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Expanding the Substrate Specificity of Thermoanaerobacter pseudoethanolicus Secondary Alcohol Dehydrogenase by a Dual Site Mutation
Author(s) -
Musa Musa M.,
Bsharat Odey,
Karume Ibrahim,
Vieille Claire,
Takahashi Masateru,
Hamdan Samir M.
Publication year - 2018
Publication title -
european journal of organic chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.825
H-Index - 155
eISSN - 1099-0690
pISSN - 1434-193X
DOI - 10.1002/ejoc.201701351
Subject(s) - chemistry , mutant , steric effects , stereochemistry , substrate (aquarium) , alcohol dehydrogenase , enantioselective synthesis , enzyme , active site , biochemistry , catalysis , biology , ecology , gene
Here, we report the asymmetric reduction of selected phenyl‐ring‐containing ketones by various single‐ and dual‐site mutants of Thermoanaerobacter pseudoethanolicus secondary alcohol dehydrogenase (TeSADH). The further expansion of the size of the substrate binding pocket in the mutant W110A/I86A not only allowed the accommodation of substrates of the single mutants W110A and I86A within the expanded active site but also expanded the substrate range of the enzyme to ketones bearing two sterically demanding groups (bulky–bulky ketones), which are not substrates for the TeSADH single mutants. We also report the regio‐ and enantioselective reduction of diketones with W110A/I86A TeSADH and single TeSADH mutants. The double mutant exhibited dual stereopreference to generate the Prelog products most of the time and the anti‐Prelog products in a few cases.