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

[ Vol. 18 , Issue. 8 ]

Author(s):

Tomohito Shimegi, Yuhsuke Takusagawa, Takashi Ohtsuki, Satoko Noda, Genji Kurisu, Masami Kusunoki and Sadaharu Ui   Pages 825 - 830 ( 6 )

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

Affiliation:

Department of Biotechnology,Graduate School of Medicine and Engineering, Univ. of Yamanashi, Takeda,Kofu 400-8511, Japan.



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