Fat Globule Membrane Research Articles

Milk lipid and milk fat globule membrane production

The formation of the mammary secretion, proteins, lipids and ions is of fundamental importance to the dairy industry. The cellular interactions which allow secretion of protein and especially milk fat globules are still controversial. However, electron microscopy provides a basis for a straightforward scheme of secretion presented as a diagram. Each step in the process can then be defined, the published evidence for each assessed and areas which need further work clearly recognised. We also hope that the diagram will act as a way of combining global approaches such as proteomics or protein knockouts with the detail of electron microscopy.

Changes in milk fat globule membrane proteins along lactation stage of Laoshan dairy goat

The milk fat globule membrane (MFGM) is a complex structure with numerous functions, and its composition is affected by many factors. There have been few systematic investigations on goat MFGM proteome profiling during lactation. Individual milk samples from 15 healthy dairy goats were obtained at six lactation time points for investigation of the MFGM proteome using both data-independent acquisition (DIA) and data-dependent acquisition (DDA) proteomics techniques combined with multivariate statistical analysis. Using the DIA method, 890 variably abundant MFGM proteins were discovered throughout the lactation cycle. From 1 to 240 d, butyrophilin subfamily 1 member A1, lipoprotein lipase, perilipin-2, and adipose triglyceride lipase were upregulated, while APOE, complement C3, clusterin, and IgG were downregulated. Furthermore, from 1 to 90 d, annexin A1, annexin A2, and antithrombin-III were downregulated, then upregulated by d 240. Albumin had a high degree of connectedness, indicating that it was a key protein, according to protein–protein interaction research. Overall, our findings gave new insights into the biological features of MFGM protein in goat milk throughout lactation, which may aid in the creation of specialized MFGM products and infant formula.

The exploration of milk fat crystallization in milk fat globules by confocal Raman microscopy

Crystallization behavior within oil-in-water emulsion is a key factor for the properties and stability of many food products. Confocal Raman microscopy is a promising method to study such complex structures in situ. This study aimed at evaluating the feasibility of confocal Raman microscopy for visualizing milk fat crystallization and establishing a reliable data acquisition and processing methodology. Milk fat globules from raw milk were fixated in an agarose gel and crystallized at different temperatures. Confocal Raman microscopy was applied to collect two-dimensional area scans and supporting images were obtained by polarized light microscopy. The results revealed differences in lipid characteristics, crystal formation, and spatial distribution as a result of crystallization. Specific C-C stretching vibrations at 1063, 1083, and 1125 cm-1 were found to indicate lipid chain mobility and provide quantitative information on crystallinity. Additionally, the study successfully identified the triple-layered milk fat globule membrane. Different approaches for processing spectroscopic data were compared, emphasizing the importance of proper data handling. This novel spectroscopy imaging approach has significant potential in enhancing our understanding of structural heterogeneities of crystallized structures within complex food matrices.

Infant formula ingredients: Updates for clinicians.

Take Away Points All infant formulas meet specific nutritional standards, but there are now numerous versions of formula with newer ingredients that more closely approximate breast milk. Select infant formulas include ingredients such as human milk oligosaccharides, lactoferrin, or milk fat globule membranes, which may have additional clinical benefits. These ingredients are considered safe in infants but there is not dosing standardization across formula companies, and they may increase cost.

Preventative Effects of Milk Fat Globule Membrane Ingredients on DSS-Induced Mucosal Injury in Intestinal Epithelial Cells.

