{"title":"Metabolic Engineering of the Yeast Schizosaccharomyces pombe Tolerant to Low pH for L-Lactic Acid Production","authors":"E. O. Anisimova, M. G. Tarutina, S. P. Sineoky","doi":"10.1134/S0003683824700133","DOIUrl":null,"url":null,"abstract":"<p> The recombinant strain based on the acidophilic yeast <i>Schizosaccharomyces pombe</i> with a cloned gene of heterologous lactate dehydrogenase (<i>LDH</i>) was used to optimize the biosynthesis of L-lactic acid. For this purpose, the effect of inactivation of pyruvate decarboxylase genes (<i>PDC</i>) on the synthesis of acetate, pyruvate, and ethanol (the main by-product in the synthesis of lactic acid) was studied. Using the wild <i>S. pombe</i> strain, we showed that the Δ<i>pdc3</i> and Δ<i>pdc4</i> deletions did not affect these indicators, while in the Δ<i>pdc1</i> mutant, ethanol biosynthesis was reduced and acetate biosynthesis was increased, and the Δ<i>pdc2</i> mutant accumulated pyruvate. The effect of deletions of the <i>PDC1</i> and <i>PDC2</i> genes on lactic acid biosynthesis was tested on a model <i>S. pombe</i> strain containing a heterologous <i>LDH</i> gene from <i>Lactobacillus pentosus.</i> It was shown that in strains with the inactivated <i>PDC2</i> gene the accumulated pyruvate was poorly consumed for the biosynthesis of lactic acid even in the presence of two different recombinant lactate dehydrogenases; the introduction of the third gene of heterologous lactate dehydrogenase led to loss of viability of the strain. At the same time, in strains with the deleted <i>PDC1</i> gene, the biosynthesis of lactic acid was enhanced with the introduction of additional <i>LDH</i> genes. The results obtained can be used in the design of industrial lactic acid producing strains.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 8","pages":"1534 - 1542"},"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/S0003683824700133","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
The recombinant strain based on the acidophilic yeast Schizosaccharomyces pombe with a cloned gene of heterologous lactate dehydrogenase (LDH) was used to optimize the biosynthesis of L-lactic acid. For this purpose, the effect of inactivation of pyruvate decarboxylase genes (PDC) on the synthesis of acetate, pyruvate, and ethanol (the main by-product in the synthesis of lactic acid) was studied. Using the wild S. pombe strain, we showed that the Δpdc3 and Δpdc4 deletions did not affect these indicators, while in the Δpdc1 mutant, ethanol biosynthesis was reduced and acetate biosynthesis was increased, and the Δpdc2 mutant accumulated pyruvate. The effect of deletions of the PDC1 and PDC2 genes on lactic acid biosynthesis was tested on a model S. pombe strain containing a heterologous LDH gene from Lactobacillus pentosus. It was shown that in strains with the inactivated PDC2 gene the accumulated pyruvate was poorly consumed for the biosynthesis of lactic acid even in the presence of two different recombinant lactate dehydrogenases; the introduction of the third gene of heterologous lactate dehydrogenase led to loss of viability of the strain. At the same time, in strains with the deleted PDC1 gene, the biosynthesis of lactic acid was enhanced with the introduction of additional LDH genes. The results obtained can be used in the design of industrial lactic acid producing strains.
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.