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Title 

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

Authors 

Kwang Hyun AhS J KimEui Sung ChoiSang Ki RheeBong Hyun Chung

Publisher 

Springer Verlag (Germany)

Issue Date 

1998

Citation 

Applied Microbiology and Biotechnology, vol. 50, no. 2, pp. 187-192

Keywords 

aspartic endopeptidasesaspartic proteinasedrug manufactureparathyroid hormonepeptide fragmentspolyacrylamide gel electrophoresisprotease inhibitorsprotein processing, post-translationalrecombinant proteinsSaccharomyces cerevisiae

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

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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