Data underlying the study on Functional analysis of Saccharomyces cerevisiae FLO genes through optogenetic control

Flocculation in <em>Saccharomyces cerevisiae</em>, the calcium-dependent aggregation of yeast cells, is a critical phenotype with ecological and industrial significance. This study aimed to functionally dissect the contributions of individual <em>FLO</em> genes (<em>FLO1</em>, <em>FLO5,</em> <em>FLO9</em>, F<em>LO10</em>) to flocculation by employing an optogenetic circuit (OptoQ-AMP5) for precise, light-inducible control of gene expression. A <em>FLO</em>-null yeast strain (IMK1060) was engineered via CRISPR-Cas9 and homologous recombination, allowing the expression of individual <em>FLO</em> genes without native background interference. Each <em>FLO</em> gene was reintroduced into the <em>FLO</em>-null background under the control of OptoQ-AMP5. Upon light induction, strains expressing <em>FLO1</em>, <em>FLO5</em>, or <em>FLO10</em> demonstrated strong flocculation, with <em>FLO1</em> and <em>FLO5</em> forming large and structurally distinct aggregates. <em>FLO9</em> induced a weaker phenotype. Sugar inhibition assays revealed distinct sensitivities among flocculins, notably <em>FLO9</em>’s novel sensitivity to fructose and maltotriose. Additionally, <em>FLO</em>-induced changes in cell surface hydrophobicity were quantified, revealing that <em>FLO10</em> and <em>FLO1</em> conferred the greatest hydrophobicity, correlating with their aggregation strength. This work establishes a robust platform for investigating flocculation mechanisms in yeast with temporal precision. It highlights the phenotypic diversity encoded within the <em>FLO </em>gene family and their differential responses to environmental cues. The optogenetic system provides a valuable tool for both fundamental studies and the rational engineering of yeast strains for industrial fermentation processes requiring controlled flocculation.

酿酒酵母（Saccharomyces cerevisiae）中的絮凝，即酵母细胞的钙依赖性聚集，是一类兼具生态学与工业应用价值的关键表型。本研究旨在借助光遗传回路OptoQ-AMP5实现基因表达的精准光诱导调控，从功能层面解析单个FLO基因（FLO1、FLO5、FLO9、FLO10）对絮凝过程的贡献。研究人员通过CRISPR-Cas9与同源重组技术构建了FLO基因全缺失酵母菌株IMK1060，该菌株可在无内源背景干扰的条件下单独表达某一FLO基因。将每个FLO基因置于OptoQ-AMP5的调控下，重新导入至该FLO缺失菌株背景中。光诱导处理后，表达FLO1、FLO5或FLO10的菌株均呈现出显著的絮凝现象，其中FLO1与FLO5可形成体积庞大且结构特异的细胞聚集体，而FLO9诱导的絮凝表型相对较弱。糖抑制实验结果显示，不同絮凝素的糖敏感性存在显著差异，尤为突出的是FLO9对果糖与麦芽三糖展现出全新的敏感性。此外，本研究还定量测定了FLO基因诱导的细胞表面疏水性变化，结果表明FLO10与FLO1可赋予细胞最高的表面疏水性，该结果与其细胞聚集强度呈显著正相关。本研究搭建了一套具备时间精准调控能力的稳健研究平台，用于解析酵母絮凝的分子机制；同时揭示了FLO基因家族所编码的表型多样性，以及它们对环境信号的差异化响应。该光遗传系统可为基础研究以及针对需可控絮凝的工业发酵过程的酵母菌株理性改造提供极具价值的研究工具。