Heterologous expression and efficient ethanol production of a Rhizopus glucoamylase gene in Saccharomyces cerevisiae.
Glucoamylases are inverting exo-acting starch hydrolases releasing β-glucose from the non-reducing ends of starch and related substrates. Due to the absence of glucoamylase in Saccharomyces cerevisiae, it is not capable of utilizing starch directly as energy sources without enzymatic or chemical hydrolysis for its ethanol production. In this study, we heterologously expressed a previously isolated Rhizopus arrhizus glucoamylase gene in S. cerevisiae host. The expressed glucoamylase enzyme was secreted into the culture supernatant and exhibited a molecular weight of 68 kDa on SDS-PAGE gel and western blot. In the flask ferment experiment of S. cerevisiae growing on raw starch, the RaGA transformed strains could utilize starch as energy source to produce ethanol up to a final concentration as 5%.
School of Agriculture and Bioengineering, Tianjin University, Tianjin, 300072, People's Republic of China.
Shaohui
S
Yang
Naibing
N
Jia
Minggang
M
Li
Jiehua
J
Wang
D001709:Biotechnology / Q000379:methods*; D000431:Ethanol / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics*; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics*; D006868:Hydrolysis; D011995:Recombination, Genetic / Q000235:genetics; D012233:Rhizopus / Q000201:enzymology* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013213:Starch / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D005656:Fungal Proteins; D000431:Ethanol; D013213:Starch; D005087:Glucan 1,4-alpha-Glucosidase