An improved method for enhanced production and biological activity of human secretory leukocyte protease inhibitor (SLPI) in Pichia pastoris

作者：Li Zhiguo; Moy Allison; Gomez Seth R; Franz Andreas H; Lin Cereghino Joan; Lin Cereghino Geoff P^*

来源：Biochemical and Biophysical Research Communications, 2010, 402(3): 519-524.

DOI：10.1016/j.bbrc.2010.10.067

• Pichia pastoris
• Recombinant expression
• Protein disulfide isomerase
• Secretory leukocyte protease inhibitor
• Selectable markers

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 e

• 出版日期2010-11-19