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Information regarding protein expression and phosphorylation modifications in the bovine milk fat globule membrane is scarce, particularly throughout various lactation periods. This study employed a complete proteome and phosphoproteome between bovine colostrum and mature milk. A total of 11 proteins were seen in both protein expression and phosphorylation levels. There were 400 proteins identified in only protein expression, and 104 phosphoproteins identified in only phosphorylation levels. A total of 232 significant protein characteristics were identified within the proteome and significant phosphorylation sites within 86 phosphoproteins of the phosphoproteome. Biological activities and pathways primarily exhibited associations with the immune system. Simultaneously, a comprehensive analysis of proteins and phosphorylation sites using a multi-omics approach. Hence, the data we have obtained has the potential to expand our understanding of how the bovine milk fat globule membrane might be utilized as a beneficial component in dairy products. • A total of 411 MFGM proteins and 197 phosphorylation sites on 115 phosphoproteins were identified from bovine milk. • There were 11 shared MFGM proteins between proteome and phosphoproteome. • 232 MFGM proteins and 122 phosphorylation sites on 86 phosphoproteins were significantly different features. • Proteins and phosphoproteins were mainly involved in immune-related functions and pathways. • Comprehensive characterization and relationships of proteins and phosphorylation sites revealed by multi-omics analysis. [ABSTRACT FROM AUTHOR]

The human milk fat globule membrane (hMFGM) and Lactobacillus modulate the infant's gut and benefit health. Hence, the current study assesses the probiotic potential of Lactiplantibacillus plantarum (MRK3) , Limosilactobacillus ferementum (MK1) isolated from infant feces, and its interaction with hMFGM during conditions mimicking infant digestive tract. Both strains showed high tolerance to gastrointestinal conditions, cell surface hydrophobicity, and strong anti-pathogen activity against Staphylococcus aureus. During digestion, hMFGM significantly exhibited xanthine oxidase activity, membrane roughness, and surface topography. In the presence of hMFGM, survival of MRK3 was higher than MK1, and electron microscopic observation revealed successful entrapment of MRK3 in the membrane matrix throughout digestion. Interestingly, probiotic-membrane matrix interaction showed significant synergy to alleviate oxidative stress and damage induced by cell-free supernatant of Escherichia coli in Caco-2 cells. Our results show that a probiotic-encapsulated membrane matrix potentially opens the functional infant formula development pathway. [Display omitted] • L. plantarum and L. fermentum exhibit high tolerance against gastrointestinal stress. • L. plantarum incorporated fat globules increase protection during gastrointestinal stress. • hMFGM enhances the survival of probiotics in a strain-dependent manner during in vitro digestion. • Probiotic-membrane interaction improves oxidation and neonatal gut-derived infection. [ABSTRACT FROM AUTHOR]

• MFGM changed the rheological properties of the emulsion. • MFGM promoted the stabilization of fat globules in the emulsion by its stronger emulsification ability. • MFGM improves the physicochemical properties and surface composition of the powder. • It is feasible to improve powder properties by enhancing emulsion stability. The milk fat globules in infant formula (IF) are encapsulated by a component known as milk fat globule membrane (MFGM). However, it is currently unclear whether the improved emulsion stability of MFGM can have a profound effect on the finished IF. Therefore, this study investigated the effects of MFGM on the particle size, stability, rheology, and microstructure of emulsions prepared by dairy ingredients via wet mixing. Further, IF were processed using such emulsions, the physicochemical properties, surface composition of the powders were examined. The results showed that MFGM reduced the particle size of the emulsion, increased the viscosity, and improved the microstructure of the MFGM. Furthermore, MFGM reduced the moisture content of the powder, increased the glass transition temperature, and reduced the presence of surface fat. In conclusion, the addition of MFGM enhance the finished powder stability by improving the emulsion stability prepared during IF manufacturing. [ABSTRACT FROM AUTHOR]

