Jiangxi Key Laboratory of Bioprocess Engineering and Co-Innovation Center for In-Vitro Diagnostic Reagents and Devices of Jiangxi Province, College of Life Sciences, Jiangxi Science and Technology Normal University, Nanchang, 330013, China.
J Ind Microbiol Biotechnol. 2019 Feb;46(2):231-239. doi: 10.1007/s10295-018-02123-9. Epub 2019 Jan 2.
Salt stress can trigger several physiological responses in microorganisms such as the increasing accumulation of unsaturated fatty acid, which was biosynthesized by delta-9 fatty acid desaturases (D9D) at the first step. In the present study, two D9D genes, designated AoD9D1 and AoD9D2, were isolated from Aspergillus oryzae. The expression analysis showed that AoD9D1 and AoD9D2 were upregulated under salt stress. To investigate the function of AoD9D, transgenic Saccharomyces cerevisiae strains that heterologously expressed AoD9D were exposed to salinity condition. These transgenic strains exhibited greater tolerance to salt stress than wild-type strains, and the heterologous expression of AoD9D increased the content in unsaturated fatty acids as compared to control cells. Moreover, AoD9D1 and AoD9D2 both contained fatty acid desaturase (FAD) and cytochrome b5-like Heme/Steroid-binding domains (Cyt-b5). S. cerevisiae separately transformed with the gene fragments coding for the FAD and Cyt-b5 domains in the AoD9D1 protein grew better and accumulated a higher concentration of unsaturated FAs than the control. Altogether, the heterologous expression of AoD9D enhanced the tolerance of transgenic S. cerevisiae to high salinity stress with increased accumulation of unsaturated fatty acid. The results provide some practical basis for the successful development of salt-tolerant fermentation microorganisms.
盐胁迫会引发微生物的几种生理反应,例如不饱和脂肪酸的积累增加,这是由 delta-9 脂肪酸去饱和酶(D9D)在第一步合成的。在本研究中,从米曲霉中分离出了两个 D9D 基因,命名为 AoD9D1 和 AoD9D2。表达分析表明,AoD9D1 和 AoD9D2 在盐胁迫下上调表达。为了研究 AoD9D 的功能,将异源表达 AoD9D 的转基因酿酒酵母菌株暴露于盐度条件下。与野生型菌株相比,这些转基因菌株对盐胁迫的耐受性更强,并且与对照细胞相比,AoD9D 的异源表达增加了不饱和脂肪酸的含量。此外,AoD9D1 和 AoD9D2 都含有脂肪酸去饱和酶(FAD)和细胞色素 b5 样血红素/固醇结合结构域(Cyt-b5)。分别转化了编码 AoD9D1 蛋白中 FAD 和 Cyt-b5 结构域的基因片段的 S. cerevisiae 生长得更好,积累的不饱和 FA 浓度高于对照。总之,AoD9D 的异源表达增强了转基因 S. cerevisiae 对高盐胁迫的耐受性,并增加了不饱和脂肪酸的积累。该结果为成功开发耐盐发酵微生物提供了一些实际依据。