Expression and Purification of Soluble Porcine Cystatin 11 in Pichia pastoris

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• 作者：Kuohai Fan (1)
  Junbing Jiang (1)
  Zhirui Wang (2) (3)
  Ruicheng Fan (1)
  Wei Yin (1)
  Yaogui Sun (1)
  Hongquan Li (1)
  
• 关键词：Antimicrobial activity ; Pichia pastoris ; Expression ; Purification ; Porcine CST11
• 刊名：Applied Biochemistry and Biotechnology
• 出版年：2014
• 出版时间：November 2014
• 年：2014
• 卷：174
• 期：5
• 页码：1959-1968
• 全文大小：783 KB
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• 作者单位：Kuohai Fan (1)
  Junbing Jiang (1)
  Zhirui Wang (2) (3)
  Ruicheng Fan (1)
  Wei Yin (1)
  Yaogui Sun (1)
  Hongquan Li (1)
  
  1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi, 030801, People鈥檚 Republic of China
  2. Transplantation Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
  3. MGH-DF/HCC Recombinant Protein Expression and Purification Core, Boston, MA, USA
  
• ISSN：1559-0291

文摘

Cystatin 11 (CST11) belongs to the cystatin type 2 family of cysteine protease inhibitors and exhibits antimicrobial activity in vitro. In this study, we describe the expression and purification of recombinant porcine CST11 in the Pichia pastoris system. We then assess its antimicrobial activity against Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus subtilis by liquid growth inhibition assay. Kinetic studies indicate that the recombinant porcine CST11 has high potency against E. coli and S. aureus. Scanning electronic microscope analysis showed that CST11 might be targeting the bacterial membrane and, thus, could potentially be developed as a therapeutic agent for inhibiting microbe infection without the risk of antibiotic resistance.