Protein Variants | Comment | Organism |
---|---|---|
M1A/M16N/M20L/M42Y/C85A/M92F/C152S | mutant ANLYF carries seven amino acid substitutions that result in a methionine- and cysteine-free mutant enzyme with decreased stability. Mutant ANLYF show very high activity and may be stabilized by backbone cyclization via a cyanocysteine-mediated intramolecular ligation reaction, without loss of its high activity | Escherichia coli |
additional information | construction of mutants based on mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, i.e. ANLYF. Linear ANLYF-Gn is ANLYF with the N- and C-terminal extensions of Ala-(Gly)n-3 and -(Gly)2-Cys-Ala residues, respectively, with n=6-8, and circular ANLYF-Gn is ANLYF with the C and N termini cross-linked by the linker of the -Gly-Gly-Ala-(Gly)n-3 sequence, n=6-8 | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0012 | - |
7,8-dihydrofolate | wild-type, 15°C, pH 7.0 | Escherichia coli | |
0.0062 | - |
7,8-dihydrofolate | mutant linear ANLYF-G6, 15°C, pH 7.0 | Escherichia coli | |
0.0083 | - |
7,8-dihydrofolate | mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, 15°C, pH 7.0 | Escherichia coli | |
0.0086 | - |
7,8-dihydrofolate | mutant linear ANLYF-G8, 15°C, pH 7.0 | Escherichia coli | |
0.0115 | - |
7,8-dihydrofolate | mutant circular ANLYF-G6, 15°C, pH 7.0 | Escherichia coli | |
0.013 | - |
7,8-dihydrofolate | mutant circular ANLYF-G7, 15°C, pH 7.0 | Escherichia coli | |
0.014 | - |
7,8-dihydrofolate | mutant circular ANLYF-G8, 15°C, pH 7.0 | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0ABQ4 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | - |
Escherichia coli | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40.9 | - |
melting temperature, mutant linear ANLYF-G7 | Escherichia coli |
42 | - |
melting temperature, mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S | Escherichia coli |
44.3 | - |
melting temperature, mutant linear ANLYF-G8 | Escherichia coli |
44.5 | - |
melting temperature, mutant linear ANLYF-G6 | Escherichia coli |
49.3 | - |
melting temperature, mutant circular ANLYF-G8 | Escherichia coli |
49.5 | - |
melting temperature, mutant circular ANLYF-G6 | Escherichia coli |
50.2 | - |
melting temperature, wild-type | Escherichia coli |
51.3 | - |
melting temperature, mutant circular ANLYF-G7 | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.3 | - |
7,8-dihydrofolate | wild-type, 15°C, pH 7.0 | Escherichia coli | |
34.8 | - |
7,8-dihydrofolate | mutant circular ANLYF-G8, 15°C, pH 7.0 | Escherichia coli | |
36.7 | - |
7,8-dihydrofolate | mutant linear ANLYF-G6, 15°C, pH 7.0 | Escherichia coli | |
38.7 | - |
7,8-dihydrofolate | mutant circular ANLYF-G7, 15°C, pH 7.0 | Escherichia coli | |
40.5 | - |
7,8-dihydrofolate | mutant M1A/M16N/M20L/M42Y/C85A/M92F/C152S, 15°C, pH 7.0 | Escherichia coli | |
40.8 | - |
7,8-dihydrofolate | mutant linear ANLYF-G8, 15°C, pH 7.0 | Escherichia coli | |
41.7 | - |
7,8-dihydrofolate | mutant linear ANLYF-G7, 15°C, pH 7.0 | Escherichia coli | |
43.7 | - |
7,8-dihydrofolate | mutant circular ANLYF-G6, 15°C, pH 7.0 | Escherichia coli |