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

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

Volume: 22 Issue: 3

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

Affiliation:

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

Abstract: 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.

Cite this article as:

Shimegi Tomohito, Mochizuki Kaito, Oyama Takuji, Ohtsuki Takashi, Kusunoki Masami and Ui Sadaharu, Modification of Chimeric (2S, 3S)-butanediol Dehydrogenase Based on Structural Information, Protein & Peptide Letters 2015; 22 (3) . https://dx.doi.org/10.2174/0929866522666150121120823