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Proteolytic specificity of chymosin on bovine αs1,-casein

Published online by Cambridge University Press:  01 June 2009

Paul L. H. McSweeney ,
Norman F. Olson ,
Patrick F. Fox ,
Aine Healy  and
Peter Højrup
Paul L. H. McSweeney
Affiliation:
Center for Dairy Research, University of Wisconsin-Madison, Madison, WI 53706, USA
Norman F. Olson
Affiliation:
Center for Dairy Research, University of Wisconsin-Madison, Madison, WI 53706, USA
Patrick F. Fox
Affiliation:
Department of Food Chemistry, University College, Cork, Irish Republic National Food Biotechnology Centre, University College, Cork, Irish Republic
Aine Healy
Affiliation:
National Food Biotechnology Centre, University College, Cork, Irish Republic
Peter Højrup
Affiliation:
Department of Molecular Biology, University of Odense, Campusvej 55, DK-5230, Odense M., Denmark
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Summary

The proteolytic specificity of chymosin (EC 3.4.23.4) on bovine αs1-casein at 30°C in phosphate buffer, pH 6·5 and at pH 5·2 in the presence of 5% (w/v) NaCl was investigated. Peptides (pH 4·6-soluble) were isolated by reversed-phase HPLC and identified from their amino acid sequence; the identity of some peptides was confirmed by mass spectrometry and/or amino acid composition. The small peptides produced at pH 6·5 were Arg1–Phe23, Phe24–Phe28, Phe24–Leu40(?), Phe150–Phe153, Phe150–Leu156, Tyr154–Tyr159, Tyr154–Trp164, Asp157–Trp164 and Tyr165–Trp199. The same peptides, except Tyr154–Trp164, were produced at pH 5·2 in the presence of NaCl and, in addition, the peptides Arg1–Leu11, Phe24–Phe32, Lys102–Leu142, Ala143-Leu149 and Tyr165-Phe179. The rates of production of individual peptides differed under the two conditions studied but Arg1-Phe23 and Tyr165–Trp199 were the first and second peptides produced under both conditions. Pathways are proposed to interpret the proteolysis of αs1-casein in solution under the conditions of this study.

Type
Original Articles
Information
Journal of Dairy Research , Volume 60 , Issue 3 , August 1993 , pp. 401 - 412
Copyright
Copyright © Proprietors of Journal of Dairy Research 1993

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