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Expression and Purification of Soluble Porcine Cystatin 11 in Pichia pastoris

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Abstract

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.

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Acknowledgments

This research was sponsored by the Key Scientific and Technological Project of Shanxi Province (Grant No. 20100312016). These experiments comply with the current laws of China.

Conflict of Interest

The authors have no conflict of interest to report.

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Authors and Affiliations

  1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi, 030801, People’s Republic of China

    Kuohai Fan, Junbing Jiang, Ruicheng Fan, Wei Yin, Yaogui Sun & Hongquan Li

  2. Transplantation Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA

    Zhirui Wang

  3. MGH-DF/HCC Recombinant Protein Expression and Purification Core, Boston, MA, USA

    Zhirui Wang

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  1. Kuohai Fan
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  2. Junbing Jiang
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  3. Zhirui Wang
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  4. Ruicheng Fan
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  5. Wei Yin
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  6. Yaogui Sun
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  7. Hongquan Li
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Correspondence to Hongquan Li.

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Fan, K., Jiang, J., Wang, Z. et al. Expression and Purification of Soluble Porcine Cystatin 11 in Pichia pastoris . Appl Biochem Biotechnol 174, 1959–1968 (2014). https://doi.org/10.1007/s12010-014-1148-z

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  • DOI: https://doi.org/10.1007/s12010-014-1148-z

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