Improvement of 2,3-butanediol production by dCas9 gene expression system in Saccharomyces cerevisiae
Saccharomyces cerevisiae has been widely used in bioproduction. To produce a target product other than ethanol, ethanol production must be decreased to enhance target production. An ethanol non-producing yeast strain was previously constructed by knocking out pyruvate decarboxylase (PDC) genes in th...
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| Veröffentlicht in: | Journal of bioscience and bioengineering 2022-03, Vol.133 (3), p.208-212 |
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| creator | Morita, Keisuke Seike, Taisuke Ishii, Jun Matsuda, Fumio Shimizu, Hiroshi |
| description | Saccharomyces cerevisiae has been widely used in bioproduction. To produce a target product other than ethanol, ethanol production must be decreased to enhance target production. An ethanol non-producing yeast strain was previously constructed by knocking out pyruvate decarboxylase (PDC) genes in the ethanol synthetic pathway. However, glucose uptake by the ethanol-non-producing yeast strain was significantly decreased. In this study, dead Cas9 (dCas9) was used to reduce ethanol synthesis during 2,3-butanediol production without reduction of glucose. The binding site of guide RNA used to effectively suppress PDC1 promoter-driven red fluorescent protein expression by dCas9 was identified and applied to control PDC1 expression. The production of 2,3-butanediol rather than ethanol was improved in repetitive test tube culture. Additionally, ethanol production was decreased and 2,3-butanediol production was increased in the strain expressing dCas9 targeting the PDC1 promoter in the third round of cultivation, compared with the control strain. |
| doi_str_mv | 10.1016/j.jbiosc.2021.12.007 |
| format | Article |
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To produce a target product other than ethanol, ethanol production must be decreased to enhance target production. An ethanol non-producing yeast strain was previously constructed by knocking out pyruvate decarboxylase (PDC) genes in the ethanol synthetic pathway. However, glucose uptake by the ethanol-non-producing yeast strain was significantly decreased. In this study, dead Cas9 (dCas9) was used to reduce ethanol synthesis during 2,3-butanediol production without reduction of glucose. The binding site of guide RNA used to effectively suppress PDC1 promoter-driven red fluorescent protein expression by dCas9 was identified and applied to control PDC1 expression. The production of 2,3-butanediol rather than ethanol was improved in repetitive test tube culture. Additionally, ethanol production was decreased and 2,3-butanediol production was increased in the strain expressing dCas9 targeting the PDC1 promoter in the third round of cultivation, compared with the control strain.</description><identifier>ISSN: 1389-1723</identifier><identifier>EISSN: 1347-4421</identifier><identifier>DOI: 10.1016/j.jbiosc.2021.12.007</identifier><identifier>PMID: 34998687</identifier><language>eng</language><publisher>Japan: Elsevier B.V</publisher><subject>2,3-Butanediol production ; Butylene Glycols - metabolism ; CRISPR interference ; dCas9 expression system ; Gene Expression ; Metabolic engineering ; Pyruvate Decarboxylase - genetics ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae - metabolism ; Saccharomyces cerevisiae Proteins - genetics ; Saccharomyces cerevisiae Proteins - metabolism</subject><ispartof>Journal of bioscience and bioengineering, 2022-03, Vol.133 (3), p.208-212</ispartof><rights>2021 The Society for Biotechnology, Japan</rights><rights>Copyright © 2021 The Society for Biotechnology, Japan. 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| ispartof | Journal of bioscience and bioengineering, 2022-03, Vol.133 (3), p.208-212 |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | 2,3-Butanediol production Butylene Glycols - metabolism CRISPR interference dCas9 expression system Gene Expression Metabolic engineering Pyruvate Decarboxylase - genetics Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism |
| title | Improvement of 2,3-butanediol production by dCas9 gene expression system in Saccharomyces cerevisiae |
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