Expression and characterization of recombinant human secretory leukocyte protease inhibitor (SLPI) protein from Pichia pastoris

Authors

Zhiguo Li, University of the Pacific
Allison Moy, University of the Pacific
Kirti Sohal, University of the Pacific
Carolyn Dam, University of the Pacific
Peter Kuo, University of the Pacific
James Whittaker, Oregon Health and Science University
Mei Whittaker, Oregon Health and Science University
Nejat Düzgünes, University of the PacificFollow
Krystyna Konopka, University of the Pacific
Andreas H. Franz, University of the PacificFollow
Joan Lin-Cereghino, University of the PacificFollow
Geoff P. Lin-Cereghino, University of the PacificFollow

Document Type

Article

Publication Title

Protein Expression and Purification

Department

Chemistry

ISSN

1046-5928

Volume

67

Issue

2

DOI

10.1016/j.pep.2009.06.001

First Page

175

Last Page

181

Publication Date

10-1-2009

Abstract

The human secretory leukocyte protease inhibitor (SLPI) has been shown to possess anti-protease, anti-inflammatory and antimicrobial properties. Its presence in saliva is believed to be a major deterrent to oral transmission of human immunodeficiency virus-1. The 11.7 kDa peptide is a secreted, nonglycosylated protein rich in disulfide bonds. Currently, recombinant SLPI is only available as an expensive bacterial expression product. We have investigated the utility of the methylotrophic yeast Pichia pastoris to produce and secrete SLPI with C-terminal c-myc and polyhistidine tags. The post-transformational vector amplification protocol was used to isolate strains with increased copy number, and culturing parameters were varied to optimize SLPI expression. Modification of the purification procedure allowed the secreted, recombinant protein to be isolated from the cell-free fermentation medium with cobalt affinity chromatography. This yeast-derived SLPI was shown to have an anti-protease activity comparable to the commercially available bacterial product. Thus, P. pastoris provides an efficient, cost-effective system for producing SLPI for structure function analysis studies as well as a wide array of potential therapeutic applications.

Recommended Citation

Li, Z., Moy, A., Sohal, K., Dam, C., Kuo, P., Whittaker, J., Whittaker, M., Düzgünes, N., Konopka, K., Franz, A. H., Lin-Cereghino, J., & Lin-Cereghino, G. P. (2009). Expression and characterization of recombinant human secretory leukocyte protease inhibitor (SLPI) protein from Pichia pastoris. Protein Expression and Purification, 67(2), 175–181. DOI: 10.1016/j.pep.2009.06.001
https://scholarlycommons.pacific.edu/cop-facarticles/112