OBJECTIVES To establish a technique for efficient fatty acid production through enhancement of coenzyme A (CoA) biosynthesis and malonyl-CoA supply by introducing exogenous pantothenate kinase (coaA) and acetyl-CoA carboxylase (acc) in Escherichia coli. RESULTS The expression of acc, obtained from Corynebacterium glutamicum, accumulated 2.2-fold more fatty acids in E. coli. The addition of coaA from Pseudomonas putaida or fatty acid synthase (fasA) from C. glutamicum resulted in a 3.1- and 3.6-fold increase in fatty acid synthesis in E. coli cells, which expressed acc and coaA, or acc and fasA, respectively. The transformants, simultaneously possessing all three genes, produced 5.6-fold more fatty acids. The strain possessing acc, coaA, and fasA stored 691 mg/L of fatty acids, primarily as phospholipids, inside the inner membrane after 72-h cultivation. In addition, 19% of the total CoA pool was occupied by malonyl-CoA. CONCLUSIONS Increased malonyl-CoA significantly contributed to fatty acid production, and the effect was boosted by the expanded total CoA pool. Manipulation of the intracellular CoA species is effective for fatty acid production in E. coli.

中文翻译：

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外源乙酰辅酶A羧化酶和泛酸激酶在大肠杆菌中促进脂肪酸生物合成

目的 通过在大肠杆菌中引入外源性泛酸激酶 (coaA) 和乙酰辅酶 A 羧化酶 (acc)，建立一种通过增强辅酶 A (CoA) 生物合成和丙二酰辅酶 A 供应来高效生产脂肪酸的技术。结果 从谷氨酸棒杆菌中获得的 acc 的表达在大肠杆菌中积累了 2.2 倍以上的脂肪酸。添加来自恶臭假单胞菌的 coaA 或来自谷氨酸棒杆菌的脂肪酸合酶 (fasA) 导致分别表达 acc 和 coaA 或 acc 和 fasA 的大肠杆菌细胞中脂肪酸合成增加 3.1 倍和 3.6 倍. 同时拥有所有三个基因的转化体产生的脂肪酸增加了 5.6 倍。具有 acc、coaA 和 fasA 的菌株储存了 691 mg/L 的脂肪酸，主要是磷脂，培养 72 小时后内膜内。此外，总 CoA 库的 19% 被丙二酰辅酶 A 占据。结论 丙二酰辅酶A的增加显着促进了脂肪酸的产生，并且这种效应因总辅酶A池的扩大而增强。细胞内 CoA 种类的操纵对于大肠杆菌中的脂肪酸生产是有效的。