https://scholars.lib.ntu.edu.tw/handle/123456789/432012
Title: | High level expression and purification of the clinically active antimicrobial peptide P-113 in Escherichia coli | Authors: | B.-S. Yip Cheng, Kuang-Ting ; Wu, Chih-Lung ; Yu, Hui-Yuan ; Cheng, Hsi-Tsung ; Chih, Ya-Han ; Cheng, Jya-Wei |
Keywords: | Expression; HG31P; NMR; P-113; Purification | Issue Date: | 2018 | Journal Volume: | 23 | Journal Issue: | 4 | Start page/Pages: | 800 | Source: | Molecules | Abstract: | 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. ? 2018 by the authors. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85044830107&doi=10.3390%2fmolecules23040800&partnerID=40&md5=4631ee298235dedf22687fb5190c5666 https://scholars.lib.ntu.edu.tw/handle/123456789/432012 |
DOI: | 10.3390/molecules23040800 | SDG/Keyword: | antimicrobial cationic peptide; fusion protein; histatin; HTN3 protein, human; biosynthesis; chemistry; Escherichia coli; gene expression; genetics; growth, development and aging; human; isolation and purification; Antimicrobial Cationic Peptides; Escherichia coli; Gene Expression; Histatins; Humans; Recombinant Fusion Proteins |
Appears in Collections: | 醫學院附設醫院 (臺大醫院) |
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