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在一氧化碳上生长的丙酮丁醇梭菌中的乙醇代谢动力学。

Ethanol Metabolism Dynamics in Clostridium ljungdahlii Grown on Carbon Monoxide.

机构信息

Shandong Provincial Key Laboratory of Synthetic Biology, Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, People's Republic of China.

Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, People's Republic of China.

出版信息

Appl Environ Microbiol. 2020 Jul 2;86(14). doi: 10.1128/AEM.00730-20.

Abstract

Bioethanol production from syngas using acetogenic bacteria has attracted considerable attention in recent years. However, low ethanol yield is the biggest challenge that prevents the commercialization of syngas fermentation into biofuels using microbial catalysts. The present study demonstrated that ethanol metabolism plays an important role in recycling NADH/NAD during autotrophic growth. Deletion of bifunctional aldehyde/alcohol dehydrogenase () genes leads to significant growth deficiencies in gas fermentation. Using specific fermentation technology in which the gas pressure and pH were constantly controlled at 0.1 MPa and 6.0, respectively, we revealed that ethanol was formed during the exponential phase, closely accompanied by biomass production. Then, ethanol was oxidized to acetate via the aldehyde ferredoxin oxidoreductase pathway in A metabolic experiment using C-labeled ethanol and acetate, redox balance analysis, and comparative transcriptomic analysis demonstrated that ethanol production and reuse shared the metabolic pathway but occurred at different growth phases. Ethanol production from carbon monoxide (CO) as a carbon and energy source by and "" is currently being commercialized. During gas fermentation, ethanol synthesis is NADH-dependent. However, ethanol oxidation and its regulatory mechanism remain incompletely understood. Energy metabolism analysis demonstrated that reduced ferredoxin is the sole source of NADH formation by the Rnf-ATPase system, which provides ATP for cell growth during CO fermentation. Therefore, ethanol production is tightly linked to biomass production (ATP production). Clarification of the mechanism of ethanol oxidation and biosynthesis can provide an important reference for generating high-ethanol-yield strains of in the future.

摘要

近年来,利用产乙酸菌从合成气生产生物乙醇引起了人们的极大关注。然而,乙醇产量低是最大的挑战,这阻碍了利用微生物催化剂将合成气发酵转化为生物燃料的商业化进程。本研究表明,乙醇代谢在自养生长过程中回收 NADH/NAD 中起着重要作用。双功能醛/醇脱氢酶()基因的缺失导致气体发酵中的生长缺陷显著。通过使用特定的发酵技术,将气体压力和 pH 值分别恒定控制在 0.1 MPa 和 6.0,我们揭示了乙醇在指数生长期形成,与生物量的产生密切相关。然后,通过醛铁氧还蛋白氧化还原酶途径,乙醇被氧化为乙酸。通过 C 标记的乙醇和乙酸的代谢实验、氧化还原平衡分析和比较转录组分析表明,乙醇的产生和再利用共享代谢途径,但发生在不同的生长阶段。目前正在商业化利用产乙酸菌()和()从一氧化碳(CO)作为碳和能源源生产乙醇。在气体发酵过程中,乙醇的合成依赖于 NADH。然而,乙醇的氧化及其调控机制仍不完全清楚。能量代谢分析表明,还原型铁氧还蛋白是 Rnf-ATPase 系统形成 NADH 的唯一来源,该系统在 CO 发酵过程中为细胞生长提供 ATP。因此,乙醇的生产与生物量的生产(ATP 的产生)紧密相关。阐明乙醇氧化和生物合成的机制可以为未来生成具有高乙醇产量的菌株提供重要参考。

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