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Information × Registration Number 0221U101116, (0119U001677) , R & D reports Title Study of the mechanism of action of new genes in the regulation of riboflavin synthesis in flavinogenic yeast Candida famata popup.stage_title Дослідження ролі генів SEF1 та VMA1 для надсинтезу рибофлавіну у флавіногенних дріжджів Candida famata. Head Sibirny Andriy A., Доктор біологічних наук Registration Date 15-01-2021 Organization Institute of Cell Biology of the National Academy of Sciences of Ukraine popup.description1 popup.description2 Research report: 24 pages, 4 figures, 3 tables, 35 sources. Object of research: riboflavin biosynthesis and its regulation in yeast The purpose of research: to study the molecular mechanisms of regulation of the synthesis of biologically active compounds in yeast. Research methods: genetic engineering, molecular biological, genetic, microbiological, biochemical As a result of the work: A number of yeast plasmids for the expression of the SEF1 Candida famata gene have been constructed under the control of promoters of this gene of various flavinogenic (C. famata, Candida albicans, Candida tropicalis) and non-flavinogenic (Pichia stipitis, Saccharomyces cerevisiae) yeast species. Appropriate transformants based on C. famata sef1Δ strain were obtained. By biochemical analysis, it was found that the promoters of the SEF1 gene of the flavinogenic yeast C. albicans and C. tropicalis, combined with the ARI (open reading frame) of the SEF1 gene of the C. famata restored riboflavin supersynthesis in the C. famata sef1Δ strain. Mutants with deletion of the SFU1 gene were constructed and their flavinogenic activity was studied. Deletion of the SFU1 gene has been shown to lead to a fivefold increase in riboflavin production compared to the original wild-type strain. A strain with a deletion of VMA1 based on C. famata L20105 was obtained, which accumulated 1.4 times less biomass and 9 times more riboflavin than the wild-type strain L20105. The yield of riboflavin (in terms of the amount of synthesized riboflavin per gram of dry cell mass) was 12 times higher in the deletion strain compared to the parent. The results revealed new promising targets for metabolic engineering of flavinogenic strains of C. famata to improve the production of vitamin B2. Key words: Yeast Candida famata, REGULATION OF RIBOFLAVIN PRODUCTION, TRANSCRIPTION FACTORS Sef1 and Sfu1, VACUOLAR ATPase, VMA GENE Product Description popup.authors Andreieva Yuliia A. Dmytruk Kostiantyn V. Petrovska Yana O. Faiura Liubov R. Fedorovych Dariia V. Tsyrulnyk Andrii O. popup.nrat_date 2021-01-15 Close

Information × Registration Number 0221U101116, (0119U001677) , R & D reports Title Study of the mechanism of action of new genes in the regulation of riboflavin synthesis in flavinogenic yeast Candida famata popup.stage_title Дослідження ролі генів SEF1 та VMA1 для надсинтезу рибофлавіну у флавіногенних дріжджів Candida famata. Head Sibirny Andriy A., Доктор біологічних наук Registration Date 15-01-2021 Organization Institute of Cell Biology of the National Academy of Sciences of Ukraine popup.description1 popup.description2 Research report: 24 pages, 4 figures, 3 tables, 35 sources. Object of research: riboflavin biosynthesis and its regulation in yeast The purpose of research: to study the molecular mechanisms of regulation of the synthesis of biologically active compounds in yeast. Research methods: genetic engineering, molecular biological, genetic, microbiological, biochemical As a result of the work: A number of yeast plasmids for the expression of the SEF1 Candida famata gene have been constructed under the control of promoters of this gene of various flavinogenic (C. famata, Candida albicans, Candida tropicalis) and non-flavinogenic (Pichia stipitis, Saccharomyces cerevisiae) yeast species. Appropriate transformants based on C. famata sef1Δ strain were obtained. By biochemical analysis, it was found that the promoters of the SEF1 gene of the flavinogenic yeast C. albicans and C. tropicalis, combined with the ARI (open reading frame) of the SEF1 gene of the C. famata restored riboflavin supersynthesis in the C. famata sef1Δ strain. Mutants with deletion of the SFU1 gene were constructed and their flavinogenic activity was studied. Deletion of the SFU1 gene has been shown to lead to a fivefold increase in riboflavin production compared to the original wild-type strain. A strain with a deletion of VMA1 based on C. famata L20105 was obtained, which accumulated 1.4 times less biomass and 9 times more riboflavin than the wild-type strain L20105. The yield of riboflavin (in terms of the amount of synthesized riboflavin per gram of dry cell mass) was 12 times higher in the deletion strain compared to the parent. The results revealed new promising targets for metabolic engineering of flavinogenic strains of C. famata to improve the production of vitamin B2. Key words: Yeast Candida famata, REGULATION OF RIBOFLAVIN PRODUCTION, TRANSCRIPTION FACTORS Sef1 and Sfu1, VACUOLAR ATPase, VMA GENE Product Description popup.authors Andreieva Yuliia A. Dmytruk Kostiantyn V. Petrovska Yana O. Faiura Liubov R. Fedorovych Dariia V. Tsyrulnyk Andrii O. popup.nrat_date 2021-01-15 Close