The goblet cells of the gastrointestinal tract (GIT) produce glycoproteins called mucins that form a protective barrier from digestive contents and external stimuli. Recent evidence suggests that the milk fat globule membrane (MFGM) and its milk phospholipid component (MPL) can benefit the GIT through improving barrier function. Our objective was to compare the effects of two digested MFGM ingredients with or without dextran sodium sulfate (DSS)-induced barrier stress on mucin proteins. Co-cultured Caco-2/HT29-MTX intestinal cells were treated with in vitro digests of 2%, 5%, and 10% (w/v) MFGM or MPL alone for 6 h or followed by challenge with 2.5% DSS (6 h). Transepithelial electrical resistance and fluorescein isothiocyanate (FITC)-dextran (FD4) permeability measurements were used to measure changes in barrier integrity. Mucin characterization was performed using a combination of slot blotting techniques for secreted (MUC5AC, MUC2) and transmembrane (MUC3A, MUC1) mucins, scanning electron microscopy (SEM), and periodic acid Schiff (PAS)/Alcian blue staining. Digested MFGM and MPL prevented a DSS-induced reduction in secreted mucins, which corresponded to the prevention of DSS-induced increases in FD4 permeability. SEM and PAS/Alcian blue staining showed similar visual trends for secreted mucin production. A predictive bioinformatic approach was also used to identify potential KEGG pathways involved in MFGM-mediated mucosal maintenance under colitis conditions. This preliminary in silico evidence, combined with our in vitro findings, suggests the role of MFGM in inducing repair and maintenance of the mucosal barrier.

Research on Ultrasonic Cavitation Treatment of Milk. An Experimental Approach

Homogenization is a standard milk processing method used to stabilize fat emulsions against gravitational separation. In the dairy industry, high-pressure homogenization is commonly applied, which requires significant energy input to achieve optimal processing conditions. The size and composition of fat globule membranes affect the sensory and rheological properties of secondary dairy products [1]. Recently, ultrasound has been frequently used for milk homogenization [2]. Unlike ultrasound, mechanical mixing does not provide emulsion stability even after prolonged treatment, highlighting the differences between these two technologies. These differences arise due to the varying degrees of fat globule and membrane disruption. However, dairy products with different physicochemical and functional properties can be produced using both ultrasonic treatment and in combination with other traditional homogenization methods.To study the impact of ultrasonic treatment parameters of milk, specifically their different values, on the size of fat inclusions and the stability of milk emulsion over a certain period after treatment.The study employed an experimental method of milk processing using an ultrasonic cavitation unit.The impact of ultrasonic cavitation on the quality of milk homogenization in a steady volume was investigated. Optimal process parameters, specifically the power of the ultrasonic field and treatment time, were experimentally determined. The use of ultrasonic frequency greater than 33 kHz was found to be impractical.It was experimentally proven that ultrasonic cavitation treatment of milk is extremely effective for obtaining homogeneous emulsions. Given that excessive heating (> 65 °C) is undesirable in the production of dairy products from a technological standpoint, a definite advantage of this method is the low-temperature (50 °C) mode compared to traditional processing methods (75–80 °C), which also positively affects the preservation of nutrients.

The affinity of milk fat globule membrane fragments and buttermilk proteins to hydroxyapatite

Buttermilk differs from skim milk by the presence of milk fat globule membrane (MFGM) fragments that are released during cream churning. MFGM is rich in health-promoting components, such as phospholipids and membrane proteins, but these compounds have a negative impact on buttermilk techno-functional properties in dairy applications. The isolation of MFGM from buttermilk improved its functionality while also recovering the MFGM bioactive components. Hydroxyapatite (HA) can be used to extract MFGM by adsorption via charged site interactions. However, the affinity of HA to MFGM or the main buttermilk proteins (casein micelles (CM), β-lactoglobulin (β-lg) and α-lactalbumin (α-lac)) is not known. The influence of important physicochemical parameters such as pH and temperature on these interactions is also unclear. For each buttermilk component, a quartz crystal microbalance diffusion analysis was performed to determine the maximum adsorption time and the attached mass density on HA-coated gold sensors. The influence of pH, ionic strength (IS), and temperature (T) on the affinity of each buttermilk component for HA particles was assessed using a 3-levels and 3-factors Box-Behnken design. The absorption rate was highest for the CM, followed by β-lg and α-lac, and then by the MFGM. Nevertheless, the final maximal attached mass densities to the HA were similar for the MFGM and CM, and 2.5 times higher than for β-lg and α-lac. This difference can be explained by the higher number of binding sites found in CM and their heavier mass. The model obtained by the Box-Behnken design plan showed that the adsorption of the CM changed with T, pH and IS. These results suggest that the techno-functional properties of buttermilk may be restored by specifically extracting MFGM with HA. Experiments are ongoing to determine conditions for fractionating MFGM directly from buttermilk.