• 3917 and 3966 MFGM proteins were identified in PC and PM milk. • 3807 common MFGM proteins were found in both groups. • 303 significant differentially expressed MFGM proteins were identified. Milk fat globule membrane (MFGM) proteins are nutritional components with various biological functions. This study aimed to analyze and compare MFGM proteins in porcine colostrum (PC) and porcine mature milk (PM), via label-free quantitative proteomics. In total, 3917 and 3966 MFGM proteins were identified in PC and PM milk, respectively. A total of 3807 common MFGM proteins were found in both groups, including 303 significant differentially expressed MFGM proteins. Gene Ontology (GO) analysis revealed that the differentially expressed MFGM proteins were mainly related to the cellular process, cell, and binding. The dominant pathway of the differentially expressed MFGM proteins was related to the phagosome according to Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. These results reveal crucial insights into the functional diversity of MFGM proteins in porcine milk during lactation and provide theoretical guidance for the development of MFGM proteins in the future. [ABSTRACT FROM AUTHOR]

[Display omitted] • Human MFGM lipids were compared with four dietary lipid sources via lipidomics. • A total of 45 lipid classes and 5048 lipid species were identified in all samples. • Glycerophospholipid metabolism was a key lipid metabolic pathway. • The study results can provide a theoretical basis for developing infant formula. Milk fat globule membrane (MFGM) contains lipids, which are essential for promoting infant brain development and improving cognition. In this study, the lipid differences between human MFGM and four dietary lipid sources (cow MFGM, soybean, krill, and yolk) were compared using the UHPLC-Q-Exactive MS-based lipidomics techniques. A total of 45 lipid classes and 5048 lipid species were detected. The analysis of phospholipid classes revealed that the lipid composition of human MFGM and cow MFGM was more similar than the other dietary-derived lipids. Additionally, the human MFGM lipid species were compared with cow MFGM, soybean, krill, and yolk, and 401, 416, 494, and 444 significantly different lipids were identified, respectively. Through lipid metabolic pathway analysis, differential lipids were mainly involved in the glycerophospholipid metabolic pathway. Overall, these results will provide a rationale for the future addition of lipids to infant formula to more closely approximate human MFGM lipid profiles. [ABSTRACT FROM AUTHOR]

Yak milk fat products constitute the base of Qinghai-Tibetan pastoralists’ daily food intake. Despite the great importance of fat in processing and pastoralists’ health, studies about yak milk fat are scarce. In this study, the lipid composition and the morphological properties of milk fat globule membranes (MFGMs) of yak milk were investigated. The results demonstrated that the yak milk had a higher cholesterol and sphingomyelin content compared to cow milk. In situ structural investigations performed at 25 °C by confocal microscopy showed the presence of lipid domains in yak MFGM, with a larger number and wider size range compared to cow milk. Moreover, the simultaneous localization of glycosylated molecules and polar lipids indicated that glycosylated molecules could be integrated into the lipid domains in yak MFGM. Different characteristics in yak MFGM could be related to the lipid composition and may affect the functions of yak milk lipids during processing and digestion. [ABSTRACT FROM AUTHOR]

• Infant formulae (IF) and human milk (HM) were comparatively analyzed. • Phospholipid species number was lower in IF (159) than in HM (231). • Phosphatidylethanolamine and sphingomyelin contents were lower in IF than in HM. • Soy lecithin and milk fat globule membrane supplements are promising for IF. • The particle size and potential of milk fat globule were smaller in IF than in HM. Phospholipids play key roles in infant nutrition and cognitive development. It is hypothesized that infant formula (IF) has lower phospholipid species, content and milk fat globule (MFG) structural integrity than human milk (HM). Herein, we performed qualitative and quantitative analyses of phospholipids in six classes of IF and HM using ultra-performance liquid chromatography with mass spectrometry. The contents of phosphatidylethanolamine (15.81 ± 7.20 mg/L) and sphingomyelin (35.84 ± 15.56 mg/L) in IF were significantly lower than those in HM (30.74 ± 17.38 mg/L, 45.53 ± 16.04 mg/L, respectively). Among the six IF classes, cow's milk-based IF had the highest number of phospholipid species, and IF containing milk fat globular membrane had the highest phospholipid content. The size, zeta potential, and amount of MFGs in IF were significantly lower than those in HM. These results may prove useful for designing better IF that mimic HM. [ABSTRACT FROM AUTHOR]

