title: Reduction of aggregation of β-lactoglobulin during heating by dihydrolipoic acid.
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.
authors.affiliation: School of Agriculture and Food Sciences, The University of Queensland, Brisbane QLD 4072, Australia.
authors.forename: Heni B
authors.initials: HB
authors.lastname: Wijayanti
authors.forename: H Eustina
authors.initials: HE
authors.lastname: Oh
authors.forename: Ranjan
authors.initials: R
authors.lastname: Sharma
authors.forename: Hilton C
authors.initials: HC
authors.lastname: Deeth
mesh_terms: D000818:Animals; D002417:Cattle; D004228:Dithionitrobenzoic Acid / Q000737:chemistry; D005033:Ethylmaleimide / Q000737:chemistry; D006361:Heating; D007782:Lactoglobulins / Q000737:chemistry*; D008892:Milk / Q000737:chemistry; D008894:Milk Proteins; D011489:Protein Denaturation; D008063:Thioctic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D000067816:Whey Proteins
chemical_list: D007782:Lactoglobulins; D008894:Milk Proteins; D000067816:Whey Proteins; D008063:Thioctic Acid; C015144:dihydrolipoic acid; D004228:Dithionitrobenzoic Acid; D005033:Ethylmaleimide