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A simple method for the two‐step preparation of two pure haemorphins from a total haemoglobin peptic hydrolysate by conventional low‐pressure chromatographies
Author(s) -
Choisnard Luc,
Durand David,
VercaigneMarko Dominique,
NedjarArroume Naima,
Dhulster Pascal,
Guillochon Didier
Publication year - 2001
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20010039
Subject(s) - hydrolysate , chromatography , chemistry , elution , fractionation , sephadex , hydrolysis , enzyme , biochemistry
The development of a simple purification method for two haemorphins, VV‐haemorphin‐7 (Val‐Val‐Tyr‐Pro‐Trp‐Thr‐Gln‐Arg‐Phe) and VV‐haemorphin‐4 (Val‐Val‐Tyr‐Pro‐Trp‐Thr), from a total peptic haemoglobin hydrolysate is described. Cation‐exchange chromatography on CM‐Sephadex was used as a pre‐fractionation step for the hydrolysate. VV‐haemorphin‐7 and VV‐haemorphin‐4 were eluted in two different fractions. The second and final purification step was hydrophobic‐interaction chromatography on phenyl‐Sepharose, which allowed the isolation of the two pure haemorphins. Haemoglobin and globin hydrolysates were compared as starting materials. For easy recovery of haemorphins and easy adjustment of conditions for final purification, a volatile buffer, ammonium acetate buffer, pH 6.5, was employed. This process, which allowed the preparation of pure haemorphins from a total protein hydrolysate, could be scaled up and used in the food industry.