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Development of a vitamin B5 hyperproducer in Escherichia coli by multiple metabolic engineering
Metabolic Engineering ( IF 6.8 ) Pub Date : 2024-06-26 , DOI: 10.1016/j.ymben.2024.06.006 Fuqiang Song 1 , Zhijie Qin 2 , Kun Qiu 2 , Zhongshi Huang 2 , Lian Wang 2 , Heng Zhang 2 , Xiaoyu Shan 2 , Hao Meng 3 , Xirong Liu 3 , Jingwen Zhou 1
Metabolic Engineering ( IF 6.8 ) Pub Date : 2024-06-26 , DOI: 10.1016/j.ymben.2024.06.006 Fuqiang Song 1 , Zhijie Qin 2 , Kun Qiu 2 , Zhongshi Huang 2 , Lian Wang 2 , Heng Zhang 2 , Xiaoyu Shan 2 , Hao Meng 3 , Xirong Liu 3 , Jingwen Zhou 1
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Vitamin B [D-pantothenic acid (D-PA)] is an essential water-soluble vitamin that is widely used in the food and feed industries. Currently, the relatively low fermentation efficiency limits the industrial application of D-PA. Here, a plasmid-free D-PA hyperproducer was constructed using systematic metabolic engineering strategies. First, pyruvate was enriched by deleting the non-phosphotransferase system, inhibiting pyruvate competitive branches, and dynamically controlling the TCA cycle. Next, the ()-pantoate pathway was enhanced by screening the rate-limiting enzyme PanBC and regulating the other enzymes of this pathway one by one. Then, to enhance NADPH sustainability, NADPH regeneration was achieved through the novel “PEACES” system by (1) expressing the NAD kinase gene from and the NADP -dependent from and (2) knocking-out the endogenous gene, which interacts with and in the D-PA biosynthesis pathway. Combined with transcriptome analysis, it was found that the membrane proteins OmpC and TolR promoted D-PA efflux by increasing membrane fluidity. Strain PA132 produced a D-PA titer of 83.26 g/L by two-stage fed-batch fermentation, which is the highest D-PA titer reported so far. This work established competitive producers for the industrial production of D-PA and provided an effective strategy for the production of related products.
中文翻译：
通过多重代谢工程在大肠杆菌中开发维生素 B5 高效生产者
维生素 B [D-泛酸 (D-PA)] 是一种必需的水溶性维生素，广泛用于食品和饲料行业。目前，发酵效率较低限制了D-PA的工业化应用。在这里，使用系统代谢工程策略构建了无质粒的 D-PA 超生产者。首先，通过删除非磷酸转移酶系统、抑制丙酮酸竞争性分支、动态控制TCA循环来富集丙酮酸。接下来，通过筛选限速酶PanBC并一一调节该途径的其他酶来增强()-泛解酸途径。然后，为了增强 NADPH 的可持续性，通过新颖的“PEACES”系统实现了 NADPH 再生，方法是（1）表达 NAD 激酶基因和 NADP 依赖性的以及（2）敲除与 NADPH 相互作用的内源基因。 D-PA 生物合成途径。结合转录组分析发现膜蛋白OmpC和TolR通过增加膜流动性促进D-PA外流。菌株PA132通过两阶段补料分批发酵产生的D-PA滴度为83.26 g/L，这是迄今为止报道的最高D-PA滴度。这项工作为D-PA的工业生产建立了有竞争力的生产商，并为相关产品的生产提供了有效的策略。
更新日期：2024-06-26
中文翻译：
通过多重代谢工程在大肠杆菌中开发维生素 B5 高效生产者
维生素 B [D-泛酸 (D-PA)] 是一种必需的水溶性维生素，广泛用于食品和饲料行业。目前，发酵效率较低限制了D-PA的工业化应用。在这里，使用系统代谢工程策略构建了无质粒的 D-PA 超生产者。首先，通过删除非磷酸转移酶系统、抑制丙酮酸竞争性分支、动态控制TCA循环来富集丙酮酸。接下来，通过筛选限速酶PanBC并一一调节该途径的其他酶来增强()-泛解酸途径。然后，为了增强 NADPH 的可持续性，通过新颖的“PEACES”系统实现了 NADPH 再生，方法是（1）表达 NAD 激酶基因和 NADP 依赖性的以及（2）敲除与 NADPH 相互作用的内源基因。 D-PA 生物合成途径。结合转录组分析发现膜蛋白OmpC和TolR通过增加膜流动性促进D-PA外流。菌株PA132通过两阶段补料分批发酵产生的D-PA滴度为83.26 g/L，这是迄今为止报道的最高D-PA滴度。这项工作为D-PA的工业生产建立了有竞争力的生产商，并为相关产品的生产提供了有效的策略。
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