SECRETION OF MOUSE ALPHA-AMYLASE FROM FISSION YEAST SCHIZOSACCHAROMYCES-POMBE - PRESENCE OF CHYMOSTATIN-SENSITIVE PROTEASE ACTIVITY IN THE CULTURE-MEDIUM
M. Tokunaga et al., SECRETION OF MOUSE ALPHA-AMYLASE FROM FISSION YEAST SCHIZOSACCHAROMYCES-POMBE - PRESENCE OF CHYMOSTATIN-SENSITIVE PROTEASE ACTIVITY IN THE CULTURE-MEDIUM, Yeast, 9(4), 1993, pp. 379-387
We have constructed two secretion vectors for Schizosaccharomyces pomb
e using an SV40 promoter and the secretion signals of the pGKL killer
toxin complex derived from Kluyveromyces lactis. Although indigenous s
ecretory glycoproteins tend to accumulate in the periplasmic space of
S. pombe, we have succeeded in the secretion of mouse alpha-amylase in
to the culture medium. The efficiency of secretion, processing pattern
, stability and culture conditions for mouse alpha-amylase were studie
d in S. pombe. The 128 kDa killer secretion signal was more effective
in directing secretion of mouse alpha-amylase than the 28 kDa killer s
ecretion signal. We detected a chymostatin-sensitive protease activity
in the culture medium of S. pombe, which digests mouse alpha-amylase
secreted into the culture medium. The addition of 5 mug/ml chymostatin
was shown to protect mouse alpha-amylases from this degradation.