Expression and secretion of alpha-amylase and glucoamylase in Saccharomyces cerevisiae.
alpha-Amylase genes of Bacillus licheniformis and glucoamylase cDNA of Aspergillus niger were ligated to a E. coli-yeast shuttle vector. The resultant plasmid was used to transform Saccharomyces cerevisiae to construct starch-degrading yeast strain. The results of enzyme activity assay and enzyme property analysis show that alpha-amylase and glucoamylase genes have been expressed simultaneously in yeast under the control of promoters and terminators of yeast MF-alpha 1 factor and PGK genes and over 99% of enzyme activities were secreted to the medium. The engineered yeast strain hydrolyses 97% of the starch (10%) in the medium after 6 days. The recombinant plasmid exists stably in yeast.
Luo
J
J
Biotechnology Research Center, Zhongshan University, Guangzhou, China.
He
M
M
Li
W
W
Zhang
T
T
D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis* / Q000235:genetics; D000072235:Mating Factor; D010455:Peptides / Q000235:genetics; D010735:Phosphoglycerate Kinase / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013213:Starch / Q000378:metabolism; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics
D001426:Bacterial Proteins; D005656:Fungal Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D000072235:Mating Factor; D013213:Starch; D010735:Phosphoglycerate Kinase; D000516:alpha-Amylases; D005087:Glucan 1,4-alpha-Glucosidase