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Secretion of mouse α‐amylase from fission yeast Schizosaccharomyces pombe : Presence of chymostatin‐sensitive protease activity in the culture medium
Author(s) -
Tokunaga Masao,
Kawamura Akiko,
Yonekyu Sayuri,
Kishida Masao,
Hishinuma Fumio
Publication year - 1993
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320090408
Subject(s) - schizosaccharomyces pombe , biology , secretion , secretory protein , schizosaccharomyces , protease , amylase , yeast , kluyveromyces lactis , microbiology and biotechnology , biochemistry , saccharomyces cerevisiae , enzyme
We have constructed two secretion vectors for Schizosaccharomyces pombe using an SV40 promoter and the secretion signals of the pGKL killer toxin complex derived from Kluyveromyces lactis . Although indigenous secretory glycoproteins tend to accumulate in the periplasmic space of S. pombe , we have succeeded in the secretion of mouse α‐amylase into the culture medium. The efficiency of secretion, processing pattern, stability and culture conditions for mouse α‐amylase were studied in S. pombe . The 128 kDa killer secretion signal was more effective in directing secretion of mouse α‐amylase than the 28 kDa killer secretion signal. We detected a chymostatin‐sensitive protease activity in the culture medium of S. pombe , which digests mouse α‐amylase secreted into the culture medium. The addition of 5 μg/ml chymostatin was shown to protect mouse α‐amylases from this degradation.