Construction of self-cloning, indigenous wine strains of Saccharomyces cerevisiae with enhanced glycerol and glutathione production.
To improve wine taste and flavor stability, a novel indigenous strain of Saccharomyces cerevisiae with enhanced glycerol and glutathione (GSH) production for winemaking was constructed. ALD6 encoding an aldehyde dehydrogenases of the indigenous yeast was replaced by a GPD1 and CUP1 gene cassette, which are responsible for NAD-dependent glycerol-3-phosphatase dehydrogenase and copper resistance, respectively. Furthermore, the α-acetohydroxyacid synthase gene ILV2 of the indigenous yeast was disrupted by integration of the GSH1 gene which encodes γ-glutamylcysteine synthetase and the CUP1 gene cassette. The fermentation capacity of the recombinant was similar to that of the wild-type strain, with an increase of 21 and 19 % in glycerol and GSH production. No heterologous DNA was harbored in the recombinant in this study.
College of Enology, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.
Rui-Ying
RY
Hao
Yan-Lin
YL
Liu
Zhao-Yue
ZY
Wang
Bo-Run
BR
Zhang
D000095:Acetolactate Synthase / Q000235:genetics / Q000378:metabolism; D000445:Aldehyde Oxidoreductases / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D055786:Gene Knockout Techniques; D005721:Glutamate-Cysteine Ligase / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D005990:Glycerol / Q000378:metabolism*; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+) / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014920:Wine / Q000382:microbiology*
C500769:CUP1-1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D008668:Metallothionein; C568531:GPD1 protein, S cerevisiae; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+); D000445:Aldehyde Oxidoreductases; C018699:aldehyde dehydrogenase (NAD(P)+); D000095:Acetolactate Synthase; C490862:GSH1 protein, S cerevisiae; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione; D005990:Glycerol