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The aim of this work was the expression, as secreted peptide, purification, and characterization of a new recombinant polyHis\u2010tagged camel hepcidin (HepcD\u2010His) in yeast <jats:italic>Pichia pastoris<\/jats:italic>. The use of this eukaryotic expression system, for the production of HepcD\u2010His, having 6 histidine residues at its C terminus, was simpler and more efficient compared with the use of the prokaryotic system <jats:italic>Escherichia<\/jats:italic>\u00a0<jats:styled-content style=\"fixed-case\"><jats:italic>coli<\/jats:italic><\/jats:styled-content>. Indeed, a single purification step was required to isolate the soluble hepcidin with purity estimated more that 94% and a yield of 2.8 against 0.2\u00a0mg\/L for the <jats:styled-content style=\"fixed-case\"><jats:italic>E\u00a0coli<\/jats:italic><\/jats:styled-content> system. Matrix\u2010assisted laser desorption\/ionization time\u2010of\u2010flight (TOF)\/TOF mass spectrometry of the purified HepcD\u2010His showed 2 major peaks at <jats:italic>m<\/jats:italic>\/<jats:italic>z<\/jats:italic> 4524.64 and 4634.56 corresponding to camel hepcidin with 39 and 40 amino acids. Evaluation of disulfide bond connectivity with the Ellman method showed an absence of free thiol groups, testifying that the 8 cysteine residues in the peptide are displayed, forming 4 disulfide bridges. Circular dichroism spectroscopy showed that camel hepcidin structure was significantly modified at high temperature of 90\u00b0C and returns to its original structure when incubation temperature drops back to 20\u00b0C. Interestingly, this peptide showed also a greater bactericidal activity, at low concentration of 9.5\u03bcM, against <jats:italic>E\u00a0coli<\/jats:italic>, than the synthetic analog DH3. Thus, the production, at a large scale, of the recombinant camel hepcidin, HepcD\u2010His, may be helpful for future therapeutic applications including bacterial infection diseases.<\/jats:p>","DOI":"10.1002\/jmr.2561","type":"journal-article","created":{"date-parts":[[2016,8,10]],"date-time":"2016-08-10T09:33:01Z","timestamp":1470821581000},"source":"Crossref","is-referenced-by-count":6,"title":["Recombinant overexpression of camel hepcidin cDNA in <i>Pichia pastoris<\/i>: purification and characterization of the polyHis\u2010tagged peptide HepcD\u2010His"],"prefix":"10.1002","volume":"30","author":[{"given":"Mohamed","family":"Boumaiza","sequence":"first","affiliation":[{"name":"Laboratoire d'ing\u00e9nierie des prot\u00e9ines et des mol\u00e9cules bioactives (LIP\u2010MB), Institut National des Sciences Appliqu\u00e9es et de Technologie Universit\u00e9 de Carthage  BP 676 1080 Tunis Cedex Tunis 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Pharmacologiques et Toxicologiques, UMR 8601 Universit\u00e9 Paris Descartes, CNRS  45 rue des Saints P\u00e8res 75270 Paris Cedex 06 Paris France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Gianoncelli","sequence":"additional","affiliation":[{"name":"Department of Molecular and Translational Medicine, DMMT University of Brescia  Viale Europa 11 25123 Brescia Brescia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanna","family":"Longhi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Translational Medicine, DMMT University of Brescia  Viale Europa 11 25123 Brescia Brescia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando","family":"Carmona","sequence":"additional","affiliation":[{"name":"Department of Molecular and Translational Medicine, DMMT University of Brescia  Viale Europa 11 25123 Brescia Brescia 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