Abstract
The aspartic protease gene of a zygomycete fungus Mucor pusillus was expressed in Saccharomyces cerevisiae under the control of the yeast GAL7 promoter. A putative preproenzyme with an NH2-terminal extension of 66 amino acids directed by the gene was processed in yeast cells and the mature enzyme, whose NH2-terminus was identical to that of the Mucor enzyme, was efficiently secreted into the medium at a concentration exceeding 150 mg/l. The enzyme secreted from the recombinant yeast was more glycosylated than the native Mucor enzyme but its enzymatic properties were almost identical with those of the native enzyme, which has been used as a milk coagulant in cheese manufacture.
MeSH Terms
Amino Acid Sequence
Animals
Aspartic Acid Endopeptidases
Calcium
Cloning, Molecular
Endopeptidases/genetics,metabolism
Gene Expression Regulation
Genes, Fungal
Glycosylation
Milk
Molecular Sequence Data
Mucor/enzymology,genetics
Plasmids
Protein Processing, Post-Translational
Recombinant Proteins/metabolism
Saccharomyces cerevisiae/genetics
Chemicals
Recombinant Proteins
Endopeptidases
Aspartic Acid Endopeptidases
rennin-like enzyme (Aspergillus ochraceus)
Calcium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Yamashita T
Department of Agricultural Chemistry, Faculty of Agriculture, University of Tokyo, Japan.
Tonouchi N
Uozumi T
Beppu T
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