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在工程菌中构建具有NADH/NAD再生系统的用于生产2,3-丁二醇/乙偶姻的高效全细胞生物催化剂

Efficient whole-cell biocatalyst for 2,3-butanediol/acetoin production with NADH/NAD regeneration system in engineered .

作者信息

Ben Rao, Tu Truong Ngoc, Ying Fan Ji, An Sun Jian, Shen Yaling

机构信息

State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology, Shanghai 200237, P.R.China.

出版信息

J Microbiol Biotechnol. 2017 May 24. doi: 10.4014/jmb.1612.12057.

Abstract

is reported to possess the potential for industrial 2,3-butanediol or acetoin production by fermentation. But 2,3-butanediol or acetoin are always co-produced and this may make purification process difficult. So biocatalytic technologies may be the appropriate production methods. In this study, we developed an auto-inducing expression system based on pET system and quorum sensing system by using . By using this system, could be engineered as whole-cell biocatalyst to obtain 2,3-butanediol or acetoin. In order to convert diacetyl to 2,3-butanediol, formate dehydrogenase (FDH) and 2,3-butanediol dehydrogenase were co-expressed to construct a NADH regeneration system. This whole-cell biocatalyst could efficiently produced 53.6 g/L 2,3-butanediol with a productivity of 3.35 g/Lh. Next, in order to convert 2,3-butanediol to acetoin, NADH oxidase and 2,3-butanediol dehydrogenase were co-expressed to construct a NAD+ regeneration system. This whole-cell biocatalyst could efficiently produced 59 g/L acetoin with a productivity of 2.95 g/Lh. This work indicated this auto-inducing system was a powerful tool to construct whole-cell biocatalyst in in 2,3-butanediol and acetoin production.

摘要

据报道,其具有通过发酵生产工业2,3 - 丁二醇或乙偶姻的潜力。但是2,3 - 丁二醇或乙偶姻总是共同产生,这可能会使纯化过程变得困难。因此,生物催化技术可能是合适的生产方法。在本研究中,我们通过使用……开发了一种基于pET系统和群体感应系统的自诱导表达系统。通过使用该系统,……可被工程化为全细胞生物催化剂以获得2,3 - 丁二醇或乙偶姻。为了将双乙酰转化为2,3 - 丁二醇,共表达甲酸脱氢酶(FDH)和2,3 - 丁二醇脱氢酶以构建NADH再生系统。这种全细胞生物催化剂能够高效地生产53.6 g/L的2,3 - 丁二醇,生产率为3.35 g/Lh。接下来,为了将2,3 - 丁二醇转化为乙偶姻,共表达NADH氧化酶和2,3 - 丁二醇脱氢酶以构建NAD⁺再生系统。这种全细胞生物催化剂能够高效地生产59 g/L的乙偶姻,生产率为2.95 g/Lh。这项工作表明,这种自诱导系统是在2,3 - 丁二醇和乙偶姻生产中构建全细胞生物催化剂的有力工具。

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