NDLI: Effects of signal sequences and folding accessory proteins on extracellular expression of carboxypeptidase Y in recombinant Saccharomyces cerevisiae

Content Provider	Springer Nature Link
Author	Shin, So Yeon Bae, Yi Hyun Kim, Sun Ki Seong, Yeong Je Choi, Suk Hwan Kim, Kyoung Heon Park, Yong Cheol Seo, Jin Ho
Copyright Year	2013
Abstract	Carboxypeptidase Y (CPY) is a yeast vacuolar protease with useful applications including C-terminal sequencing of peptides and terminal modification of target proteins. To overexpress CPY with the pro-sequence (proCPY) encoded by the Saccharomyces cerevisiae PRC1 gene in recombinant S. cerevisiae, the proCPY gene was combined with the gene coding for a signal sequence of S. cerevisiae mating factor α (MFα), invertase (SUC2), or Kluyveromyces marxianus inulinase (INU1). Among the three constructs, the MFα signal sequence gave the best specific activity of extracellular CPY. To enhance the CPY expression level, folding accessory proteins of Kar2p, Pdi1p and Ero1p located in the S. cerevisiae endoplasmic reticulum were expressed individually and combinatorially. A single expression of Kar2p led to a 28 % enhancement in extracellular CPY activity, relative to the control strain of S. cerevisiae CEN.PK2-1D/p426Gal1-MFαCPY. Coexpression of Kar2p, Pdi1p and Ero1p gave a synergistic effect on CPY expression, of which activity was 1.7 times higher than that of the control strain. This work showed that engineering of signal sequences and protein-folding proteins would be helpful to overexpress yeast proteins of interest.
Starting Page	1065
Ending Page	1071
Page Count	7
File Format	PDF
ISSN	16157591
Journal	Bioprocess Engineering
Volume Number	37
Issue Number	6
e-ISSN	16157605
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2013-10-22
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Carboxypeptidase Y Saccharomyces cerevisiae Signal sequence Folding accessory protein Fermentation Biotechnology Industrial and Production Engineering Environmental Engineering/Biotechnology Industrial Chemistry/Chemical Engineering Food Science
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Biotechnology

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