Improved production of chymosin in Aspergillus by expression as a glucoamylase-chymosin fusion.
We have extended the work on chymosin production in Aspergillus by constructing an expression vector in which the cDNA encoding bovine prochymosin B was fused in frame immediately following the codon for the last amino acid of the A. awamori glucoamylase (glaA) gene. Transformation of A. awamori with this plasmid led to the secretion of considerably higher amounts of chymosin than obtained with previous chymosin expression vectors. We present evidence that mature chymosin is autocatalytically released from the glucoamylase-chymosin fusion protein after secretion.
Genencor Inc., South San Francisco, CA 94080.
M
M
Ward
L J
LJ
Wilson
K H
KH
Kodama
M W
MW
Rey
R M
RM
Berka
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D012085:Chymosin / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004271:DNA, Fungal; D005822:Genetic Vectors / Q000235:genetics; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010957:Plasmids; D014176:Protein Biosynthesis / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D004271:DNA, Fungal; D011993:Recombinant Fusion Proteins; D005087:Glucan 1,4-alpha-Glucosidase; D012085:Chymosin