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Open AccessArticle

High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli

1
Institute of Biotechnology and Department of Medical Science, National Tsing Hua University, Hsinchu 300, Taiwan
2
Department of Neurology, National Taiwan University Hospital, Hsinchu 300, Taiwan
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Molecules 2018, 23(4), 800; https://doi.org/10.3390/molecules23040800
Received: 24 January 2018 / Revised: 3 March 2018 / Accepted: 20 March 2018 / Published: 30 March 2018
P-113, which was originally derived from the human saliva protein histatin 5, is a histidine-rich antimicrobial peptide with the sequence AKRHHGYKRKFH. P-113 is currently undergoing phase II clinical trial as a pharmaceutical agent to fight against fungal infections in HIV patients with oral candidiasis. Previously, we developed a new procedure for the high-yield expression and purification of hG31P, an analogue and antagonist of human CXCL8. Moreover, we have successfully removed lipopolysaccharide (LPS, endotoxin) associated with hG31P in the expression with Escherichia coli. In this paper, we have used hG31P as a novel fusion protein for the expression and purification of P-113. The purity of the expressed P-113 is more than 95% and the yield is 4 mg P-113 per liter of E. coli cell culture in Luria-Bertani (LB) medium. The antimicrobial activity of the purified P-113 was tested. Furthermore, we used circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy to study the structural properties of P-113. Our results indicate that using hG31P as a fusion protein to obtain large quantities of P-113 is feasible and is easy to scale up for commercial production. An effective way of producing enough P-113 for future clinical studies is evident in this study. View Full-Text
Keywords: P-113; hG31P; expression; purification; NMR P-113; hG31P; expression; purification; NMR
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MDPI and ACS Style

Cheng, K.-T.; Wu, C.-L.; Yip, B.-S.; Yu, H.-Y.; Cheng, H.-T.; Chih, Y.-H.; Cheng, J.-W. High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli. Molecules 2018, 23, 800.

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