Document Detail


Multiple-yapsin-deficient mutant strains for high-level production of intact recombinant proteins in Saccharomyces cerevisiae.
MedLine Citation:
PMID:  20599573     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The yapsin family of aspartic proteases, located at cell surface, has a common specificity for paired or single basic reside cleavage sites of proproteins. Our previous study reported that the aberrant proteolytic cleavage of secretory recombinant human parathyroid hormone (hPTH) protein was problematic at late stages of fed-batch cultivations, even in the Saccharomyces cerevisiae mutant strain deficient in yapsin 1 (yps1Delta). To overcome this problem, we constructed a set of S. cerevisiae mutant strains lacking several members of the yapsin family through disruption of the YPS genes coding for yapsin 1, 2, 3, 6, and 7 proteases in various combinations. The multiple YPS-deletion mutant strains did not show detectable growth defects under normal growth conditions, although some of them were hypersensitive to hygromycin B, acid (pH 3.5) and alkali (pH 8.0) conditions. The quintuple disruptant (yps1Deltayps2Deltayps3Deltayps6Deltayps7Delta) was the most efficient in preventing the proteolytic degradation of hPTH in fed-batch cultivations. The present data strongly indicate the involvement of other yapsin members besides Yps1p in the proteolysis of secretory recombinant proteins, particularly under high-density growth conditions.
Authors:
Eun Young Cho; Seon Ah Cheon; Hyunah Kim; Jinho Choo; Dong-Jik Lee; Ho Myung Ryu; Sang Ki Rhee; Bong-Hyun Chung; Jeong-Yoon Kim; Hyun Ah Kang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-06-25
Journal Detail:
Title:  Journal of biotechnology     Volume:  149     ISSN:  1873-4863     ISO Abbreviation:  J. Biotechnol.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-11     Completed Date:  2010-11-23     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8411927     Medline TA:  J Biotechnol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1-7     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2010 Elsevier B.V. All rights reserved.
Affiliation:
Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea.
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MeSH Terms
Descriptor/Qualifier:
Aspartic Acid Endopeptidases / deficiency,  genetics,  metabolism*
Humans
Parathyroid Hormone / genetics,  metabolism
Recombinant Proteins / genetics,  metabolism*
Saccharomyces cerevisiae / genetics,  metabolism*
Saccharomyces cerevisiae Proteins / genetics,  metabolism
Chemical
Reg. No./Substance:
0/PTH protein, human; 0/Parathyroid Hormone; 0/Recombinant Proteins; 0/Saccharomyces cerevisiae Proteins; EC 3.4.23.-/Aspartic Acid Endopeptidases; EC 3.4.23.-/MKC7 protein, S cerevisiae; EC 3.4.23.-/YPS3 protein, S cerevisiae; EC 3.4.23.25/YPS1 protein, S cerevisiae

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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