Construction of a Tailor-Made L (2S,3S)-Butanediol Dehydrogenase by Exchanging Domains Between Native Structural Analogs
Abstract:The development of a stable L-BDH chimera was attempted by exchanging whole domains between two native structural analogs, L-BDH and meso-BDH, because the S-configuration specificity of L-BDH is valuable from the standpoint of its application but its activity is unstable, whereas meso-BDH is stable. The domain chimeras obtained indicated that the leaf-like structures constituting three domains were likely to be mainly associated with chiral recognition, and the fourth domain, the basic domain, is likely to be mainly associated with enzyme stability. A combination of the leaf domains of L-BDH and the basic domain of meso-BDH attained a sufficient level of practical use as an artificial L-BDH chimera, because the resulting enzyme had both stability and S-configuration specificity. However, the levels of stability and specificity were slightly lower than those of the respective enzymes from which they were derived.
Keywords: 2,3-butanediol; BDH Gene; BDH enzymes; Chimeric L-BDH; Chiral compounds; Domains; E. coli JM109; Mutagenesis; Native Structural Analogs; S-configuration; Structural chimeras; butanediol dehydrogenase; domain chimera; fermentation; liquid crystals; plasmid; primer; short-chain dehydrogenase/reductase family; stereoisomers; stereospecificity; tailor-made enzyme
Document Type: Research Article
Publication date: August 1, 2011
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