Crystallization (Comment) | Organism |
---|---|
molecular dynamic simulations and automated docking with R and S enantiomers of substrate 1,1,1-trifluoro-2-phenyl-but-3-yn-1-yl acetate using structure of wild-type and structural model of double mutant E188W/M193C, S enantiomer fits better in active site of mutant E188W/M193C because subtrates phenyl group points out of it, docking of the preferred R enantiomer by the wild-type with lower free energy than docking of the S enantiomer | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
E188D | very low specific activity, R enantioselectivity of above 100, S enantioselectivity of 26, mutant generated using a focused directed-evolution approach based on saturation mutagenesis according to CASTing method | Bacillus subtilis |
E188W | very low specific activity similar to mutant M193C, S enantioselectivity of 26 | Bacillus subtilis |
E188W/M193C | 14% of wild-type specific activity, wider substrate range than wild-type with respect to tertiary alcohol acetates, inversed enantioselectivity, S enantioselectivity of 64, binding of S enantiomer of substrate 1,1,1-trifluoro-2-phenyl-but-3-yn-1-yl acetate with lower free energy than wild-type, M193C mutation seems to stabilize S orientation, mutant generated using a focused directed-evolution approach based on saturation mutagenesis according to CASTing method | Bacillus subtilis |
M193C | very low specific activity similar to mutant E188W, R enantioselectivity of 16 | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
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Bacillus subtilis | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
46.5 | - |
mutant E188W/M193C, using 4-nitrophenyl acetate as substrate | Bacillus subtilis |
333 | - |
wild-type, using 4-nitrophenyl acetate as substrate | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-phenyl-1-(trifluoromethyl)prop-2-yn-1-yl acetate + H2O | determination of enantiopreference, wild-type R enantioselectivity is 42 | Bacillus subtilis | 1-phenyl-1-(trifluoromethyl)prop-2-yn-1-ol + acetate | spectrophotometric assay coupling released acetate to NADH generation | ? | |
4-nitrophenyl acetate + H2O | determination of specific activity | Bacillus subtilis | 4-nitrophenol + acetate | - |
? | |
additional information | 1-(4-fluorophenyl)-1-(trifluoromethyl)prop-2-yn-1-yl acetate, 1-(4-chlorophenyl)-1-(trifluoromethyl)prop-2-yn-1-yl acetate, 1-(4-methylphenyl)-1-(trifluoromethyl)prop-2-yn-1-yl acetate, 1-(3-fluorophenyl)-1-(trifluoromethyl)prop-2-yn-1-yl acetate, 1-(3,4-difluorophenyl)-1-(trifluoromethyl)prop-2-yn-1-yl acetate are substrates of double mutant E188W/M193C, conversion with inversed enantioselectivity | Bacillus subtilis | ? | - |
? | |
additional information | 1-phenyl-1-(trifluoromethyl)but-2-yn-1-yl acetate and 1-phenyl-1-(trifluoromethyl)prop-2-en-1-yl acetate are no substrates of neither wild-type nor any mutant | Bacillus subtilis | ? | - |
? | |
additional information | size of residue at position 188 (from Asp to Trp) determines enantioselectivity and -preference | Bacillus subtilis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BS2 | - |
Bacillus subtilis |