Chemical and Biomolecular Engineering - Research Publications

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    A proteomic characterization shows differences in the milk fat globule membrane of buffalo and bovine milk
    Nguyen, HTH ; Ong, L ; Hoque, A ; Kentish, SE ; Williamson, N ; Ang, C-S ; Gras, SL (Elsevier, 2017-09-01)
    The proteins of the milk fat globule membrane (MFGM) have a number of functions, such as the regulation of milk fat secretion and metabolism, the uptake and transportation of fatty acids in the intestine, and potential protection from bacterial or viral infection. While the proteome of the MFGM in bovine milk has been extensively characterized, knowledge of these proteins in buffalo milk is limited. In this study, a proteomic approach was used to characterize the proteome of the buffalo MFGM. Multiple extraction techniques were used to increase the number of proteins identified, while label free relative quantitative liquid chromatography tandem mass spectrometry was used for comparison between the buffalo and bovine MFGM proteomes. A total of 220 buffalo MFGM proteins and 234 bovine MFGM proteins were identified after being filtered from the initial dataset of 757 and 680 proteins, respectively. A sixfold higher concentration of xanthine oxidoreductase was identified per mass of buffalo MFGM protein extracted, together with significantly greater concentrations of platelet glycoprotein 4, heat shock cognate and calcineurin B homologous protein. The expression of xanthine oxidoreductase in the MFGM of buffalo milk, which can affect milk shelf-life and flavor, was confirmed by Western blot analysis and a heterogeneous distribution of this protein observed in situ on the surface of the MFGM. The high concentration of fat in buffalo milk, together with the differences in the MFGM proteome provide insights into the differences in nutritional profile, biological function and properties of these two milk products.
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    The Effect of Fermentation Temperature on the Microstructure, Physicochemical and Rheological Properties of Probiotic Buffalo Yoghurt
    Nguyen, HTH ; Ong, L ; Kentish, SE ; Gras, SL (Springer, 2014-09)
    The properties of buffalo and bovine milk differ and the procedures developed to make bovine yoghurt may require optimisation for the production of buffalo yoghurt. This study aimed to apply cryo-scanning electron microscopy and confocal laser scanning microscopy to determine the optimal temperature for processing buffalo yoghurt. Milk was fermented at three different temperatures (37, 40 and 43 °C), stored for 28 days and the yoghurt microstructure, physicochemical and rheological properties assessed. Yoghurt fermented at 37 °C had a compact microstructure and the probiotic Lactobacillus acidophilus La-5 was more viable on storage. In contrast, yoghurt produced from a faster fermentation at 43 °C was firmer with a more porous microstructure that exhibited a higher degree of syneresis. The rheological properties during storage including the thixotropy, consistency coefficient and flow behaviour index were not significantly affected by temperature nor were the concentration of lactose, ionic calcium or titratable acidity. This study shows how changes to processing can be used to alter the microstructure of buffalo products and suggests that a decrease in fermentation temperature could be used to improve the quality of buffalo yoghurt.
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    Microstructure and physicochemical properties reveal differences between high moisture buffalo and bovine Mozzarella cheeses
    Nguyen, HTH ; Ong, L ; Lopez, C ; Kentish, SE ; Gras, SL (Elsevier, 2017-12-01)
    Mozzarella cheese is a classical dairy product but most research to date has focused on low moisture products. In this study, the microstructure and physicochemical properties of both laboratory and commercially produced high moisture buffalo Mozzarella cheeses were investigated and compared to high moisture bovine products. Buffalo and bovine Mozzarella cheeses were found to significantly differ in their microstructure, chemical composition, organic acid and proteolytic profiles but had similar hardness and meltability. The buffalo cheeses exhibited a significantly higher ratio of fat to protein and a microstructure containing larger fat patches and a less dense protein network. Liquid chromatography mass spectrometry detected the presence of only β-casein variant A2 and a single β-lactoglobulin variant in buffalo products compared to the presence of both β-casein variants A1 and A2 and β-lactoglobulin variants A and B in bovine cheese. These differences arise from the different milk composition and processing conditions. The differences in microstructure and physicochemical properties observed here offer a new approach to identify the sources of milk used in commercial cheese products.
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    Buffalo milk fat globules and their biological membrane: in situ structural investigations
    Nguyen, HTH ; Ong, L ; Beaucher, E ; Madec, M-N ; Kentish, SE ; Gras, SL ; Lopez, C (Elsevier, 2015-01-01)
    Milk fat globules and their surrounding biological membrane (the MFGM) are not well understood despite the importance of these milk components in human nutrition and the role of fat globules in determining the properties of dairy products. The objectives of this study were to investigate these unique colloidal assemblies and the microstructure of the MFGM in buffalo milk, which is the second largest global source of dairy products. In-situ structural investigations were performed at room temperature using confocal microscopy with multiple fluorescent probes (Nile Red, Rh-DOPE, the lectin WGA-488). Microscopic observations showed cytoplasmic crescents around fat globules and the heterogeneous distribution of glycosylated molecules and polar lipids with the occurrence of lipid domains. The lipid domains in the buffalo MFGM appear to form by the segregation of lipids with a high phase transition temperature (e.g. sphingomyelin and saturated phosphatidylcholine molecular species) and cholesterol resulting in a gel phase or a Lo phase forming circular domains. The structure of the buffalo MFGM results from a non-random mixing of components, consistent with observations for other species. Structural heterogeneities of the MFGM could affect the processability of buffalo fat globules and the bioavailability of milk lipids.
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    Homogenisation improves the microstructure, syneresis and rheological properties of buffalo yoghurt
    Nguyen, HTH ; Ong, L ; Kentish, SE ; Gras, SL (ELSEVIER SCI LTD, 2015-07)
    Homogenisation is known to improve the textural properties of bovine yoghurt but the potential of this processing step has not been systematically explored for buffalo yoghurt. In this study, buffalo milk was homogenised at 80 bar or 160 bar and the effect on the properties of buffalo yoghurt examined. The microstructure of both buffalo yoghurt samples produced from homogenised milk was significantly altered, forming a more interconnected protein network with smaller embedded fat globules. These structural changes resulted in a significant decrease in syneresis and hysteresis area and led to a considerable increase in the storage modulus, gel firmness and flow behaviour index. A higher homogenisation pressure of 160 bar resulted in a lower gel firmness and storage modulus, possibly due to the presence of bigger fat–protein clusters within the homogenised milk. A homogenisation pressure of 80 bar could be optimal for improving the quality of buffalo yoghurt.