Expression and secretion of glucoamylase of Aspergillus niger in Saccharomyces cerevisiae.
Glucoamylase cDNA synthesized from A. niger mutant T21 was modified at 5' and 3' ends in order to clone it into yeast shuttle plasmid YFD18 and to cause fusion between cDNA and the leader region of the yeast-mating pheromone alpha-factor. The modified cDNA was then inserted into YFD18 at HindIII site. Saccharomyces cerevisiae Y33 was transformed with the resultant recombinant plasmid YFD18HH6. Analysis of transformants including halo formation on starch medium plate, SDS-PAGE of culture filtration and determination of glucoamylase activity showed that the yeast transformed with plasmid containing glucoamylase cDNA efficiently secreted glucoamylase into the medium. This fact indicated that yeast alpha-factor was able to direct the synthesis and secretion of functional glucoamylase of Aspergillus. In addition the protolytic cleavage site involved in the maturation of glucoamylase in A. niger also worked in S. cerevisiae.
Tang
G
G
Institute of Microbiology, Chinese Academy of Sciences, Beijing.
Gong
H
H
Zhong
L
L
Yang
K
K
D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics*; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D014162:Transfection
D018076:DNA, Complementary; D004271:DNA, Fungal; D005087:Glucan 1,4-alpha-Glucosidase