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Engineering a Microbial Consortium Based Whole-Cell System for Efficient Production of Glutarate From L-Lysine

机译:基于微生物联盟的全细胞系统工程设计可从L-赖氨酸高效生产谷氨酸

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摘要

Glutarate is an important C5 platform chemical produced during the catabolism of L-lysine through 5-aminovalerate (5-AMV) pathway. Here, we first established a whole-cell biocatalysis system for the glutarate production from L-lysine with the engineered Escherichia coli (E. coli) that co-expressed DavAB and GabDT. However, the accumulation of intermediate 5-AMV was identified as one important factor limiting glutarate production. Meanwhile, the negative interaction of co-expressing DavAB and GabDT in a single cell was also confirmed. Here, we solved these problems through engineering a microbial consortium composed of two engineered E. coli strains, BL21-22AB and BL21-YDT, as the whole-cell biocatalysts, each of which contains a part of the glutarate pathway. After the optimization of bioconversion conditions, including temperature, metal ion additives, pH, and cell ratio, 17.2 g/L glutarate was obtained from 20 g/L L-lysine with a yield of 95.1%, which was improved by 19.2% compared with that in a single cell. Little accumulation of 5-AMV was detected. Even at the high substrate concentration, the reduced 5-AMV accumulation and increased glutarate production were achieved. This synthetic consortium produced 43.8 g/L glutarate via a fed-batch strategy, the highest titer reported to date.
机译:谷氨酸盐是L-赖氨酸通过5-氨基戊酸(5-AMV)途径分解代谢过程中产生的重要C5平台化学品。在这里,我们首先建立了一个全细胞生物催化系统,用于从L-赖氨酸与共表达DavAB和GabDT的工程化大肠杆菌(E. coli)生产谷氨酸。然而,中间体5-AMV的积累被认为是限制谷氨酸生产的重要因素。同时,还证实了在单个细胞中共表达DavAB和GabDT的负相互作用。在这里,我们通过工程改造由两个工程大肠杆菌菌株BL21-22AB和BL21-YDT组成的微生物联盟作为全细胞生物催化剂,解决了这些问题,它们每个都包含一部分谷氨酸途径。在优化了温度,金属离子添加剂,pH和细胞比例等生物转化条件后,从20 g / L L-赖氨酸中获得了17.2 g / L谷氨酸,产率为95.1%,与上一年度相比提高了19.2%。在一个单元格中。检测到5-AMV很少积累。即使在高底物浓度下,也能减少5-AMV积累并增加谷氨酸产量。该合成财团通过分批补料策略生产了43.8 g / L的谷氨酸盐,是迄今为止报道的最高滴度。

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