% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
A271Y | 101% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 122% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K100Q | 81% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 106% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K114Q | 86% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 101% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K170Q | 130% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 95.6% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K187Q | 113% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 91.1% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K255Q | 107% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 97% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K301Q | 101% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate | Pseudomonas sp. |
K44Q | 102% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 111% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K507Q | 102% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 113.9% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
K629Q | 94.2% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate | Pseudomonas sp. |
K73Q | 46.9% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 47% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
M269F | 98.2% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 165% of wild-type activity with cephalosporin C as substrate. The ratio of turnover number to Km-value with cephalosporin C as substrate is 1.8fold higher than the wild-type ratio | Pseudomonas sp. |
M269L | 92.5% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 107.9% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
M269Y | 91.5% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 155% of wild-type activity with cephalosporin C as substrate. The ratio of turnover number to Km-value with cephalosporin C as substrate is 1.6fold higher than the wild-type ratio | Pseudomonas sp. |
S293C | 1.21% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, no activity with cephalosporin C as substrate | Pseudomonas sp. |
Y270A | 70.6% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 24.3% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
Y270E | no activity with glutaryl-7-aminocephalosporanic acid or cephalosporin C as substrate | Pseudomonas sp. |
Y270F | 50.4% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 46.1% of wild-type activity with cephalosporin C as substrate. The ratio of turnover number to Km-value with cephalosporin C as substrate is 2.6fold lower than the wild-type ratio | Pseudomonas sp. |
Y270L | 28.1% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 32.3% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
Y270S | 61.7% of wild-type activity with glutaryl-7-aminocephalosporanic acid as substrate, 28.3% of wild-type activity with cephalosporin C as substrate | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
21.3 | - |
cephalosporin C | 37°C, mutant enzyme Y269F | Pseudomonas sp. | |
21.6 | - |
cephalosporin C | 37°C, wild-type enzyme | Pseudomonas sp. | |
22.1 | - |
cephalosporin C | 37°C, mutant enzyme Y269Y | Pseudomonas sp. | |
27.8 | - |
cephalosporin C | 37°C, mutant enzyme Y270F | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cephalosporin C + H2O | - |
Pseudomonas sp. | 7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate | - |
? | |
glutaryl-7-aminocephalosporanic acid + H2O | - |
Pseudomonas sp. | 7-aminocephalosporanic acid + glutarate | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.91 | - |
cephalosporin C | 37°C, mutant enzyme Y270F | Pseudomonas sp. | |
5.93 | - |
cephalosporin C | 37°C, wild-type enzyme | Pseudomonas sp. | |
9.4 | - |
cephalosporin C | 37°C, mutant enzyme Y269Y | Pseudomonas sp. | |
10.4 | - |
cephalosporin C | 37°C, mutant enzyme Y269F | Pseudomonas sp. |