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Cloning, Expression, and Characterization of budC Gene Encoding meso-2,3-Butanediol Dehydrogenase from Bacillus licheniformis

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Abstract

The budC gene encoding a meso-2,3-butanediol dehydrogenase (BlBDH) from Bacillus licheniformis was cloned and overexpressed in Escherichia coli BL21(DE3). Sequence analysis reveals that this BlBDH belongs to short-chain dehydrogenase/reductase (SDR) superfamily. In the presence of NADH, BlBDH catalyzes the reduction of diacetyl to (3S)-acetoin (97.3 % ee), and further to (2S,3S)-2,3-butanediol (97.3 % ee and 96.5 % de). Similar to other meso-2,3-BDHs, it shows oxidative activity to racemic 2,3-butanediol whereas no activity toward racemic acetoin in the presence of NAD+. For diacetyl reduction and 2,3-butanediol oxidation, the pH optimum of BlBDH is 5.0 and 10.0, respectively. Unusually, it shows relatively high activity over a wide pH range from 5.0 to 8.0 for racemic acetoin reduction. BlBDH shows lower K m and higher catalytic efficiency toward racemic acetoin (K m = 0.47 mM, k cat /K m = 432 s–1·mM–1) when compared with 2,3-butanediol (K m = 7.25 mM, k cat /K m = 81.5 s–1·mM–1), indicating its physiological role in favor of reducing racemic acetoin into 2,3-butanediol. The enzymatic characterization of BlBDH provides evidence for the directed engineering of B. licheniformis for producing enantiopure 2,3-butanediol.

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Acknowledgments

We are grateful to the National Natural Science Foundation of China (21276112 and 21506073), Natural Science Foundation of Jiangsu Province (BK20150003), the Fundamental Research Funds for the Central Universities (JUSRP51409B), the Program of Introducing Talents of Discipline to Universities (111-2-06), and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions for the financial support of this research.

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  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, Jiangsu, China

    Guo-Chao Xu, Ya-Qian Bian, Rui-Zhi Han, Jin-Jun Dong & Ye Ni

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Correspondence to Ye Ni.

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All the authors certify that this manuscript is original and has not been published and will not be submitted elsewhere for publication while being considered by Applied Biochemistry and Biotechnology. Moreover, the study is not split up into several parts to increase the quantity of submissions and submitted to various journals or to one journal over time. No data have been fabricated or manipulated (including images) to support your conclusions. No data, text, or theories by others are presented as if they were our own. The submission has been received explicitly from all coauthors. Furthermore, authors whose names appear on the submission have contributed sufficiently to the scientific work and therefore share collective responsibility and accountability for the results. The authors declare that they have no conflict of interest.

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Xu, GC., Bian, YQ., Han, RZ. et al. Cloning, Expression, and Characterization of budC Gene Encoding meso-2,3-Butanediol Dehydrogenase from Bacillus licheniformis . Appl Biochem Biotechnol 178, 604–617 (2016). https://doi.org/10.1007/s12010-015-1897-3

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