Secretion of Aspergillus oryzae alkaline protease in an osmophilic yeast, Zygosaccharomyces rouxii.
To produce Aspergillus oryzae alkaline protease (Alp) in an osmophilic yeast Zygosaccharomyces rouxii, we constructed an expression plasmid consisting of the Z. rouxii glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter, the prepro-Alp cDNA of A. oryzae, the whole sequence of Z. rouxii plasmid pSR1, and the G418 resistant gene. The resulting plasmid, when introduced into Z. rouxii cells, directed the secretion of a large amount (about 300 mg/l) of Alp into the culture medium. The N-terminus and specific activity of the enzyme were identical to those of A. oryzae Alp.
Research and Development Division, Kikkoman Corporation, Chiba, Japan.
Y
Y
Ogawa
H
H
Tatsumi
S
S
Murakami
Y
Y
Ishida
K
K
Murakami
A
A
Masaki
H
H
Kawabe
H
H
Arimura
E
E
Nakano
H
H
Motai
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes; D004352:Drug Resistance, Microbial / Q000235:genetics; D004792:Enzyme Precursors / Q000235:genetics; D015870:Gene Expression; D005839:Gentamicins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics; D004718:Saccharomycetales / Q000201:enzymology* / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
D004792:Enzyme Precursors; D005839:Gentamicins; D004247:DNA; C010680:antibiotic G 418; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D004262:DNA Restriction Enzymes; D012697:Serine Endopeptidases; C026109:oryzin