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Biotechnology and Applied Biochemistry (2003) 37, (317–324) (Printed in Great Britain)

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Continuous production of a peptidic fraction containing the intermediate opioid peptide LVV-haemorphin-7 (LVVh-7) by peptic hydrolysis of bovine haemoglobin in a continuous membrane reactor

Romain Kapel, Renato Froidevaux, Naima Nedjar-Arroume, Anne Fertin-Bazus, Pascal Dhulster¹ and Didier Guillochon

Laboratoire de Technologie des Substances Naturelles, Bâtiment EUDIL/IAAL aile C, boulevard Paul Langevin, cité scientifique, 59655 Villeneuve d'Ascq, France

Key words: decolorization, haemorphins, ultrafiltration.

Abbreviations used: CR, regenerated cellulose; DH, degree of hydrolysis; IP, intermediate peptide; LVVh-7, LVV-haemorphin-7 (Leu-Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe); MALDI-MS, matrix-assisted laser-desorption–ionization MS; MMCO, molecular-mass cut-off; RP-, reversed-phase.

¹To whom correspondence should be addressed (e-mail pascal.dhulster@univ-lille1.fr).

Peptic hydrolysis of native bovine haemoglobin at pH 3 yields the LVV-haemorphin-7 (Leu-Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe; LVVh-7) opioid peptide corresponding to the residues-31–40 fragment of the b-chain of haemoglobin. This peptide is intermediate in the course of batch hydrolysis and is rapidly degraded. Indeed, it shows an optimum at 3% degree of hydrolysis (i.e. 2 min of reaction time). The hydrolysis was carried out in a continuous membrane reactor with a space time (ratio of the flux to the reactor volume) set to 2 min (corresponding to optimum LVVh-7 production). This process allows the continuous production of a constant fraction of intermediate peptides containing LVVh-7 for 48 min.

Received 9 September 2002/29 January 2003; accepted 31 January 2003

Published as Immediate Publication 31 January 2003, DOI 10.1042/BA20020078

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