Simulated in vitro infant digestion and lipidomic analysis to explore how the milk fat globule membrane modulates fat digestion

We hypothesized that the addition of milk fat globule membranes (MFGMs) to infant formula would improve its lipolysis, making it more similar to human milk (HM) and superior to commercial infant formula (CIF) in fat digestion. Therefore, we prepared two model infant formulas (MIFs) by adding MFGMs to dairy ingredients in different ways and compared their fat digestion behavior with those of HM and CIF. MFGMs were added alone (MIF1) and with other milk-based materials (MIF2) before homogenization. The addition of MFGMs reduced the flocculation of lipids and proteins in the gastric phase and promoted lipolysis in the intestine phase. The amount of free fatty acids released followed the order of HM > MIF1 > CIF ≥ MIF2. After digestion, the number of different glyceride species between each sample and HM reached 64 (MIF1), 73 (MIF2), 67 (CIF1), and 72 (CIF2). In conclusion, the fat digestion of MIF1 had the highest similarity with HM.

Phosphoproteomics Revealed Differentially Expressed Sites and Function of the Bovine Milk Fat Globule Membrane in Colostrum and Mature Milk.

One type of large and intricate post-translational modification of milk proteins that has significant biological implications is phosphorylation. The characterization of phosphoproteins found in the bovine milk fat globule membrane (MFGM) is still mostly unknown. Here, label-free phosphoproteomics was used to identify 94 phosphorylation sites from 54 MFGM phosphoproteins in bovine colostrum (BC) and 136 phosphorylation sites from 91 MFGM phosphoproteins in bovine mature milk (BM). αs1-Casein and β-casein were the most phosphorylated proteins in bovine colostrum. In bovine mature milk, perilipin-2 was the protein with the greatest number of phosphorylation sites. The results show that bovine colostrum MFGM phosphoproteins were mainly involved in immune function, whereas bovine mature MFGM phosphoproteins were mainly involved in metabolic function. Plasminogen and osteopontin were the most strongly interacting proteins in colostrum, whereas perilipin-2 was the most strongly interacting protein in bovine mature milk. This work demonstrates the unique alterations in the phosphorylation manner of the bovine MFGM protein during lactation and further expands our knowledge of the site characteristics of bovine MFGM phosphoproteins. This result confirms the value of MFGM as a reference ingredient for infant formula during different stages.

Preparation of milk fat globule membrane ingredients enriched in polar lipids: Composition characterization and digestive properties

In this study, milk fat globule membrane (MFGM) ingredients enriched in polar lipids was prepared using membrane filtration, including microfiltration, diafiltration, and ultrafiltration from butter serum powder. Polar lipids (phospholipids, sterols, and gangliosides) in prepared MFGM ingredients were analyzed by 31P NMR, GC-MS, and UPLC-MS/MS, respectively. The lipolysis degree and microstructure of MFGM ingredient and soybean lecithin-emulsification during in vitro digestion were also analyzed. Microfiltration showed higher concentration efficiency than ultrafiltration which increased 2.16% and 2.73% in phospholipid, respectively. Moreover, diafiltration concentrated more polar lipids (6.39% of phospholipid) than microfiltration. MFGM ingredients had high level of sphingomyelin (1.27-1.36%) and ration of GD3 than GM3 of 9.25-9.88-fold of total gangliosides. The different lipolysis behavior between MFGM ingredient-emulsification and soybean lecithin-emulsification were correlated with their different polar lipid compositions. Phospholipids from both MFGM ingredients and soybean lecithin could help maintain the initial structure during the gastric digestion. These results could provide a scientific basic for developing high-polar lipids food, particularly infant formulas and special functional foods.

