通过在大肠杆菌中调控不饱和脂肪酸生物合成来利用多种原料生产中链长度聚羟基烷酸。
Producing medium-chain-length polyhydroxyalkanoate from diverse feedstocks by deregulating unsaturated fatty acid biosynthesis in Escherichia coli.
机构信息
Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan.
Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan; Innovation and Development Center of Sustainable Agriculture, National Chung Hsing University, Taichung 402, Taiwan.
出版信息
Bioresour Technol. 2022 Dec;365:128078. doi: 10.1016/j.biortech.2022.128078. Epub 2022 Oct 8.
The fatty acid metabolism in Escherichia coli has served as a basic metabolic chassis for medium-chain-length polyhydroxyalkanoate (mcl-PHA) production. In this study, the phaG and phaC1 genes from Pseudomonas entomophila L48 were first cloned as pGRN08. E. coli BL21P (E. coli BL21(DE3) ΔptsG) containing pGRN08 was able to produce 23 ± 3 and 7 ± 0 mg/L homopolymer poly(3-hydroxydecanoate)(P(3HD)) from glucose and xylose, respectively. Next, a gene, PSEEN0908 (encoding a putative 3-hydroxyacyl-CoA ligase), from P. entomophila L48 was found to increase the performance of mcl-PHA production. The induction of the fatty acid biosynthesis repressor (FabR), a transcription regulator that represses UFA biosynthesis, in E. coli substantially increased the mcl-PHA production by an order of magnitude from both unrelated and related carbon source conversion. A mcl-PHA concentration of 179 ± 1 mg/L and a content of 5.79 ± 0.16 % were obtained, where 31 mol% was 3-hydroxyoctanoate (3HO) and 69 mol% was 3HD.
大肠杆菌中的脂肪酸代谢已成为中链长度多羟基烷酸（mcl-PHA）生产的基本代谢底盘。在本研究中，首先将恶臭假单胞菌 L48 的 phaG 和 phaC1 基因克隆为 pGRN08。含有 pGRN08 的大肠杆菌 BL21P（大肠杆菌 BL21(DE3)ΔptsG）能够分别从葡萄糖和木糖生产 23±3 和 7±0mg/L 的均聚物聚(3-羟基癸酸)(P(3HD))。接下来，发现恶臭假单胞菌 L48 的基因 PSEEN0908（编码一种假定的 3-羟酰基辅酶 A 连接酶）能够提高 mcl-PHA 的生产性能。脂肪酸生物合成抑制剂（FabR）的诱导，即一种抑制 UFA 生物合成的转录调节剂，可使大肠杆菌中无关和相关碳源转化的 mcl-PHA 产量提高一个数量级。获得了 179±1mg/L 的 mcl-PHA 浓度和 5.79±0.16%的含量，其中 31mol%为 3-羟基辛酸（3HO），69mol%为 3HD。
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