Highly Efficient Production of N-Acetyl-glucosamine in Escherichia coli by Appropriate Catabolic Division of Labor in the Utilization of Mixed Glycerol/Glucose Carbon Sources,Journal of Agricultural and Food Chemistry

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Highly Efficient Production of N-Acetyl-glucosamine in Escherichia coli by Appropriate Catabolic Division of Labor in the Utilization of Mixed Glycerol/Glucose Carbon Sources
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2021-05-18 , DOI: 10.1021/acs.jafc.1c01513
Qian Ma _{1,

2,

3} , Quanwei Sun ₂ , Miao Tan ₂ , Li Xia ₂ , Ying Zhang ₂ , Mengya Yang ₂ , Mingyang Zhuo ₂ , Kexin Zhao ₂ , Yanjun Li _{1,

2,

3} , Qingyang Xu _{1,

2,

3} , Ning Chen _{1,

2,

3} , Xixian Xie _{1,

2,

3}

Affiliation

Currently, microbial production is becoming a competitive method for N-acetyl-glucosamine production. As the biosynthesis of N-acetyl-glucosamine originating from fructose-6-P directly competes with central carbon metabolism for precursor supply, the consumption of glucose for cell growth and cellular metabolism severely limits the yield of N-acetyl-glucosamine. In this study, appropriate catabolic division of labor in the utilization of mixed carbon sources was achieved by deleting the pfkA gene and enhancing the utilization of glycerol by introducing the glpK mutant. Glycerol thus mainly contributed to cell growth and cellular metabolism, and more glucose was saved for efficient N-acetyl-glucosamine synthesis. By optimizing the ratio of glycerol to glucose, the balancing of cell growth/cellular metabolism and N-acetyl-glucosamine synthesis was achieved. The resulting strain GLALD-7 produced 179.7 g/L N-acetyl-glucosamine using mixed glycerol/glucose (1:8, m/m) carbon sources in a 5 L bioreactor, with a yield of 0.458 g/g total carbon sources (0.529 g/g glucose) and a productivity of 2.57 g/L/h. Coherent high titer/yield/productivity was obtained, with the highest values ever reported, suggesting that an appropriate catabolic division of labor using mixed glycerol/glucose carbon sources is a useful strategy for facilitating the microbial production of chemicals originating from glucose or metabolites upstream of glycolysis.

中文翻译：

利用混合甘油/葡萄糖碳源通过适当的分解代谢分工在大肠杆菌中高效生产N-乙酰氨基葡萄糖

目前，微生物生产正成为N-乙酰氨基葡糖生产的一种竞争方法。由于源自果糖 6-P的N-乙酰氨基葡萄糖的生物合成直接与中央碳代谢竞争前体供应，因此细胞生长和细胞代谢消耗的葡萄糖严重限制了N-乙酰氨基葡萄糖的产量。本研究通过删除pfkA基因并通过引入glpK突变体提高甘油的利用率来实现混合碳源利用中适当的分解代谢分工。因此甘油主要有助于细胞生长和细胞代谢，更多的葡萄糖被节省用于有效的N-乙酰氨基葡萄糖合成。通过优化甘油与葡萄糖的比例，实现了细胞生长/细胞代谢和N-乙酰-氨基葡萄糖合成的平衡。所得菌株 GLALD-7在 5 L 生物反应器中使用混合甘油/葡萄糖 (1:8, m/m) 碳源生产 179.7 g/L N-乙酰氨基葡萄糖，总碳源产量为 0.458 g/g（ 0.529 g/g 葡萄糖）和 2.57 g/L/h 的生产率。获得了一致的高滴度/产量/生产力，具有有史以来报告的最高值，表明使用混合甘油/葡萄糖碳源进行适当的分解代谢分工是促进微生物生产源自葡萄糖或代谢物上游的化学物质的有用策略糖酵解。

更新日期：2021-06-02

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