Common proteins analysis of different mammals' mature milk by 4D-Label-Free

The milk proteins from samples of 13 different animals were identified utilizing 4D-Label-Free proteomics technology, leading to the identification of a substantial number of proteins. Among the various samples, Chinese people (CHP) milk proteins exhibited the highest count, with 1149 distinct proteins. Simultaneously, we identified common proteins present in these animal milk. It's notable presence in goat milk contributes to enhancing infant infection resistance, showcasing the beneficial role of lactoperoxidase. Galectin-3 binding protein (Gal-3BP) and tetraspanin in human milk are significantly higher than those in other animals, which determine the prominent antiviral effect of human milk and the important processes related to cell transduction. Furthermore, human milk, camel milk, goat milk and sheep milk proved to be rich sources of milk fat globule membrane (MFGM) proteins. The insights obtained from this study can serve as a foundational framework for exploring the role of different animal milk proteins in disease treatment and the composition of infant formula.

Formation by high power ultrasound of mimicking human fat emulsions stabilized with milk fat globule membrane

Ultrasound (US) is an innovative food processing technique that has gained significant interest for its role in modifying proteins. Our study focused on how different US time exposure influence the stability, interfacial structure, and rheological characteristics of whey protein infused with milk fat globule membrane (MFGM). Results indicated that a 9-min ultrasound treatment (US-9) of protein, yielding an emulsion with particle size of 338.3 nm and ζ-potential of −48 mV, was optimal for enhancing functional properties of the emulsion. The US-9 emulsion exhibited different surface rheological properties compared to other emulsions, revealing a heterogeneous MFGM stabilization interface predominantly made of MFGM fragments, free phospholipids, and proteins. The US-9 emulsion surface covered more phospholipids, indicating improved emulsion persistence, based on interfacial protein and phospholipid content calculations and microstructure observation. While the US-9 emulsion did not affect fat digestion, the US-6 min treatment resulted in an optimal fatty acid release rate. Further experimentation may be necessary to elucidate the intricate relationship between ultrasound duration and fat digestion. These findings have leaded to a better understanding of the emulsification capabilities of MFGM materials, as well as a foundation to improve infant formula quality.

Effects of maternal and perinatal factors on human milk fat globule membrane proteome: A data independent acquisition approach

Milk fat globule membrane proteins (MFGMP) are essential for supporting infants’ growth and development. The composition of MFGMP may vary depending on maternal and perinatal factors. However, there is a lack of research on the effects of parity, maternal age, and mode of delivery on MFGMP. This study attempts to address this knowledge gap by utilizing a data-independent acquisition (DIA) approach to perform proteomics comparisons of human MFGMP. Mature human milk (MHM) was collected from Chinese women with different parities (multiparous vs. primiparous, MP vs. PP), maternal reproductive ages (age ≥30 vs. age <30, MA1 vs. MA2), and delivery modes (cesarean delivery vs. vaginal delivery, CD vs. VD). We found variations in MFGMP between different cohorts. Differentially expressed proteins (DEP) in MHM of primiparous and multiparous women were mostly related to innate immune response which provides the first line of defense against pathogens. Higher abundances of these DEP were found in the MHM of multiparous women. Meanwhile, DEP in MHM of women with different delivery modes were primarily related to inflammatory response and innate immune response. We postulate that upregulation of the defense response-related DEP in MHM of women who had cesarean delivery was in relation to modulation of the cesarean delivery-induced inflammatory response and to strengthen the immune defense of newborns.

Identification and anti-oxidative potential of milk fat globule membrane (MFGM)-derived bioactive peptides released through in vitro gastrointestinal digestion

This study investigated the stability of milk fat globule membrane (MFGM) protein under simulated gastrointestinal conditions using an in vitro enzymatic digestion method. The optimal hydrolysis conditions were determined by monitoring the changes in particle size and zeta-potential of MFGM protein hydrolysates over time. Furthermore, the distribution of small molecular weight peptides with antioxidant activity was explored through DEAE-52 combined with in vitro cell experiments. Two novel antioxidant peptides (TGIIT and IITQ) were identified based on molecular docking technology and evaluated their potential scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS+) radicals. TGIIT and IITQ also demonstrated remarkable abilities in promoting mitochondrial biogenesis and activating Keap1/Nrf2 signaling pathway, which can effectively counteract skeletal muscle dysfunction induced by oxidative stress. Thus, MFGM-derived antioxidant peptides have the potential to be employed in food to regulate muscle protein metabolism and alleviate sarcopenia.

