Efficient production of intact human parathyroid hormone in s Saccharomyces serevisiae mutant deficient in yeast aspartic protease 3 (YAP3)

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Title
Efficient production of intact human parathyroid hormone in s Saccharomyces serevisiae mutant deficient in yeast aspartic protease 3 (YAP3)
Author(s)
Kwang Hyun Ah; S J Kim; Eui Sung Choi; Sang Ki Rhee; Bong Hyun Chung
Bibliographic Citation
Applied Microbiology and Biotechnology, vol. 50, no. 2, pp. 187-192
Publication Year
1998
Abstract
When human parathyroid hormone (hPTH) is expressed as a secretory product in yeast, the main problem is the aberrant proteolytic cleavage that reduces tile yield of intact protein. To overcome this problem, we developed an hPTH expression system using a host strain in which the YAP3 gene encoding yeast aspartic protease 3 (YAP3) was disrupted. After 48 h of culture, most of the hPTH secreted by the yap3 disruptant remained intact, whereas more than 90% of the hPTH secreted by the wild-type strain was cleaved. When the authentic hPTH was incubated in each of the culture supernatants of untransformed yap3 disruptant and wild-type strain, the proteolysis proceeded much more slowly in the culture supernatant of yap3 disruptant than in that of the wild type. The extent of hPTH proteolysis was also significantly reduced by the addition of pepstatin A, a specific aspartic protease inhibitor. The results suggest that YAP3 is involved in the internal cleavage of hPTH expressed in yeast. The correct processing of the intact hPTH secreted in the yap3 disruptant demonstrates that the yeast mutant lacking the YAP3 activity is a suitable host for the high-level expression of intact hPTH.
ISSN
0175-7598
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s002530051275
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > BioProcess Engineering Center > 1. Journal Articles
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