Liu Ran, Zhu Fayin, Lu Lei, Fu Aisi, Lu Jiankai, Deng Zixin, Liu Tiangang
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and Wuhan University School of Pharmaceutical Sciences, Wuhan 430071, China; Wuhan J1 Biotech Co., Ltd., Wuhan 430075, China.
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, and Wuhan University School of Pharmaceutical Sciences, Wuhan 430071, China.
Metab Eng. 2014 Mar;22:10-21. doi: 10.1016/j.ymben.2013.12.004. Epub 2013 Dec 12.
Fatty alcohols are important components of surfactants and cosmetic products. The production of fatty alcohols from sustainable resources using microbial fermentation could reduce dependence on fossil fuels and greenhouse gas emission. However, the industrialization of this process has been hampered by the current low yield and productivity of this synthetic pathway. As a result of metabolic engineering strategies, an Escherichia coli mutant containing Synechococcus elongatus fatty acyl-ACP reductase showed improved yield and productivity. Proteomics analysis and in vitro enzymatic assays showed that endogenous E. coli AdhP is a major contributor to the reduction of fatty aldehydes to fatty alcohols. Both in vitro and in vivo results clearly demonstrated that the activity and expression level of fatty acyl-CoA/ACP reductase is the rate-limiting step in the current protocol. In 2.5-L fed-batch fermentation with glycerol as the only carbon source, the most productive E. coli mutant produced 0.75 g/L fatty alcohols (0.02 g fatty alcohol/g glycerol) with a productivity of up to 0.06 g/L/h. This investigation establishes a promising synthetic pathway for industrial microbial production of fatty alcohols.
脂肪醇是表面活性剂和化妆品的重要成分。利用微生物发酵从可持续资源生产脂肪醇可以减少对化石燃料的依赖并降低温室气体排放。然而,该合成途径目前的低产量和低生产率阻碍了这一过程的工业化。通过代谢工程策略,含有聚球藻脂肪酸酰基-ACP还原酶的大肠杆菌突变体的产量和生产率得到了提高。蛋白质组学分析和体外酶活性测定表明,内源性大肠杆菌AdhP是将脂肪醛还原为脂肪醇的主要贡献者。体外和体内结果均清楚表明,脂肪酸酰基辅酶A/ACP还原酶的活性和表达水平是当前方案中的限速步骤。在以甘油作为唯一碳源的2.5升补料分批发酵中,产量最高的大肠杆菌突变体产生了0.75克/升脂肪醇(0.02克脂肪醇/克甘油),生产率高达0.06克/升/小时。本研究为工业微生物生产脂肪醇建立了一条有前景的合成途径。