Structure, Biological Functions, Separation, Properties, and Potential Applications of Milk Fat Globule Membrane (MFGM): A Review.

The milk fat globule membrane (MFGM) is a thin film that exists within the milk emulsion, suspended on the surface of milk fat globules, and comprises a diverse array of bioactive components. Recent advancements in MFGM research have sparked a growing interest in its biological characteristics and health-related functions. Thorough exploration and utilization of MFGM as a significant bioactive constituent in milk emulsion can profoundly impact human health in a positive manner. Scope and approach: This review comprehensively examines the current progress in understanding the structure, composition, physicochemical properties, methods of separation and purification, and biological activity of MFGM. Additionally, it underscores the vast potential of MFGM in the development of additives and drug delivery systems, with a particular focus on harnessing the surface activity and stability of proteins and phospholipids present on the MFGM for the production of natural emulsifiers and drug encapsulation materials. MFGM harbors numerous active substances that possess diverse physiological functions, including the promotion of digestion, maintenance of the intestinal mucosal barrier, and facilitation of nerve development. Typically employed as a dietary supplement in infant formula, MFGM's exceptional surface activity has propelled its advancement toward becoming a natural emulsifier or encapsulation material. This surface activity is primarily derived from the amphiphilicity of polar lipids and the stability exhibited by highly glycosylated proteins.

Белки мембраны молочной жировой глобулы. 8. Белок, связывающий жирные кислоты (FABP)

This article continues a series of reviews about the structure and functions of milk fat globule membrane (MFGM) proteins in cow’s milk. This installment introduces publications on the biochemistry of fatty acid binding protein (FABP), which is a polypeptide component of MFGM. The article gives a brief history of its discovery, as well as profiles the proteins that bind hydrophobic ligands and describes the methods of FABP identification in MFGM preparations. Other aspects of FABP studies include: isoforms and modern nomenclature; amino acid sequence and reactions to post-translational modification; spatial organization; mechanisms for binding fatty acids to lipids; well-investigated and hypothesized physiological functions; the effect of haplotypes on milk fatty acids, yield, and protein content.

Evaluation of kefir grain microbiota, grain viability, and bioactivity from fermenting dairy processing by-products

Four dairy foods processing by-products (acid whey permeate (AWP), buttermilk (BM), sweet whey permeate (SWP), and sweet whey permeate with added milk fat globule ingredient (SWP+MFGM)) were fermented for 4 weeks and compared with traditional kefir milks for production of novel kefir-like dairy products. AU: Sweet whey permeates and SWP supplemented with 1.5% milk fat globule membrane (MFGM) showed to be the most viable by-products for kefir grain fermentation, exhibiting diverse abundance of traditional kefir microorganisms and positive indicators of bioactive properties. Grain viability was assessed with shotgun metagenomics, texture profile analysis, live cell counts, and scanning electron microscopy. Assessed bioactivities of the kefir-like products included antibacterial, antioxidant, potential anticancerogenic properties, and membrane barrier effect on human colorectal adenocarcinoma Caco-2 cells. All kefir grains were most abundant in Lactobacillus kefiranofaciens when analyzed with shotgun metagenomics. When analyzed with live cell counts on selective media, AWP kefir-like product had no countable Lactococcus spp. indicating suboptimal conditions for kefir grain microbiota survival and application for fermented dairy starter culture bacterium. Live cell counts were affirmed with kefir grain surface scanning electron microscopy images. SWP had the most adhesive kefir grain surface while SWP+MFGM had the largest exopolysaccharide (EPS) yield from grain extraction. All kefir and kefir-like products were able to achieve a 6-log reduction against Listeria innocua and Escherichia coli. Traditional milk kefirs had the highest antioxidant capacity for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay. AWP had a significantly higher DPPH antioxidant activity and SWP had the lowest Trolox equivalence concentration in the ABTS assay. Sweet whey and supplemented milk fat sweet whey had upregulation of Cldn-1 and Ocln-1 gene expression, which correspond with a significant increase in transepithelial electrical resistance.

