Pathway Refactoring for Efficient 7-Dehydrocholesterol Production in Saccharomyces cerevisiae.

IF 3.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

ACS Synthetic Biology Pub Date : 2025-07-18 Epub Date: 2025-06-27 DOI:10.1021/acssynbio.5c00032

Yuchen Han, Huayi Gao, Shuo Wang, Haiyan Ju, Ran Ge, Chaoyou Xue, Weidong Liu, Xiaolong Jiang, Wuyuan Zhang

{"title":"Pathway Refactoring for Efficient 7-Dehydrocholesterol Production in Saccharomyces cerevisiae.","authors":"Yuchen Han, Huayi Gao, Shuo Wang, Haiyan Ju, Ran Ge, Chaoyou Xue, Weidong Liu, Xiaolong Jiang, Wuyuan Zhang","doi":"10.1021/acssynbio.5c00032","DOIUrl":null,"url":null,"abstract":"7-Dehydrocholesterol (7-DHC) is a subcutaneous sterol and a precursor to various active vitamin D3. Here, a Saccharomyces cerevisiae strain equipped with the de novo biosynthetic pathway for 7-DHC was constructed. 109.0 mg L-1 of 7-DHC was achieved initially by introducing heterologous 24-dehydrocholesterol reductase (DHCR24) and overexpressing vital enzymes. Following these modifications, the dynamic regulation of the ergosterol pathway and multicopy expression of DHCR24 resulted in an 86.3% increase in the 7-DHC titer. Subsequently, the effects of several organic solvents and surfactants on 7-DHC production were also explored. The addition of ε-polylysine increased the titer of 7-DHC by 99.1%. Finally, by assembling the pathway in peroxisomes and rebalancing the redox levels, the 7-DHC titer reached 517.4 mg L-1 in shake flasks. Scale-up fermentation with a 5 L bioreactor demonstrated that 3.26 g L-1 of 7-DHC was produced. The pathway refactoring strategy provides efficient production of 7-DHC in a sustainable manner.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":"2609-2618"},"PeriodicalIF":3.7000,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Synthetic Biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1021/acssynbio.5c00032","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/6/27 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

引用次数: 0

Abstract

7-Dehydrocholesterol (7-DHC) is a subcutaneous sterol and a precursor to various active vitamin D₃. Here, a Saccharomyces cerevisiae strain equipped with the de novo biosynthetic pathway for 7-DHC was constructed. 109.0 mg L^-1 of 7-DHC was achieved initially by introducing heterologous 24-dehydrocholesterol reductase (DHCR24) and overexpressing vital enzymes. Following these modifications, the dynamic regulation of the ergosterol pathway and multicopy expression of DHCR24 resulted in an 86.3% increase in the 7-DHC titer. Subsequently, the effects of several organic solvents and surfactants on 7-DHC production were also explored. The addition of ε-polylysine increased the titer of 7-DHC by 99.1%. Finally, by assembling the pathway in peroxisomes and rebalancing the redox levels, the 7-DHC titer reached 517.4 mg L^-1 in shake flasks. Scale-up fermentation with a 5 L bioreactor demonstrated that 3.26 g L^-1 of 7-DHC was produced. The pathway refactoring strategy provides efficient production of 7-DHC in a sustainable manner.

查看原文本刊更多论文

酿酒酵母高效生产7-脱氢胆固醇的途径重构

7-脱氢胆固醇（7-DHC）是一种皮下固醇，是多种活性维生素D3的前体。本文构建了一株具有7-DHC生物合成新途径的酿酒酵母。通过引入异源24-脱氢胆固醇还原酶（DHCR24）并过表达重要酶，初步获得109.0 mg L-1的7-DHC。经过这些修饰，麦角甾醇通路的动态调控和DHCR24的多拷贝表达导致7-DHC滴度升高86.3%。随后，探讨了几种有机溶剂和表面活性剂对7-DHC合成的影响。ε-聚赖氨酸的添加使7-DHC的效价提高了99.1%。最后，通过在过氧化物酶体中组装途径并重新平衡氧化还原水平，在摇瓶中7-DHC滴度达到517.4 mg L-1。5 L生物反应器放大发酵，7-DHC产量为3.26 g L-1。路径重构策略以可持续的方式提供了7-DHC的高效生产。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

ACS Synthetic Biology 生物-

CiteScore

8.00

自引率

10.60%

发文量

380

审稿时长

6-12 weeks

期刊介绍： The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.