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In this study, we investigated structural aspects of covalent binding of food derived blue pigment phycocyanobilin (PCB) to bovine β-lactoglobulin (BLG), major whey protein, by spectroscopic, electrophoretic, mass spectrometry and computational methods. At physiological pH (7.2), we found that covalent pigment binding via free cysteine residue is slow ( k a  = 0.065 min −1 ), of moderate affinity ( K a  = 4 × 10 4  M −1 ), and stereo-selective. Binding also occurs at a broad pH range and under simulated gastrointestinal conditions. Adduct formation rises with pH, and in concentrated urea ( k a  = 0.101 min −1 ). The BLG-PCB adduct has slightly altered secondary and tertiary protein structure, and bound PCB has higher fluorescence and more stretched conformation than free chromophore. Combination of steered molecular dynamic for disulfide exchange, non-covalent and covalent docking, favours Cys119 residue in protein calyx as target for covalent BLG-PCB adduct formation. Our results suggest that this adduct can serve as delivery system of bioactive PCB. [ABSTRACT FROM AUTHOR]

The present study was aimed to evaluate the interaction between anthocyanins from black rice and soybean protein isolate (SPI) via non-covalent and covalent bindings and the impact of these interactions on the functional and conformational changes of soybean protein. The conformational changes of the protein structure with different concentrations of anthocyanins (0.05, 0.1%, and 0.2%) were analyzed using three-dimensional fluorescence and Fourier transform infrared spectroscopy. The anthocyanins were more likely to form covalent interactions with SPI instead of non-covalent interactions. The addition of anthocyanins changed the secondary structure of SPI with a decrease in β-sheets and an increase in β-turns and random coils. The emulsifying and foaming properties of SPI were improved after complexation with anthocyanins. This study might be useful for elucidating the mechanisms behind the binding of anthocyanins with SPI and the possible uses of SPI-anthocyanin complexes in food formulations. [ABSTRACT FROM AUTHOR]

Abstract: The effect of conjugating konjac glucomannan with fluorescein isothyocyanate (FITC) via covalent labelling on selected physicochemical properties of FITC and konjac was investigated using spectrophotometry, rheometry, and size exclusion chromatography (SEC). The binding of the lectin concanavalin A (ConA) to sugar residues of konjac was investigated using SEC. Covalent conjugation of konjac with FITC led to a shift in the absorbance spectrum peak of FITC to a lower wavelength and a decrease in the average molecular weight distribution of konjac. Furthermore, covalently labelled konjac showed reduced apparent viscosity compared to unlabelled konjac. ConA bound to the sugar residues in konjac. The potential of konjac–FITC covalent labelled conjugate or konjac–lectin labelled ConA complexes as fluorescent markers for localisation of konjac in a phase separated micellar casein–konjac mixture was investigated using confocal laser scanning microscopy. Results indicated that covalently labelled konjac has microscopic phase behaviour similar to that of un-labelled konjac and are therefore suitable for localising konjac glucomannan in a phase-separated micellar casein–konjac system. [Copyright &y& Elsevier]