Construction of a Tailor-Made L (2S,3S)-Butanediol Dehydrogenase by Exchanging Domains Between Native Structural Analogs

Tomohito      Shimegi; Yuhsuke      Takusagawa; Takashi      Ohtsuki; Satoko      Noda; Genji      Kurisu; Masami      Kusunoki; Sadaharu      Ui

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: butanediol dehydrogenase, 2, 3-butanediol, domain chimera, short-chain dehydrogenase/reductase family, stereospecificity, tailor-made enzyme, Chiral compounds, liquid crystals, BDH Gene, BDH enzymes, Chimeric L-BDH, Mutagenesis, Structural chimeras, stereoisomers, Domains, Native Structural Analogs, S-configuration, plasmid, E. coli JM109, primer, fermentationbutanediol dehydrogenase, 2, 3-butanediol, domain chimera, short-chain dehydrogenase/reductase family, stereospecificity, tailor-made enzyme, Chiral compounds, liquid crystals, BDH Gene, BDH enzymes, Chimeric L-BDH, Mutagenesis, Structural chimeras, stereoisomers, Domains, Native Structural Analogs, S-configuration, plasmid, E. coli JM109, primer, fermentation