Label-free isolation of lipid-rich Saccharomyces cerevisiae mutant by high-throughput flow-mode Raman-activated cell sorting and multi-omics analysis for uncovering the mechanism of enhanced lipid accumulation

作者：Xiaotong Ji, Xixian Wang, Wenjun Zhou, Lin Chen, Tianzhong Liu, Jian Xu, Bo Ma · 发表于：Biotechnology for Biofuels and Bioproducts · 年份：2025 · DOI：10.1186/s13068-025-02677-8 · 被引用次数：6 · 研究领域：Microbial Metabolic Engineering and Bioproduction、Lipid metabolism and biosynthesis、Metabolomics and Mass Spectrometry Studies

BACKGROUND: Palmitoleic acid, a valuable functional fatty acid, is notably scarce in traditional oil crops, with the exception of certain wild plants such as macadamia nuts and sea buckthorn. Recently, the lipid from Saccharomyces cerevisiae was found to contain approximately 50% palmitoleic acid. Consequently, S. cerevisiae has the potential to sustainably produce palmitoleic acid through fermentation, provided that the issue of promoting its lipid content is addressed. RESULTS: In this work, based on the previously isolated oleaginous wild strain of S. cerevisiae, the mutagenesis by zeocin combined with ARTP was carried out to generate S. cerevisiae mutants, and then the high lipid content mutants were isolated using the flow-mode Raman-activated cell sorting (FlowRACS) technique, which allowed for the high-throughput selection of these mutants in a label-free and non-invasive manner. The mutant MU2R48 was finally obtained and its lipid content was 40.26%, 30.85% higher than the original type. Transcriptome and targeted metabolome analysis revealed a coordinated interaction of fatty acid precursor biosynthesis, the pentose phosphate pathway, ethanol degradation, and amino acid metabolism, synergistically channeling carbon flux from acetyl-CoA and NADPH into lipid biosynthesis. Additionally, key transcriptional regulators within the lipid metabolism network were implicated in this enhanced lipid accumulation. CONCLUSION: In this study, a mutant strain of Saccharomyces cerevi...