Chemical and Biomolecular Engineering - Research Publications

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    Effect of rennet on the composition, proteolysis and microstructure of reduced-fat Cheddar cheese during ripening
    Soodam, K ; Ong, L ; Powell, IB ; Kentish, SE ; Gras, SL (SPRINGER FRANCE, 2015-09-01)
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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 addition of calcium chloride in combination with a lower draining pH to change the microstructure and improve fat retention in Cheddar cheese
    Ong, L ; Soodam, K ; Kentish, SE ; Powell, IB ; Gras, SL (Elsevier, 2015-07-01)
    Calcium chloride addition and the whey draining pH are known to impact on cheese making. The effect of 100 or 300 mg kg−1 calcium chloride (CaCl2) and the whey draining pH (6.2 or 6.0) on the microstructure of Cheddar cheese was assessed using confocal and cryo scanning electron microscopy. The gel made with 300 mg kg−1 CaCl2 was found to have a denser protein network and smaller pores than the gel with lower or no CaCl2 addition. CaCl2 addition reduced fat lost to the sweet whey. The texture of the cheeses with a lower draining pH was harder and moisture content lower. Our results show that the combination of calcium addition and lower draining pH could be used to increase network formation at the early stages of cheese making to improve fat retention while maintaining a similar level of total calcium in the final cheese.
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    Small scale production of cream cheese: A comparison of batch centrifugation and cloth bag methods
    Ong, L ; Kentish, SE ; Gras, SL (Elsevier, 2018-06-01)
    Cream cheese production is well established at large scale but an effective small scale process could facilitate higher throughput and lower the cost of experimental studies. Whey was separated using centrifugation or the cloth bag method and the effect of heating prior to separation examined. Heat treatment and centrifugation resulted in cream cheese with a microstructure, composition and rheological properties comparable with that of a commercial scale cream cheese. Heating was necessary to achieve effective separation, the desired product microstructure and an adequate firmness and viscosity, with the heat induced denaturation of some whey proteins contributing to these properties. Whilst both whey separation methods resulted in a similar microstructure, centrifugation led to less fat loss and an optimal product. These data provide new insights into the development of cream cheese microstructure and provide a route to further understand and optimise this process.
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    The Microstructure and Physicochemical Properties of Probiotic Buffalo Yoghurt During Fermentation and Storage: a Comparison with Bovine Yoghurt
    Hanh, THN ; Ong, L ; Lefevre, C ; Kentish, SE ; Gras, SL (Springer, 2014-04-01)
    The physicochemical and rheological properties of yoghurt made from unstandardised unhomogenised buffalo milk were investigated during fermentation and 28 days of storage and compared to the properties of yoghurt made from homogenised fortified bovine milk. A number of differences observed in the gel network can be linked to differences in milk composition. The microstructure of buffalo yoghurt, as assessed by confocal laser scanning microscopy (CLSM) and cryo scanning electron microscopy (cryo-SEM), was interrupted by large fat globules and featured more serum pores. These fat globules have a lower surface area and bind less protein than the homogenised fat globules in bovine milk. These microstructural differences likely lead to the higher syneresis observed for buffalo yoghurt with an increase from 17.4 % (w/w) to 19.7 % (w/w) in the weight of whey generated at days 1 and 28 of the storage. The higher concentration of total calcium in buffalo milk resulted in the release of more ionic calcium during fermentation. Gelation was also slower but the strength of the two gels was similar due to similar protein and total solids concentrations. Buffalo yoghurt was more viscous, less able to recover from deformation and less Newtonian than bovine yoghurt with a thixotropy of 3,035 Pa.s−1 measured for buffalo yoghurt at the end of the storage, at least four times higher than the thixotropy of bovine yoghurt. While the titratable acidity, lactose consumption and changes in organic acid concentrations were similar, differences were recorded in the viability of probiotic bacteria with a lower viability of Lactobacillus acidophilus of 5.17 log (CFU/g) recorded for buffalo yoghurt at day 28 of the storage. Our results show that factors other than the total solids content and protein concentration of milk affect the structural properties of yoghurt. They also illustrate the physicochemical reasons why buffalo and bovine yoghurt are reported to have different sensory properties and provide insight into how compositional changes can be used to alter the microstructure and properties of dairy 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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    The dynamics of the biological membrane surrounding the buffalo milk fat globule investigated as a function of temperature
    Nguyen, HTH ; Madec, M-N ; Ong, L ; Kentish, SE ; Gras, SL ; Lopez, C (Elsevier, 2016-08-01)
    The biological membrane surrounding fat globules in milk (the MFGM) is poorly understood, despite its importance in digestion and in determining the properties of fat globules. In this study, in situ structural investigations of buffalo MFGM were performed as a function of temperature (4–60 °C), using confocal microscopy. We demonstrate that temperature and rate of temperature change affected the lipid domains formed in the MFGM with the lateral segregation (i) of high Tm lipids and cholesterol in a Lo phase for both T < Tm and T > Tm and (ii) of high Tm lipids in a gel phase for T < Tm. Rapid cooling favours nucleation, while slow cooling favours growth, leading to the formation of small and large lipid domains, respectively. Changes in the interfacial properties of the MFGM, as a function of temperature, could modulate the functions of fat globules during processing and digestion.
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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.