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利用蒸汽辅助浓碱预处理玉米秸秆,工程化谷氨酸棒杆菌高效生产琥珀酸。

Engineering Corynebacterium glutamicum for efficient production of succinic acid from corn stover pretreated by concentrated-alkali under steam-assistant conditions.

机构信息

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.

Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

出版信息

Bioresour Technol. 2023 Jun;378:128991. doi: 10.1016/j.biortech.2023.128991. Epub 2023 Mar 31.

Abstract

Corynebacterium glutamicum was developed for efficient production of succinic acid from corn stover (CS) pretreated by concentrated-alkali under steam-assistant (CASA) conditions. First, C. glutamicum was engineered by 1) blocking the by-products pathways (deletion of ldh, pta-ackA, and cat), 2) enhancing the carbon flux to succinate (overexpression of pyc and ppc), and 3) releasing the end-product inhibition (overexpression of Ncgl0275). The recombinant strain produced 117.8 g/L succinate in fed-batch fermentation. Second, to fully utilize xylose in lignocellulosic hydrolysate, two xylose utilization pathways-the isomerase pathway and the Weimberg pathway-were introduced into the recombinant strain. Third, CS was pretreated by CASA with a higher sugars yield and a lower black liquid. Finally, 64.16 g/L of succinic acid was obtained from 150 g/L CASA-pretreated CS by engineered C. glutamicum. These results showed a succinate high-producing C. glutamicum strain using glucose and xylose simultaneously as well as an effective and environmentally acceptable pretreatment strategy.

摘要

谷氨酸棒杆菌被开发用于从经过蒸汽辅助浓碱预处理的玉米秸秆(CS)中高效生产琥珀酸。首先,通过 1)阻断副产物途径(ldh、pta-ackA 和 cat 的缺失),2)增强碳通量到琥珀酸(pyc 和 ppc 的过表达),3)释放终产物抑制(Ncgl0275 的过表达),对谷氨酸棒杆菌进行工程改造。重组菌在分批补料发酵中生产了 117.8 g/L 的琥珀酸。其次,为了充分利用木质纤维素水解液中的木糖,引入了两条木糖利用途径——异构酶途径和魏因伯格途径——到重组菌中。第三,采用 CASA 对 CS 进行预处理,提高了糖的得率,降低了黑液的含量。最后,通过工程化的谷氨酸棒杆菌从 150 g/L 的 CASA 预处理 CS 中获得了 64.16 g/L 的琥珀酸。这些结果表明,该方法利用葡萄糖和木糖同时生产琥珀酸,同时提供了一种有效且环境友好的预处理策略。

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