A. A. Cherenkova, B. V. Sviridov, T. K. Dvoryanchikova, O. E. Melkina
{"title":"The Yarrowia lipolytica Ribosomal Promoter pRPL25 as a Promising Object for Metabolic Engineering of Production Strains","authors":"A. A. Cherenkova, B. V. Sviridov, T. K. Dvoryanchikova, O. E. Melkina","doi":"10.1134/S0003683824700224","DOIUrl":null,"url":null,"abstract":"<p> Promoters are crucial components of yeast production strains in metabolic engineering. Due to its high activity, the promoter of the <i>RPL25</i> gene, which encodes the L25 protein of the large ribosomal subunit, is a promising object for investigation and optimization. Here, to investigate the <i>Yarrowia lipolytica</i> pRPL25 promoter structure, a randomized library of its sequences was generated in 100 nucleotide increments. The strength of the obtained promoters was evaluated using the green fluorescent protein as a reporter. The results showed that the minimum functional length of the studied pRPL25 derivatives was 199 bp, and the optimal functional length was 497 bp. When the sequence was shortened to 400 bp, the expression level of the reporter decreased significantly. Based on the above, we propose that the 400–500 bp region of <i>pRPL25</i> comprises an upstream activating sequence (UAS). The pRPL25 promoter and its derivatives potentially can be used in bioengineering to create metabolic pathways with a precise level of gene expression. Strains constructed on their basis may be more efficient producers of target compounds compared to classical expression systems such as <i>Saccharomyces cerevisiae.</i></p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 9","pages":"1623 - 1630"},"PeriodicalIF":1.0000,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Applied Biochemistry and Microbiology","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1134/S0003683824700224","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Promoters are crucial components of yeast production strains in metabolic engineering. Due to its high activity, the promoter of the RPL25 gene, which encodes the L25 protein of the large ribosomal subunit, is a promising object for investigation and optimization. Here, to investigate the Yarrowia lipolytica pRPL25 promoter structure, a randomized library of its sequences was generated in 100 nucleotide increments. The strength of the obtained promoters was evaluated using the green fluorescent protein as a reporter. The results showed that the minimum functional length of the studied pRPL25 derivatives was 199 bp, and the optimal functional length was 497 bp. When the sequence was shortened to 400 bp, the expression level of the reporter decreased significantly. Based on the above, we propose that the 400–500 bp region of pRPL25 comprises an upstream activating sequence (UAS). The pRPL25 promoter and its derivatives potentially can be used in bioengineering to create metabolic pathways with a precise level of gene expression. Strains constructed on their basis may be more efficient producers of target compounds compared to classical expression systems such as Saccharomyces cerevisiae.
期刊介绍:
Applied Biochemistry and Microbiology is an international peer reviewed journal that publishes original articles on biochemistry and microbiology that have or may have practical applications. The studies include: enzymes and mechanisms of enzymatic reactions, biosynthesis of low and high molecular physiologically active compounds; the studies of their structure and properties; biogenesis and pathways of their regulation; metabolism of producers of biologically active compounds, biocatalysis in organic synthesis, applied genetics of microorganisms, applied enzymology; protein and metabolic engineering, biochemical bases of phytoimmunity, applied aspects of biochemical and immunochemical analysis; biodegradation of xenobiotics; biosensors; biomedical research (without clinical studies). Along with experimental works, the journal publishes descriptions of novel research techniques and reviews on selected topics.