Journal
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
Volume 108, Issue 5, Pages 359-364Publisher
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2009.05.002
Keywords
Sake yeast; Industrial diploid yeast; Alcohol acetyltransferase; Loss of heterozygosity; SED1 promoter; HELOH method
Funding
- New Bio-industry Initiatives of the Bio-oriented Technology Research Advancement Institution
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By application of the high-efficiency loss of heterozygosity (HELOH) method for disrupting genes in diploid sake yeast (Kotaka et al., Appl. Microbiol. Biotechnol., 82, 387-395 (2009)), we constructed, from a beterozygous integrant, a homozygous diploid that overexpresses the alcohol acetyltransferase gene ATF2 from the SEDI promoter, without the need for sporulation and mating. Under the conditions of sake brewing, the homozygous integrant produced 1.4 times more isoamyl acetate than the parental, heterozygous strain. Furthermore, the homozygous integrant was more genetically stable than the heterozygous recombinant. Thus, the HELOH method can produce homozygous, recombinant sake yeast that is ready to be grown on an industrial scale using the well-established procedures of sake brewing. The HELOH method, therefore, facilitates genetic modification of this rarely sporulating diploid yeast strain while maintaining those characteristics required for industrial applications. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.
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