Isolation of Auxotrophic Mutants of Diploid Industrial Yeast Strains after UV Mutagenesis

• Shinji Hashimoto

  Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai

• Mayumi Ogura

  Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai

• Kazuo Aritomi

  Yamaguchi Prefectural Industrial Technology Institute, Asutopia, Ube, Japan

• Hisashi Hoshida

  Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai

• Yoshinori Nishizawa

  Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai

• Rinji Akada

  Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai

抄録

Auxotrophic mutants of the yeast Saccharomyces cerevisiae are usually isolated in haploid strains because the isolation of recessive mutations in diploids is thought to be difficult due to the presence of two sets of genes. We show here that auxotrophic mutants of diploid industrial sake yeast strains were routinely obtained by a standard mutant selection procedure following UV mutagenesis. We isolated His − , Met − , Lys − , Trp − , Leu − , Arg − , and Ura − auxotrophic mutants of five sake strains, Kyokai no. 7, no. 9, no. 10, no. 701, and no. 901, by screening only 1,700 to 3,400 colonies from each treated strain. Wild-type alleles were cloned and used as markers for transformation. With HIS3 as a selectable marker, the yeast TDH3 overexpression promoter was inserted upstream of ATF1 , encoding alcohol acetyltransferase, by one-step gene replacement in a his3 mutant of Kyokai no. 7. The resulting strain contained exclusively yeast DNA, making it acceptable for commercial use, and produced a larger amount of isoamyl acetate, a banana-like flavor. We argue that the generally recognized difficulty of isolating auxotrophic mutants of diploid industrial yeast strains is misleading and that genetic techniques used for haploid laboratory strains are applicable for this purpose.

収録刊行物

• Applied and Environmental Microbiology

  Applied and Environmental Microbiology 71 (1), 312-319, 2005-01

  American Society for Microbiology

キーワード

• Ecology
• Applied Microbiology and Biotechnology
• Food Science
• Biotechnology

詳細情報

• CRID: 1364233268463672576
• NII論文ID: 80017195017
• DOI: 10.1128/aem.71.1.312-319.2005
• ISSN: 10985336
• ISSN: 00992240
• Web Site: https://journals.asm.org/doi/pdf/10.1128/AEM.71.1.312-319.2005
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