7-Dehydrocholesterol (7-DHC) is widely present in various organisms and is an important precursor of vitamin D. Despite significant improvements in the biosynthesis of 7-DHC, it remains insufficient to meet the industrial demands. In this study, we reported high-level production of 7-DHC in an industrial leveraging subcellular organelles. Initially, the copy numbers of were increased in combination with sterol transcriptional factor engineering and rebalanced the redox power of the strain. Subsequently, the effects of compartmentalizing the post-squalene pathway in peroxisomes were validated by assembling various pathway modules in this organelle. Furthermore, several peroxisomes engineering was conducted to enhance the production of 7-DHC. Utilizing the peroxisome as a vessel for partial post-squalene pathways, the potential of yeast for 7-dehydrocholesterol production was demonstrated by achieving a 26-fold increase over the initial production level. 7-DHC titer reached 640.77 mg/L in shake flasks and 4.28 g/L in a 10 L bench-top fermentor, the highest titer ever reported. The present work lays solid foundation for large-scale and cost-effective production of 7-DHC for practical applications.

中文翻译：

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改造二倍体酿酒酵母的亚细胞器以提高 7-脱氢胆固醇的产量


7-脱氢胆固醇(7-DHC)广泛存在于多种生物体中，是维生素D的重要前体。尽管7-DHC的生物合成取得了显着进展，但仍不足以满足工业需求。在这项研究中，我们报道了利用亚细胞器在工业中高水平生产 7-DHC。最初，结合甾醇转录因子工程增加了拷贝数，并重新平衡了菌株的氧化还原能力。随后，通过在该细胞器中组装各种途径模块来验证在过氧化物酶体中划分后角鲨烯途径的效果。此外，还进行了一些过氧化物酶体工程来提高 7-DHC 的产量。利用过氧化物酶体作为部分角鲨烯后途径的容器，酵母生产 7-脱氢胆固醇的潜力得到了证实，产量比初始水平提高了 26 倍。 7-DHC 滴度在摇瓶中达到 640.77 mg/L，在 10 L 台式发酵罐中达到 4.28 g/L，这是迄今为止报道的最高滴度。目前的工作为大规模、经济高效地生产 7-DHC 并应用于实际应用奠定了坚实的基础。