Zhang Qian, Li Ning, Lyv Yunbin, Yu Shiqin, Zhou Jingwen
Science Center for Future Foods, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.
School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.
Synth Syst Biotechnol. 2022 Jul 2;7(4):1056-1063. doi: 10.1016/j.synbio.2022.06.008. eCollection 2022 Dec.
As a potential substitute for fatty acids, common low-cost oils could be used to produce acetyl-CoA derivatives, which meet the needs of low-cost industrial production. However, oils are hydrophobic macromolecules and cannot be directly transported into cells. In this study, caveolin was expressed in to absorb exogenous oils. The expression of caveolin fused with green fluorescent protein showed that caveolin mediated the formation of microvesicles in and the addition of 5,6-carboxyfluorescein showed that caveolae had the ability to transport exogenous substances into cells. The intracellular and extracellular triacylglycerol levels were then detected after the addition of soybean oil pre-stained with Nile Red, which proved that caveolae had the ability to absorb the exogenous oils. Lastly, caveolin for oils absorption and lipase from for oil hydrolysis were co-expressed in the naringenin-producing strain, resulting in naringenin production increasing from 222 mg/g DCW (dry cell weight) (231 mg/L) to 269 mg/g DCW (241 mg/L). These results suggested that the caveolin-mediated transporter independent oil transport system would provide a promising strategy for the transport of hydrophobic substrates.
作为脂肪酸的潜在替代品,常见的低成本油脂可用于生产乙酰辅酶A衍生物,以满足低成本工业生产的需求。然而,油脂是疏水性大分子,无法直接运输到细胞中。在本研究中,通过表达小窝蛋白来吸收外源油脂。与绿色荧光蛋白融合的小窝蛋白的表达表明,小窝蛋白在细胞中介导了微囊泡的形成,而添加5,6-羧基荧光素表明小窝具有将外源物质运输到细胞中的能力。在添加用尼罗红预染色的大豆油后,检测细胞内和细胞外甘油三酯水平,这证明小窝具有吸收外源油脂的能力。最后,在生产柚皮素的菌株中共表达用于吸收油脂的小窝蛋白和用于水解油脂的脂肪酶,导致柚皮素产量从222mg/g干细胞重量(231mg/L)增加到269mg/g干细胞重量(241mg/L)。这些结果表明,小窝蛋白介导的非依赖转运蛋白的油脂运输系统将为疏水性底物的运输提供一种有前景的策略。