Wang Dingkang, Hao Liying, Jiao Xue, Que Zhiluo, Huang Jun, Jin Yao, Zhou Rongqing, Wang Zhonghui, Wu Chongde
College of Biomass Science and Engineering, Sichuan University, Chengdu, China.
Global Health Institute, School of Public Health, Xi'an Jiaotong University Health Science Center, Xi'an Jiaotong University, Xian, China.
J Sci Food Agric. 2024 Mar 15;104(4):2398-2405. doi: 10.1002/jsfa.13162. Epub 2023 Dec 7.
Yeast is often used to build cell factories to produce various chemicals or nutrient substances, which means the yeast has to encounter stressful environments. Previous research reported that unsaturated fatty acids were closely related to yeast stress resistance. Engineering unsaturated fatty acids may be a viable strategy for enhancing the stress resistance of cells.
In this study, two desaturase genes, OLE1 and FAD2 from Z. rouxii, were overexpressed in S. cerevisiae to determine how unsaturated fatty acids affect cellular stress tolerance of cells. After cloning and plasmid recombination, the recombinant S. cerevisiae cells were constructed. Analysis of membrane fatty acid contents revealed that the recombinant S. cerevisiae with overexpression of OLE1 and FAD2 genes contained higher levels of fatty acids C16:1 (2.77 times), C18:1 (1.51 times) and C18:2 (4.15 times) than the wild-type S. cerevisiae pY15TEF1. In addition, recombinant S. cerevisiae cells were more resistant to multiple stresses, and exhibited improved membrane functionality, including membrane fluidity and integrity.
These findings demonstrated that strengthening the expression of desaturases was beneficial to stress tolerance. Overall, this study may provide a suitable means to build a cell factory of industrial yeast cells with high tolerance during biological manufacturing. © 2023 Society of Chemical Industry.
酵母常用于构建细胞工厂以生产各种化学品或营养物质,这意味着酵母必须应对压力环境。先前的研究报道,不饱和脂肪酸与酵母的抗逆性密切相关。改造不饱和脂肪酸可能是增强细胞抗逆性的一种可行策略。
在本研究中,将来自鲁氏接合酵母的两个去饱和酶基因OLE1和FAD2在酿酒酵母中过表达,以确定不饱和脂肪酸如何影响细胞的应激耐受性。经过克隆和质粒重组后,构建了重组酿酒酵母细胞。膜脂肪酸含量分析表明,过表达OLE1和FAD2基因的重组酿酒酵母中,脂肪酸C16:1(2.77倍)、C18:1(1.51倍)和C18:2(4.15倍)的含量高于野生型酿酒酵母pY15TEF1。此外,重组酿酒酵母细胞对多种胁迫更具抗性,并表现出改善的膜功能,包括膜流动性和完整性。
这些发现表明,增强去饱和酶的表达有利于提高抗逆性。总体而言,本研究可能为构建在生物制造过程中具有高耐受性的工业酵母细胞工厂提供一种合适的方法。© 2023化学工业协会。