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Constructing a Synthetic Metabolic Pathway in Escherichia Coli to Produce the Enantiomerically Pure (R, R)-2,3-Butanediol

机译:在大肠杆菌中构建合成代谢途径以生产对映体纯的(R,R)-2,3-丁二醇

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摘要

Enantiomerically pure (R, R)-2,3-butanediol has unique applications due to its special chiral group and spatial configuration. Currently, its chemical production route has many limitations. In addition, no native microorganisms can accumulate (R, R)-2,3-butanediol with an enantio-purity over 99%. Herein, we constructed a synthetic metabolic pathway for enantiomerically pure (R, R)-2,3-butanediol biosynthesis in Escherichia coli. The fermentation results suggested that introduction of the synthetic metabolic pathway redistributed the carbon fluxes to the neutral (R, R)-2,3-butanediol, and thus protected the strain against the acetic acid inhibition. Additionally, it showed that the traditionally used isopropyl beta-D-thiogalactoside (IPTG) induction displayed negative effect on (R, R)-2,3-butanediol biosynthesis in the recombinant E. coli, which was probably due to the protein burden. With no IPTG addition, the (R, R)-2,3-butanediol concentration reached 115g/L by fed-batch culturing of the recombinant E. coli, with an enantio-purity over 99%, which is suitable for the pilot-scale production. Biotechnol. Bioeng. 2015;112: 1056-1059. (c) 2014 Wiley Periodicals, Inc.
机译:对映体纯的(R,R)-2,3-丁二醇由于其特殊的手性基团和空间构型而具有独特的应用。当前,其化学生产路线具有许多局限性。另外,没有天然微生物可以累积对映体纯度超过99%的(R,R)-2,3-丁二醇。本文中,我们构建了对映体纯的(R,R)-2,3-丁二醇在大肠杆菌中生物合成的合成代谢途径。发酵结果表明,合成代谢途径的引入将碳通量重新分配至中性(R,R)-2,3-丁二醇,从而保护了菌株免受乙酸的抑制。此外,它表明,传统上使用的异丙基β-D-硫代半乳糖苷(IPTG)诱导对重组大肠杆菌中(R,R)-2,3-丁二醇的生物合成显示出负面影响,这可能是由于蛋白质负担所致。在不添加IPTG的情况下,通过分批分批培养重组大肠杆菌,(R,R)-2,3-丁二醇浓度达到115g / L,对映体纯度超过99%,适用于中试规模生产。生物技术。生恩2015; 112:1056-1059。 (c)2014年威利期刊有限公司

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