The miscibility of milk sphingomyelin (milk-SM) and cholesterol was investigated in this study. The effect of different physical states of milk-SM on its interactions with cholesterol was determined by the recording of isotherms of compression of Langmuir films for temperatures above and below the gel to Lα phase transition of milk-SM (T m ∼ 34 °C). For T = 15 °C < T m , the liquid expanded (LE) to liquid condensed (LC) phase transition of milk-SM monolayers was observed at surface pressures of 10–15 mN/m. For T = 43 °C > T m , the milk-SM molecules were in a LE phase regardless of the surface pressure applied. A phase diagram pressure – milk-SM/cholesterol composition was established. This study demonstrated that both temperature and surface pressure affected the miscibility between the milk-SM and cholesterol. The strongest attractive forces (i.e. condensing effect) were identified for 30 mol% cholesterol when the milk-SM was in the LE phase state. [ABSTRACT FROM AUTHOR]

The milk fat globule membrane (MFGM), which surrounds and stabilizes the fat globules, is released in buttermilk during cream churning. MFGM has many health benefits due to its composition rich in phospholipids and membrane proteins. Many techniques have been tried to separate the MFGM from the remaining milk solids non-fat, but they are challenging to carry out at an industrial scale. This research proposes a new approach to separating MFGM from buttermilk. This paper assessed the efficacy of hydroxyapatite (HA) cristal in interacting with MFGM isolates obtained from either raw or pasteurized cream. Different HA to MFGM ratios were used (10:1 and 20:1) to determine the impact of HA concentration on the adsorption. The results showed a very high affinity of the MFGM for HA and suggested the potential for its separation from buttermilk to improve its valorization. [ABSTRACT FROM AUTHOR]

• A comprehensive investigation of the effect of pasteurization on human milk proteins. • Hydrostatic high-pressure processed human milk was more closely resembled to raw milk. • Proteins in whey were less affected by pasteurization than proteins in other parts. The aim of this study was to comprehensively investigate the effect of Holder pasteurization (HoP) compared with that of hydrostatic high-pressure (HHP) processing on human milk proteins, including milk fat globule membrane (MFGM) proteins, whey proteins and caseins. Milk fat globules in milk processed by high-pressure were similar to those in raw milk in terms of their size distribution and microstructure, while the globules in milk processed by HoP were aggregated. The protein profiles of milk subjected to HHP processing more closely resembled those of raw milk than HoP milk. Proteins in milk whey were less affected by HoP or HHP than MFGM and casein proteins. The findings indicated a better preservation of the protein profile for HHP compared to HoP of human milk. [ABSTRACT FROM AUTHOR]

The bovine milk fat globule membrane (MFGM) has many associated biological activities, many of which are linked with specific carbohydrate structures of MFGM glycoconjugates. Bovine buttermilk is a commercially viable source of MFGM and is an under-valued by-product of butter making. However, the changes in buttermilk glycosylation over the course of lactation have not been extensively investigated. In this study, buttermilk was generated from three individual multiparous cows at 13 time points over the first three months of lactation. Buttermilk glycosylation was profiled using lectin microarrays and lectin blotting. Suggested differences in glycosylation, including N-glycosylation, sialylation and fucosylation, were observed between early and late time points and between individual animals. Overall, these data suggest temporal changes in the glycosylation of buttermilk proteins which may have an important impact on commercial isolation of glycosylated ingredients. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The microstructures of colostrum and mature bovine milk fat globule membrane (MFGM) were investigated using confocal laser scanning microscopy (CLSM) at different temperatures, and the relationships between microstructure variations and the chemical compositions of the MFGM were also examined. Using a fluorophore-labeled phospholipid probe, we found that non-fluorescent domains on the MFGM were positively correlated with the amount of sphingomyelin at both room (20 °C) and physiological (37 °C) temperatures. However, at the storage temperature (4 °C), there were more non-fluorescent domains on the MFGM. These results indicate that the heterogeneities in the MFGM are most likely to be the result of the lateral segregation of sphingomyelin at the room and physiological temperatures, and at the storage temperature, phospholipids with saturated fatty acids affect the formation of these domains. [ABSTRACT FROM AUTHOR]

