Proteolytic stability of recombinant human serum albumin secreted in the yeast Saccharomyces cerevisiae

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Title
Proteolytic stability of recombinant human serum albumin secreted in the yeast Saccharomyces cerevisiae
Author(s)
Kang Hyun Ah; Eui Sung Choi; W K Hong; J Y Kim; S M Ko; Jung Hoon Sohn; Sang Ki Rhee
Bibliographic Citation
Applied Microbiology and Biotechnology, vol. 53, no. 5, pp. 575-582
Publication Year
2000
Abstract
In order to direct the persistent expression of recombinant human serum albumin (HSA) from the GAL1O promoter in the yeast Saccharomyces cerevisiae, we carried out periodic feeding of galactose during shake-flask cultures. Unexpectedly, the recombinant protein secreted was observed to undergo rapid degradation, which was apparently accelerated by carbon-source feeding. The extracellular degradation of HSA occurred even in the strain deficient in the major vacuolar proteases PrA and PrB, and in the strain lacking the acidic protease Yap3p (involved in the generation of HSA-truncated fragments). Interestingly, the degradation correlated closely with the acidification of extracellular pH and thus was significantly overcome either by buffering the culture medium above pH 5.0 or by adding amino acid-rich supplements to the culture medium, which could prevent the acidification of medium pH during cultivation. Addition of arginine or ammonium salt also substantially minimized the degradation of HSA, even without buffering. The extracellular degradation activity was not detected in the cell-free culture supernatant but was found to be associated with intact cells. The results of the present study strongly suggest that the HSA secreted in S. cerevisiae is highly susceptible to the pH-dependent proteolysis mediated by cell-bound protease(s) whose activity and expression are greatly affected by the composition of the medium.
ISSN
0175-7598
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s002530051659
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > BioProcess Engineering Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
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