An improved method for enhanced production and biological activity of human secretory leukocyte protease inhibitor (SLPI) in Pichia pastoris
Document Type
Article
Publication Title
Biochemical and Biophysical Research Communications
Department
Chemistry
ISSN
0006-291X
Volume
402
Issue
3
DOI
10.1016/j.bbrc.2010.10.067
First Page
519
Last Page
524
Publication Date
11-19-2010
Abstract
The human secretory leukocyte protease inhibitor (SLPI) is an 11.7 kD cysteine-rich protein that has been shown to possess anti-protease, anti-inflammatory, and antimicrobial properties. By using a Pichia pastoris strain that overproduces protein disulfide isomerase (PDI), we obtained greater than fivefold higher levels of SLPI than in strains expressing normal levels of PDI and containing multiple copies of the SLPI gene. Elevated levels of PDI also enhanced the specific activity of the secreted SLPI by helping it achieve a proper tertiary structure. Mass spectrometry analysis indicated a greater number of disulfide bonds in the SLPI produced by the PDI overexpression strain compared to the SLPI produced in strains with normal PDI levels. Although others have utilized a similar strategy to increase yield, we believe that this is the first example of PDI overexpression being demonstrated to enhance the folding and thus increase the biological activity of a protein produced in the yeast P. pastoris.
Recommended Citation
Li, Z.,
Moy, A.,
Gomez, S. R.,
Franz, A. H.,
Lin-Cereghino, J.,
&
Lin-Cereghino, G. P.
(2010).
An improved method for enhanced production and biological activity of human secretory leukocyte protease inhibitor (SLPI) in Pichia pastoris.
Biochemical and Biophysical Research Communications, 402(3), 519–524.
DOI: 10.1016/j.bbrc.2010.10.067
https://scholarlycommons.pacific.edu/cop-facarticles/125
