利用多模块代谢工程和外排工程提高重组酿酒酵母的番茄红素产量

IF 3.2 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Guangxi Huang, Jiarong Li, Jingyuan Lin, Changqing Duan, Guoliang Yan
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引用次数: 0

摘要

番茄红素因其抗氧化、抗癌和抗炎特性而被广泛应用于食品工业和医学领域。然而,利用底盘细胞实现番茄红素的高效工业化生产仍然是一项重大挑战。在此,我们尝试整合多种代谢工程策略,在酿酒酵母中建立高效、平衡的番茄红素生物合成系统。首先,将番茄红素合成途径模块化,依次提高甲羟戊酸途径、乙酰-CoA供应模块和番茄红素外源酶模块的代谢通量。模块化操作使乙酰-CoA高效地转化到番茄红素合成的下游途径,使番茄红素产量提高了3.1倍。其次,我们引入了醋酸作为外源碳源,并利用醋酸可抑制启动子取代天然的ERG9启动子。这种方法不仅增加了乙酰-CoA 的供应,还同时减少了竞争性麦角甾醇途径的通量。因此,番茄红素的产量进一步增加了 42.3%。第三,我们通过过表达 ABC 转运体来促进番茄红素外流,从而优化 NADPH 的供应并减轻细胞毒性。与对照菌株相比,获得的菌株 YLY-PDR11 细胞外番茄红素含量增加了 12.7 倍。最后,番茄红素总产量达到 343.7 毫克/升,是初始菌株 YLY-04 的 4.3 倍。我们的研究结果表明,将多模块代谢工程与外排工程相结合是提高番茄红素产量的有效方法。这种策略也可用于在酿酒酵母中过度生产其他具有类似合成和储存模式的理想异戊二烯化合物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Multi-modular metabolic engineering and efflux engineering for enhanced lycopene production in recombinant Saccharomyces cerevisiae
Lycopene has been widely used in the food industry and medical field due to its antioxidant, anti-cancer, and anti-inflammatory properties. However, achieving efficient manufacture of lycopene using chassis cells on an industrial scale remains a major challenge. Herein, we attempted to integrate multiple metabolic engineering strategies to establish an efficient and balanced lycopene biosynthetic system in Saccharomyces cerevisiae. First, the lycopene synthesis pathway was modularized to sequentially enhance the metabolic flux of the Mevalonate pathway, the acetyl-CoA supply module, and lycopene exogenous enzymatic module. The modular operation enabled the efficient conversion of acetyl-CoA to downstream pathway of lycopene synthesis, resulting in a 3.1-fold increase of lycopene yield. Second, we introduced acetate as an exogenous carbon source and utilized an acetate-repressible promoter to replace the natural ERG9 promoter. This approach not only enhanced the supply of acetyl-CoA but also concurrently diminished the flux towards the competitive ergosterol pathway. As a result, a further 42.3% increase in lycopene production was observed. Third, we optimized NADPH supply and mitigated cytotoxicity by overexpressing ABC transporters to promote lycopene efflux. The obtained strain YLY-PDR11 showed a 12.7-fold increase in extracellular lycopene level compared to the control strain. Finally, the total lycopene yield reached 343.7mg/L, which was 4.3 times higher than that of the initial strain YLY-04. Our results demonstrate that combining multi-modular metabolic engineering with efflux engineering is an effective approach to improve the production of lycopene. This strategy can also be applied to the overproduction of other desirable isoprenoid compounds with similar synthesis and storage patterns in S. cerevisiae.
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来源期刊
Journal of Industrial Microbiology & Biotechnology
Journal of Industrial Microbiology & Biotechnology 工程技术-生物工程与应用微生物
CiteScore
7.70
自引率
0.00%
发文量
25
审稿时长
3 months
期刊介绍: The Journal of Industrial Microbiology and Biotechnology is an international journal which publishes papers describing original research, short communications, and critical reviews in the fields of biotechnology, fermentation and cell culture, biocatalysis, environmental microbiology, natural products discovery and biosynthesis, marine natural products, metabolic engineering, genomics, bioinformatics, food microbiology, and other areas of applied microbiology
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