Guo Can, Bin Zhiqiang, Zhang Pengjie, Tang Jing, Wang Lianqing, Chen Yefu, Xiao Dongguang, Guo Xuewu
Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education, Tianjin Industrial Microbiology Key Lab, College of Biotechnology of Tianjin University of Science and Technology, Tianjin 300457, China; Liquor Making Biological Technology and Application of Key Laboratory of Sichuan Province, Sichuan, China.
Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education, Tianjin Industrial Microbiology Key Lab, College of Biotechnology of Tianjin University of Science and Technology, Tianjin 300457, China; Liquor Making Biological Technology and Application of Key Laboratory of Sichuan Province, Sichuan, China; Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control, Tianjin 300457, China.
J Biotechnol. 2025 Feb;398:66-75. doi: 10.1016/j.jbiotec.2024.11.021. Epub 2024 Dec 3.
RNA (Ribonucleic Acid) is an essential component of organisms and is widely used in the food and pharmaceutical industries. Saccharomyces cerevisiae, recognized as a safe strain, is widely used for RNA production. In this study, the S. cerevisiae W303-1a was used as a starting strain and molecular modifications were made to the functional ncRNA-SRG1 to evaluate the effect on RNA production. At the same time, its transcriptionally associated helper genes (Spt2, Spt6 and Cha4) were overexpressed and the culture medium was supplemented with serine to induce SRG1 transcription, to increase SRG1 transcription levels and investigate its effect on intracellular RNA levels. The results showed that the intracellular RNA content of the recombinant strain W303-1a-SRG1 was 10.27 %, an increase of 11.15 % compared to the starting strain (W303-1a, with an intracellular RNA content of 9.24 %). On this basis, a gene co-overexpression strain-W303-1a-SRG1-Spt6 was constructed. Simultaneously, the addition of 2 % serine strategy was used to increase the transcription level of SRG1 and RNA content of the recombinant strain. The intracellular RNA of the recombinant strain reached 11.41 %, an increase of 23.38 % compared to the starting strain (W303-1a, without serine supplementation). In addition, the growth performance of the strain was assessed by measuring the SRG1 transcription level in the strain and plotting the growth curve. Therefore, we found that improving the transcription level of ncRNA can be used as a new idea to construct S. cerevisiae with high RNA content, which provides a strong help for subsequent research in related fields. This work provides a new strategy for increasing the nucleic acid content of S. cerevisiae.
核糖核酸（RNA）是生物体的重要组成部分，在食品和制药行业有着广泛应用。酿酒酵母被公认为安全菌株，广泛用于RNA生产。本研究以酿酒酵母W303-1a为出发菌株，对功能性非编码RNA-SRG1进行分子改造，以评估其对RNA生产的影响。同时，过表达其转录相关辅助基因（Spt2、Spt6和Cha4），并在培养基中添加丝氨酸以诱导SRG1转录，提高SRG1转录水平，研究其对细胞内RNA水平的影响。结果表明，重组菌株W303-1a-SRG1的细胞内RNA含量为10.27%，与出发菌株（W303-1a，细胞内RNA含量为9.24%）相比增加了11.15%。在此基础上，构建了基因共过表达菌株-W303-1a-SRG1-Spt6。同时，采用添加2%丝氨酸的策略提高重组菌株的SRG1转录水平和RNA含量。重组菌株的细胞内RNA达到11.41%，与未添加丝氨酸的出发菌株（W303-1a）相比增加了23.38%。此外，通过测定菌株中SRG1转录水平并绘制生长曲线来评估菌株的生长性能。因此，我们发现提高非编码RNA的转录水平可作为构建高RNA含量酿酒酵母的新思路，为后续相关领域的研究提供有力帮助。本工作为提高酿酒酵母核酸含量提供了新策略。
