Cloned (Comment) | Organism |
---|---|
gene hlacz-sh, chemical synthesis of the gene derived from Pyrococcus woesei for direct evolution, DNA and amino acid sequence determination and anaylsis, library construction, expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta-Blue (DE3) | Pyrococcus woesei |
Protein Variants | Comment | Organism |
---|---|---|
T29A/V213I/L217M/N277H/I387V/R491C/N496D | large range mutant screening after direct chemical evolution, the mutant variant YG6762 has nucleotide changes in 13 sites: T30C, A85G, C621A, G637A, T649A, A829V, T912G, C924T, A1159G, G1242C, C1471T, A1486G, and C1497A, the mutant YG6762 shows 4.9-7.5fold increased activity, dependent on the reaction temperature, but decreased thermostability, compared to the wild-type enzyme, overview, revertion of the mutation at each single of the 8 mutation sites results in decreased activity of the altered mutant, especially when reverting residue 277 to Asn, overview | Pyrococcus woesei |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus woesei | O52629 | wild-type gene hlacz-sh; gene hlacz-sh | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-nitrophenyl beta-D-galactopyranoside + H2O | - |
Pyrococcus woesei | 2-nitrophenol + beta-D-galactose | - |
? | |
4-nitrophenyl beta-D-galactopyranoside + H2O | - |
Pyrococcus woesei | 4-nitrophenol + beta-D-galactose | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure analysis and molecular modeling, overview | Pyrococcus woesei |
Synonyms | Comment | Organism |
---|---|---|
YH4502 | - |
Pyrococcus woesei |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
90 | - |
- |
Pyrococcus woesei |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
100 | - |
the mutant enzyme YG6762 shows 40% remaining activity after 1 h, and 30% remaining activity after 2 h, wild-type enzyme YH4502 shows over 75% remaining actiivty after 1 h, and more than 50% remiaing activity after 2 h | Pyrococcus woesei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
mutant YG6762 | Pyrococcus woesei |
5.5 | - |
wild-type enzyme YH4502 | Pyrococcus woesei |