文献类型：期刊文献

英文题名：Cloning, expression and characterization of glycerol dehydrogenase involved in 2,3-butanediol formation in Serratia marcescens H30

作者：Zhang, Liaoyuan[1];Xu, Quanming[1];Peng, Xiaoqian[1];Xu, Boheng[1];Wu, Yuehao[1];Yang, Yulong[1];Sun, Shujing[1];Hu, Kaihui[1];Shen, Yaling[2]

机构：[1]Fujian Agr & Forestry Univ, Key Lab Biopesticide & Chem Biol, Coll Life Sci, Gutian Edible Fungi Res Inst,Minist Educ, Fuzhou 350002, Fujian, Peoples R China;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, New World Inst Biotechnol, Shanghai 200237, Peoples R China

年份：2014

卷号：41

期号：9

起止页码：1319

外文期刊名：JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY

收录：;EI(收录号：20242616454762);WOS:【SCI-EXPANDED(收录号:WOS:000340559500001)】；

基金：This work was supported by the National Natural Science Foundation of China (No. 31301858), Research Fund for the Doctoral Program of Higher Education of China (No. 20133515120011), the Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Serratia marcescens; 2,3-Butanediol isomers; Glycerol dehydrogenase; Expression; Enzymatic properties

摘要：The meso-2,3-butanediol dehydrogenase (meso-BDH) from S. marcescens H30 is responsible for converting acetoin into 2,3-butanediol during sugar fermentation. Inactivation of the meso-BDH encoded by budC gene does not completely abolish 2,3-butanediol production, which suggests that another similar enzyme involved in 2,3-butanediol formation exists in S. marcescens H30. In the present study, a glycerol dehydrogenase (GDH) encoded by gldA gene from S. marcescens H30 was expressed in Escherichia coli BL21(DE3), purified and characterized for its properties. In vitro conversion indicated that the purified GDH could catalyze the interconversion of (3S)-acetoin/meso-2,3-butanediol and (3R)-acetoin/(2R,3R)-2,3-butanediol. (2S,3S)-2,3-Butanediol was not a substrate for the GDH at all. Kinetic parameters of the GDH enzyme showed lower K (m) value and higher catalytic efficiency for (3S/3R)-acetoin in comparison to those for (2R,3R)-2,3-butanediol and meso-2,3-butanediol, implying its physiological role in favor of 2,3-butanediol formation. Maximum activity for reduction of (3S/3R)-acetoin and oxidations of meso-2,3-butanediol and glycerol was observed at pH 8.0, while it was pH 7.0 for diacetyl reduction. The enzyme exhibited relative high thermotolerance with optimum temperature of 60 A degrees C in the oxidation-reduction reactions. Over 60 % of maximum activity was retained at 70 A degrees C. Additionally, the GDH activity was significantly enhanced for meso-2,3-BD oxidation in the presence of Fe2+ and for (3S/3R)-acetoin reduction in the presence of Mn2+, while several cations inhibited its activity, particularly Fe2+ and Fe3+ for (3S/3R)-acetoin reduction. The properties provided potential application for single configuration production of acetoin and 2,3-butanediol .