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Protein & Peptide Letters


ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

Modification of Chimeric (2S, 3S)-butanediol Dehydrogenase Based on Structural Information

Author(s): Tomohito Shimegi, Kaito Mochizuki, Takuji Oyama, Takashi Ohtsuki, Masami Kusunoki and Sadaharu Ui

Volume 22 , Issue 3 , 2015

Page: [226 - 233] Pages: 8

DOI: 10.2174/0929866522666150121120823

Price: $65


A chimeric (2S, 3S)-butanediol dehydrogenase (cLBDH) was engineered to have the strict (S)-configuration specificity of the (2S, 3S)-BDH (BsLBDH) derived from Brevibacterium saccharolyticum as well as the enzymatic stability of the (2R, 3S)-BDH (KpMBDH) from Klebsiella pneumonia by swapping the domains of two native BDHs. However, while cLBDH possesses the stability, it lacks the specificity. In order to assist in the design a BDH having strict substrate specificity, an X-ray structural analysis of a cLBDH crystal was conducted at 1.58 Å. The results obtained show some readily apparent differences around the active sites of cLBDH and BsLBDH. Based on this structural information, a novel (2S, 3S)-BDH having a preferred specificity was developed by introducing a V254L mutation into cLBDH. The influence of this mutation on the stability of cLBDH was not evaluated. Nevertheless, the technique described herein is an effective method for the production of a tailor-made BDH.

Keywords: 2, 3-butanediol, butanediol dehydrogenase, domain chimera, short-chain dehydrogenase/reductase family, stereospecificity, tailor-made enzyme, X-ray structural analysis.

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