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Chymotrypsin selectively digests β-lactoglobulin in whey protein isolate away from enzyme optimal conditions: Potential for native α-lactalbumin purification

Published online by Cambridge University Press:  21 September 2012

Katarina Lisak ,
Jose Toro-Sierra ,
Ulrich Kulozik ,
Rajka Božanić  and
Seronei Chelulei Cheison
Katarina Lisak*
Affiliation:
Laboratory for Milk Technology and Dairy Products, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, HR-10000, Zagreb, Croatia
Jose Toro-Sierra
Affiliation:
Chair for Food Process Engineering and Dairy Technology Department, ZIEL Technology Section, Technische Universität München, Weihenstephaner Berg 1, D-85354 Freising, Germany
Ulrich Kulozik
Affiliation:
Chair for Food Process Engineering and Dairy Technology Department, ZIEL Technology Section, Technische Universität München, Weihenstephaner Berg 1, D-85354 Freising, Germany
Rajka Božanić
Affiliation:
Laboratory for Milk Technology and Dairy Products, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, HR-10000, Zagreb, Croatia
Seronei Chelulei Cheison
Affiliation:
Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL)-Junior Research Group: Bioactive Peptides and Protein Technology, Technische Universität München, Weihenstephaner Berg 1, D-85354 Freising, Germany School of Public Health and Community Development, Maseno University, Private Bag, Kisumu, Kenya
*
*For correspondence; e-mail: [email protected]
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Abstract

The present study examines the resistance of the α-lactalbumin to α-chymotrypsin (EC 3.4.21.1) digestion under various experimental conditions. Whey protein isolate (WPI) was hydrolysed using randomised hydrolysis conditions (5 and 10% of WPI; pH 7·0, 7·8 and 8·5; temperature 25, 37 and 50 °C; enzyme-to-substrate ratio, E/S, of 0·1%, 0·5 and 1%). Reversed-phase high performance liquid chromatography (RP-HPLC) was used to analyse residual proteins. Heat, pH adjustment and two inhibitors (Bowman–Birk inhibitor and trypsin inhibitor from chicken egg white) were used to stop the enzyme reaction. While operating outside of the enzyme optimum it was observed that at pH 8·5 selective hydrolysis of β-lactoglobulin was improved because of a dimer-to-monomer transition while α-la remained relatively resistant. The best conditions for the recovery of native and pure α-la were at 25 °C, pH 8·5, 1% E/S ratio, 5% WPI (w/v) while the enzyme was inhibited using Bowman–Birk inhibitor with around 81% of original α-la in WPI was recovered with no more β-lg. Operating conditions for hydrolysis away from the chymotrypsin optimum conditions offers a great potential for selective WPI hydrolysis, and removal, of β-lg with production of whey protein concentrates containing low or no β-lg and pure native α-la. This method also offers the possibility for production of β-lg-depleted milk products for sensitive populations.

Keywords

α-chymotrypsinselective whey protein hydrolysisβ-lactoglobulinα-lactalbumin recoveryBowman–Birk inhibitorchicken egg-white inhibitor
Type
Research Article
Information
Journal of Dairy Research , Volume 80 , Issue 1 , February 2013 , pp. 14 - 20
Copyright
Copyright © Proprietors of Journal of Dairy Research 2012

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