Secretion of calf chymosin from the filamentous fungus Aspergillus oryzae.
Active calf chymosin was secreted from Aspergillus oryzae transformants when the chymosin cDNA was expressed under the control of glucoamylase gene (glaA) promoter. Secreted prochymosin was autocatalytically activated to the chymosin (0.07-0.16 mg/l). Western blot analysis showed that a secreted protein immunoreactive with an anti-chymosin antibody was of similar size to authentic chymosin. Northern blot analysis revealed that mRNA of the chymosin cDNA was expressed at as high level as that of the glaA gene. The size and the level of the transcript were different among transformants, due to the integration position of the plasmid on the chromosome.
Tsuchiya
K
K
Research Institute of Brewing Resources Co. Ltd., Tokyo, Japan.
Gomi
K
K
Kitamoto
K
K
Kumagai
C
C
Tamura
G
G
D000818:Animals; D001236:Aspergillus oryzae / Q000378:metabolism*; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002417:Cattle; D012085:Chymosin / Q000096:biosynthesis / Q000378:metabolism*; D003001:Cloning, Molecular; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics
D011993:Recombinant Fusion Proteins; D005087:Glucan 1,4-alpha-Glucosidase; D012085:Chymosin