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Literature summary for 3.1.1.6 extracted from

  • Bartsch, S.; Kourist, R.; Bornscheuer, U.T.
    Complete inversion of enantioselectivity towards acetylated tertiary alcohols by a double mutant of a Bacillus subtilis esterase (2008), Angew. Chem. Int. Ed. Engl., 47, 1508-1511.
    View publication on PubMed

Crystallization (Commentary)

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

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

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms Comment Organism
BS2
-
Bacillus subtilis