Production and purification of recombinant human hepcidin-25 with authentic N and C-termini

Vignesh Narasimhan Janakiraman (1) , Charlotte Cabanne (1) , Wilfrid Dieryck (1) , Agnès Hocquellet (1) , Gilles Joucla (1) , Caroline Le Senechal (1) , Stéphane Chaignepain (2) , Patricia Costaglioli (1) , Xavier Santarelli (1) , Bertrand Garbay (1, 3, 4) , Abdelmajid Noubhani (1)

1 Biotechnologie des protéines recombinantes à visée santé
2 CBMN - Chimie et Biologie des Membranes et des Nanoobjets
3 LCPO - Laboratoire de Chimie des Polymères Organiques
4 Team 3 LCPO : Polymer Self-Assembly & Life Sciences

Hepcidin was first identified as an antimicrobial peptide present in human serum and urine. It was later demonstrated that hepcidin is the long-sought hormone that regulates iron homeostasis in mammals. Recombinant human Hepcidin-25 (Hepc25) was expressed in Pichia pastoris using a modified version of the pPICZ alpha A vector. Hepc25 was then purified by a simple two-step chromatographic process to obtain 1.9 mg of soluble recombinant human Hepc25 per liter of culture at 96% purity. The sequence of Hepc25 and the presence of four disulfide bridges were confirmed by mass spectrometry analyses, and the recombinant Hepc25 exhibited antibacterial activity. This protocol of production and purification is the first step toward the production of human Hepc25 at a greater scale. (C) 2015 Elsevier B.V. All rights reserved.

Mots clés: Hepcidin; P. pastoris; Recombinant expression; Expanded bed adsorption; IMAC

Domaines: Polymères

Vignesh Narasimhan Janakiraman, Charlotte Cabanne, Wilfrid Dieryck, Agnès Hocquellet, Gilles Joucla, et al.. Production and purification of recombinant human hepcidin-25 with authentic N and C-termini. Journal of Biotechnology, 2015, 195, pp.89-92. ⟨10.1016/j.jbiotec.2014.12.025⟩. ⟨hal-01373236⟩