Structural characterization of bovine β-lactoglobulin-galactose/ tagatose maillard complexes by electrophoretic, chromatographic, and spectroscopic methods

To investigate the influence of the type of carbonyl group of the sugar on the structural changes of proteins during glycation, an exhaustive structural characterization of glycated β-lactoglobulin with galactose (aldose) and tagatose (ketose) has been carried out. Conjugates were prepared via Maillard reaction at 40 and 50°C, pH 7, and aw = 0.44. The progress of the Maillard reaction was followed by indirect formation of Amadori and Heyns compounds, advanced glycation end products, and brown polymers. The structural characterization of glycoconjugates was conducted by using a number of analytical techniques such as RP-HPLC, isoelectric focusing, MALDI-ToF, SDS-PAGE, size exclusion chromatography, and spectrofluorimetry (tryptophan fluorescence). In addition, the surface hydrophobicity of the β-lactoglobulin glycoconjugates was also assessed. The results showed a higher reactivity of galactose than tagatose to form the glycoconjugates, probably due to the higher electrophilicity of the aldehyde group. At 40°C, more aggregation was produced when β-lactoglobulin was conjugated with tagatose as compared to galactose. However, at 50°C hardly any difference was observed in the aggregation produced by galactose and tagatose. These results afford more insight into the importance of the functional group of the carbohydrate moiety during the formation of protein-carbohydrate conjugates via Maillard reaction.This work was supported by Projects AGL-2004-07227-C02-02 and Consolider Ingenio 2010 FUNC-FOOD CSD2007-00063 (Ministerio de Educación y Ciencia) and ALIBIRD S-0505/AGR/000153 (Comunidad de Madrid). M.C.-M. thanks the CSIC for an I3P Ph.D. grant.Peer Reviewe