Highlights: [•] We compared the in vivo intestinal digestion of raw and processed bovine creams. [•] We used the rat model and focused on interfacial structural changes. [•] The size of the fat globules increased in the stomach. [•] Interfacial spherical clusters appeared in chymes from pasteurised and raw cream. [•] Overall, there was a greater apparent digestion of pasteurised cream. [Copyright &y& Elsevier]

Highlights: [•] We compared the in vivo intestinal digestion of raw and processed bovine creams. [•] We used the rat model and focused on interfacial structural changes. [•] The formation of an interfacial liquid–crystalline lamellar phase was observed. [•] The apparition of crystals of possibly fatty acids and soaps was also observed. [•] There was a greater apparent digestion of pasteurised and homogenised cream. [Copyright &y& Elsevier]

Abstract: Milk fat globules are unique delivery systems for biologically active molecules in the gastrointestinal tract. However, their properties have not yet been fully investigated. In this study, we performed a comparative analysis of the polar lipid and fatty acid compositions of milk fat globules as a function of their size and investigated the structure of the milk fat globule membrane (MFGM). An optimised process of microfiltration was used to select the small milk fat globule (SMFG; 1.6μm) fractions and the large milk fat globule (LMFG; 6.6μm) fractions from the same initial whole milks (4.2μm). The SMFG-fractions contained significantly (i) higher amounts of polar lipids, 8.9±0.9 vs 2.7±0.3mg/g fat for LMFG-fractions and 6.3±0.5mg/g fat for whole milks, (ii) lower relative proportions of phosphatidylcholine and sphingomyelin in the MFGM, (iii) higher amounts of C12:0, C14:0, C16:0, C18:1 trans, C18:2 c9 tr11, and lower amounts of C18:0 and C18:1 c9 than did LMFG-fractions and whole milks. Whatever the size of native milk fat globules, the biophysical characterisation performed in-situ, using confocal laser scanning microscopy, showed heterogeneities in the MFGM. The lateral segregation of sphingomyelin in rigid liquid-ordered domains, surrounded by the fluid matrix of glycerophospholipids in the liquid-disordered phase, was revealed. The heterogeneous distribution of glycolipids and glycoproteins was also observed in the MFGM. A new model for the structure of the MFGM is proposed and discussed. The physical, chemical and biological consequences, (i) of the differences in milk fat globule compositions according to their size and (ii) of the specific structure of the MFGM due to sphingomyelin remain to be elucidated. [ABSTRACT FROM AUTHOR]

Abstract: Although buffalo milk is the second most produced milk in the world, and of primary nutritional importance in various parts of the world, few studies have focused on the physicochemical properties of buffalo milk fat globules. This study is a comparative analysis of buffalo and cow milk fat globules. The larger size of buffalo fat globules, 5 vs. 3.5μm, was related to the higher amount of fat in the buffalo milks: 73.4±9.9 vs. 41.3±3.7g/kg for cow milk. Buffalo milks contained significantly lower amount of polar lipids expressed per gram of lipids (0.26% vs. 0.36%), but significantly higher amount of polar lipids per litre of milk (+26%). Buffalo and cow milk fat globule membranes contain the same classes of polar lipids; phosphatidylethanolamine, sphingomyelin (SM) and phosphatidylcholine (PC) being the main constituents. A significant higher percentage of PC and lower percentage of SM were found for buffalo milks. The fatty acid analysis revealed that saturated fatty acids, mainly palmitic acid, trans fatty acids, linolenic acid (ω3) and conjugated linolenic acid were higher in buffalo milk than in cow milk. Such results will contribute to the improvement of the quality of buffalo milk-based dairy products. [Copyright &y& Elsevier]