Protein Variants | Comment | Organism |
---|---|---|
F229L | turnover number for adipyl-7-aminodesacetoxycephalosporanic acid is nealy identical to wild-type value, 1.8fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.4fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 1.45fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme | Pseudomonas sp. |
F375L | 1.5fold increase in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 1.2fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 3.9fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 8.2fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme. Improved activity ratio for adipyl-7-aminodesacetoxycephalosporanic acid to glutaryl-7-aminocephalosporanic acid of the mutant enzyme is a consequence of a decreased catalytic efficiency towards glutaryl-7-aminocephalosporanic acid | Pseudomonas sp. |
M271V/Q291K/T374S | 1.3fold increase in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 1.4fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.2fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 1.3fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme | Pseudomonas sp. |
N266H | 1.2fold increase in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 6.9fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.3fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 2fold decrease in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme. Nearly 10fold improved catalytic efficiency on adipyl-7-aminodesacetoxycephalosporanic acid, resulting from a 50% increase in turnover-number and a 6fold decrease in KM-value without decreasing the catalytic efficiency on glutaryl-7-aminocephalosporanic acid | Pseudomonas sp. |
N266S | 1.4fold decrease in turnover number for adipyl-7-aminodesacetoxycephalosporanic acid, 2.3fold decrease in KM-value for adipyl-7-aminodesacetoxycephalosporanic acid, 1.95fold decrease in turnover number for glutaryl-7-aminocephalosporanic acid, 2.8fold increase in Km-value for glutaryl-7-aminocephalosporanic acid as compared to wild-type enzyme | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.016 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266H | Pseudomonas sp. | |
0.033 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, wild-type enzyme | Pseudomonas sp. | |
0.043 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme M271V/Q291K/T374S | Pseudomonas sp. | |
0.048 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme F229L | Pseudomonas sp. | |
0.094 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266S | Pseudomonas sp. | |
0.14 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266H | Pseudomonas sp. | |
0.27 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme F375L | Pseudomonas sp. | |
0.42 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266S | Pseudomonas sp. | |
0.52 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme F229L | Pseudomonas sp. | |
0.7 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme M271V/Q291K/T374S | Pseudomonas sp. | |
0.82 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme F375L | Pseudomonas sp. | |
0.96 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, wild-type enzyme | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | P07662 | SY-77 | - |
Pseudomonas sp. SY-77 | P07662 | SY-77 | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
adipyl-7-aminodesacetoxycephalosporanic acid + H2O | - |
Pseudomonas sp. | deacetoxycephalosporanic acid + adipate | - |
? | |
adipyl-7-aminodesacetoxycephalosporanic acid + H2O | - |
Pseudomonas sp. SY-77 | deacetoxycephalosporanic acid + adipate | - |
? | |
glutaryl-7-aminocephalosporanic acid + H2O | - |
Pseudomonas sp. | 7-aminocephalosporanate + glutarate | - |
? | |
glutaryl-7-aminocephalosporanic acid + H2O | - |
Pseudomonas sp. SY-77 | 7-aminocephalosporanate + glutarate | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.34 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266S | Pseudomonas sp. | |
0.46 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, wild-type enzyme | Pseudomonas sp. | |
0.47 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme F229L | Pseudomonas sp. | |
0.55 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266H | Pseudomonas sp. | |
0.61 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme M271V/Q291K/T374S | Pseudomonas sp. | |
0.67 | - |
adipyl-7-aminodesacetoxycephalosporanic acid | pH 7.5, 37°C, mutant enzyme F375L | Pseudomonas sp. | |
1.1 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme F375L | Pseudomonas sp. | |
2.2 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266S | Pseudomonas sp. | |
3.1 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme F229L | Pseudomonas sp. | |
3.3 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme N266H | Pseudomonas sp. | |
3.5 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, mutant enzyme M271V/Q291K/T374S | Pseudomonas sp. | |
4.3 | - |
glutaryl-7-aminocephalosporanic acid | pH 7.5, 37°C, wild-type enzyme | Pseudomonas sp. |