Proteomic analysis of milk fat globule membrane proteins in mature human milk of women with and without gestational diabetes mellitus

Milk fat globule membrane proteins (MFGMP) in human milks have positive effects on infant’s health. As gestational diabetes mellitus (GDM) causes variations in MFGMP, it is essential to understand the effects of GDMon MFGMP. This study aims to investigate and compare the MFGMP (>3 months postpartum) of GDM and non-GDM (NGDM) women using four-dimensional-data-independent-acquisition proteomics technology. Principal component analysis shows significant differences in the MFGMP of GDM and NGDM women. A total of 4747 MFGMP were identified in maturehuman milk of GDM and NGDM women. Among these proteins, 174 differentially expressed proteins (DEPs) were identified in MFGM of GDM and NGDM women. Albumin (FC = 7.96) and transthyretin (FC = 2.57) which are related to insulin resistance and involved in thyroid hormone synthesis, are significantly up-regulated in MFGMP of GDM mothers indicating insulin resistance, imbalance of glucose homeostasis and poor glucose metabolism might persist in postpartum period.

Utilization of low-temperature heating method to improve skim milk production: Microstructure, stability, and constituents of milk fat globule membrane

In the process of defatting milk, preheating treatment is an important factor affecting the flavor of skim milk. Here, raw milk was preheated at different times and temperatures. Then laser confocal microscopy, multiple-light scattering instrument, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were used to analyze the microstructure of milk fat globule membrane (MFGM), milk stability, and MFGM protein (MFGMP) components. Results showed that phospholipid domain of MFGM changed from an ordered state (Lo) to a disordered state (Ld) with increase in treatment temperature and time, leading to an increase in MFGMP content in skim milk. During the stability test, the stability of raw milk decreased significantly with increase in preheating temperature, while the opposite was true for skim milk. Finally, the results of MFGMP differentiation analysis showed that, the content of ten taste-related MFGMPs in the control group samples was significantly lower compared to the optimal group (P < 0.05).

Supplemented Infant Formula and Human Breast Milk Show Similar Patterns in Modulating Infant Microbiota Composition and Function In Vitro.

The gut microbiota of healthy breastfed infants is often dominated by bifidobacteria. In an effort to mimic the microbiota of breastfed infants, modern formulas are fortified with bioactive and bifidogenic ingredients. These ingredients promote the optimal health and development of infants as well as the development of the infant microbiota. Here, we used INFOGEST and an in vitro batch fermentation model to investigate the gut health-promoting effects of a commercial infant formula supplemented with a blend containing docosahexaenoic acid (DHA) (20 mg/100 kcal), polydextrose and galactooligosaccharides (PDX/GOS) (4 g/L, 1:1 ratio), milk fat globule membrane (MFGM) (5 g/L), lactoferrin (0.6 g/L), and Bifidobacterium animalis subsp. lactis, BB-12 (BB-12) (106 CFU/g). Using fecal inoculates from three healthy infants, we assessed microbiota changes, the bifidogenic effect, and the short-chain fatty acid (SCFA) production of the supplemented test formula and compared those with data obtained from an unsupplemented base formula and from the breast milk control. Our results show that even after INFOGEST digestion of the formula, the supplemented formula can still maintain its bioactivity and modulate infants' microbiota composition, promote faster bifidobacterial growth, and stimulate production of SCFAs. Thus, it may be concluded that the test formula containing a bioactive blend promotes infant gut microbiota and SCFA profile to something similar, but not identical to those of breastfed infants.