The construction of a stable starch-fermenting yeast strain using genetic engineering and rare-mating.
To develop a yeast strain that is able to produce ethanol directly from starch, alpha-amylase cDNA (originated from mouse salivary glands) was introduced into the hyploid Saccharomyces diastiticus cells secreting glucoamylase by using a linearized integrating vector. The integrating vector contains a LEU2 gene and the inside of the LEU2 gene was cut by KpnI to make the linearized vector. One of the transformants exhibited 100% mitotic stability after 100 generations of cell multiplication. To improve its ethanol-fermentability, the haploid transformant was rare-mated with a polyploid industrial strain having no amylase activity. The resulting hybrid RH51 produced 7.5 (w/v) ethanol directly from 20% (w/v) soluble starch and its mitotic stability was 100% at the end of fermentation.
Kim
T G
TG
Center for Genetic Engineering Research, University of Suwon, Korea.
Kim
K
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D000818:Animals; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D003470:Culture Media; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics; D005818:Genetic Engineering; D005822:Genetic Vectors; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis / Q000235:genetics; D006238:Haploidy; D051379:Mice; D008938:Mitosis / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011123:Polyploidy; D015183:Restriction Mapping; D012440:Saccharomyces / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D012469:Salivary Glands / Q000201:enzymology; D013213:Starch / Q000737:chemistry / Q000378:metabolism*; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000096:biosynthesis / Q000235:genetics
D003470:Culture Media; D018076:DNA, Complementary; D000431:Ethanol; D013213:Starch; D000516:alpha-Amylases; D005087:Glucan 1,4-alpha-Glucosidase