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Expression and characterization of recombinant human secretory leukocyte protease inhibitor (SLPI) protein from Pichia pastoris
Protein Expression & Purification
  • 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 Pacific
  • Krystyna Konopka, University of the Pacific
  • Andreas H. Franz, University of the Pacific
  • Joan Lin-Cereghino, University of the Pacific
  • Geoff P. Lin-Cereghino, University of the Pacific
Document Type
Article
Department
Chemistry
DOI
10.1016/j.pep.2009.06.001
Publication Date
10-1-2009
Disciplines
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.
Citation Information
Zhiguo Li, Allison Moy, Kirti Sohal, Carolyn Dam, et al.. "Expression and characterization of recombinant human secretory leukocyte protease inhibitor (SLPI) protein from Pichia pastoris" Protein Expression & Purification Vol. 67 Iss. 2 (2009) p. 175 - 181 ISSN: 1046-5928
Available at: http://works.bepress.com/andreas-franz/123/