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Reduction of aggregation of β-lactoglobulin during heating by dihydrolipoic acid

Published online by Cambridge University Press:  19 July 2013

Heni B Wijayanti ,
H Eustina Oh ,
Ranjan Sharma  and
Hilton C Deeth
Heni B Wijayanti
Affiliation:
School of Agriculture and Food Sciences, The University of Queensland, Brisbane QLD 4072, Australia
H Eustina Oh
Affiliation:
School of Agriculture and Food Sciences, The University of Queensland, Brisbane QLD 4072, Australia
Ranjan Sharma
Affiliation:
School of Agriculture and Food Sciences, The University of Queensland, Brisbane QLD 4072, Australia
Hilton C Deeth*
Affiliation:
School of Agriculture and Food Sciences, The University of Queensland, Brisbane QLD 4072, Australia
*
*For correspondence; e-mail: h.deeth@uq.edu.au
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Abstract

Prevention of the heat-induced aggregation of β-lactoglobulin (β-Lg) would improve the heat stability of whey proteins. The effects of lipoic acid (LA, or thioctic acid), in both its oxidised and reduced form (dihydrolipoic acid, DHLA), on heat-induced unfolding and aggregation of β-Lg were investigated. LA/DHLA was added to native β-Lg and the mixture was heated at 70, 75, 80 or 85 °C for up to 30 min at pH 6·8. The samples were analysed by Polyacrylamide Gel Electrophoresis (PAGE) and Size-exclusion HPLC (SE-HPLC). LA was not as effective as DHLA in reducing the formation of aggregates of heated β-Lg. Heating β-Lg with DHLA resulted in formation of more β-Lg monomers (due to dissociation of native dimers) and significantly less β-Lg aggregates, compared with heating β-Lg alone. The aggregates formed in the presence of DHLA were both covalently linked, via disulphide bonds, and non-covalently (hydrophobically) linked, but the amount of covalently linked aggregates was much less than when β-Lg was heated alone. The results suggest that DHLA was able to partially trap the reactive β-Lg monomer containing a free sulphydryl (−SH) group, by forming a ‘modified monomer’, and to prevent some sulphydryl−sulphydryl and sulphydryl−disulphide interactions that lead to the formation of covalently linked protein aggregates. The effects of DHLA were similar to those of N-ethylmaleimide (NEM) and dithio(bis)-p-nitrobenzoate (DTNB). However, the advantage of using DHLA over NEM and DTNB to lessen aggregation of β-Lg is that it is a food-grade compound which occurs naturally in milk.

Keywords

Aggregationβ-lactogloblinheatinglipoic aciddihydrolipoic acidmonomersdimersaggregatessulphydryl group
Type
Research Article
Information
Journal of Dairy Research , Volume 80 , Issue 4 , November 2013 , pp. 383 - 389
Copyright
Copyright © Proprietors of Journal of Dairy Research 2013 

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