Wang Li, Wang Leilei, Wang Ruiming, Wang Zhaoyun, Wang Junqing, Yuan Haibo, Su Jing, Li Yan, Yang Suzhen, Han Tingting
State Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology, Jinan, Shandong 250353, China.
Key Laboratory of Shandong Microbial Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan, Shandong 250353, China.
ACS Omega. 2022 May 19;7(21):17774-17783. doi: 10.1021/acsomega.2c00972. eCollection 2022 May 31.
10-Hydroxy-2-decenoic acid (10-HDA) is an α,β-unsaturated medium-chain carboxylic acid containing a terminal hydroxyl group. It has various unique properties and great economic value. We improved the two-step biosynthesis method of 10-HDA. The conversion rate of the intermediate product -2-decenoic acid in the first step of 10-HDA synthesis could reach 93.1 ± 1.3% by combining transporter overexpression and permeation technology strategies. Moreover, the extracellular -2-decenoic acid content was five times greater than the intracellular content when 2.0% (v/v) triton X-100 and 1.2% (v/v) tween-80 were each used. In the second step of 10-HDA synthesis, we regenerated NAD(P)H by overexpressing a glucose dehydrogenase with the P450 enzyme (CYP153A33/M228L-CPR) in , improving the catalytic performance of the -2-decenoic acid terminal hydroxylation. Finally, the yield of 10-HDA was 486.5 mg/L using decanoic acid as the substrate with two-step continuous biosynthesis. Our research provides a simplified production strategy to promote the two-step continuous whole-cell catalytic biosynthesis of 10-HDA and other α,β-unsaturated carboxylic acid derivatives.
10-羟基-2-癸烯酸（10-HDA）是一种含有末端羟基的α,β-不饱和中链羧酸。它具有多种独特性质和巨大的经济价值。我们改进了10-HDA的两步生物合成方法。通过结合转运蛋白过表达和渗透技术策略，10-HDA合成第一步中中间产物-2-癸烯酸的转化率可达93.1±1.3%。此外，当分别使用2.0%（v/v）的 Triton X-100和1.2%（v/v）的吐温-80时，细胞外-2-癸烯酸含量比细胞内含量高五倍。在10-HDA合成的第二步中，我们通过在中过表达一种葡萄糖脱氢酶与P450酶（CYP153A33/M228L-CPR）来再生NAD(P)H，提高了-2-癸烯酸末端羟基化的催化性能。最后，以癸酸为底物通过两步连续生物合成，10-HDA的产量为486.5 mg/L。我们的研究提供了一种简化的生产策略，以促进10-HDA和其他α,β-不饱和羧酸衍生物的两步连续全细胞催化生物合成。
