Increased RNA production in Saccharomyces cerevisiae by simultaneously overexpressing FHL1 , IFH1 , and SSF2 and deletin
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APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGY
Increased RNA production in Saccharomyces cerevisiae by simultaneously overexpressing FHL1, IFH1, and SSF2 and deleting HRP1 Xuewu Guo 1,2,3,4 & Bin Zhao 1,2 & Xinran Zhou 1,2 & Xiaofeng Ni 1,2 & Dongxia Lu 1,2 & Tingli Chen 1 & Yefu Chen 1,2,3 & Dongguang Xiao 1,2,3 Received: 5 March 2020 / Revised: 1 July 2020 / Accepted: 13 July 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Ribonucleic acid (RNA) and its degradation products are widely used in the food industry. In this study, we constructed Saccharomyces cerevisiae mutants with FHL1, IFH1, SSF1, and SSF2 overexpression and HRP1 deletion, individually to evaluate the effect on RNA production. The RNA content of recombinant strains W303-1a-FHL1, W303-1a-SSF2, and W303-1a-ΔHRP1 was increased by 14.94%, 24.4%, and 19.36%, respectively, compared with the RNA content of the parent strain. However, W303-1a-IFH1 and W303-1a-SSF1 showed no significant change in RNA production compared with the parent strain. IFH1 and FHL1 encode Ifh1p and Fhl1p, respectively, which combine to form a complex that plays a key role in the transcription of the ribosomal protein (RP) gene. Ssf2p, encoded by SSF2, plays an important role in ribosome biosynthesis and Hrp1p is a negative regulator of cell growth in S. cerevisiae. Subsequently, a high RNA production strain, W112, was constructed by simultaneously overexpressing FHL1, IFH1, and SSF2 and deleting HRP1. The RNA content of W112 was 38.8% higher than the parent strain. The growth performance, RP transcription levels, and rRNA content were also investigated in the recombinant strains. This study provides a new strategy for the construction of S. cerevisiae strains containing large amounts of RNA, and it will make a significant contribution to progress in the nucleic acid industry. Key Points • Simultaneously overexpressing FHL1, IFH1, and SSF2 and deleting HRP1 can significantly increases RNA production. • The production of RNA increased by 38.8% in Saccharomyces cerevisiae. • The cell size and growth rate of the strains with higher RNA content also increased. Keywords RNA content . Saccharomyces cerevisiae . FHL1 . IFH1 . SSF2 . HRP1
* Xuewu Guo [email protected] 1
Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Lab, College of Biotechnology of Tianjin University of Science and Technology, Tianjin 300547, China
2
Tianjin Food Safety & Low Carbon Manufacturing Collaborative Innovation Center, Tianjin 300457, China
3
Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control, Tianjin 300457, China
4
Department of Fermentation Engineering, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China
Introduction Ribonucleic acid (RNA), an important component of organisms, has many applications in various fields, such as genetic engineering, medicine, food, and agriculture (Barbalat et al. 2011; Nagodawithana 1992). For
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