Search

Human milk is universally recognized as the preferred food for infants during the first 6 mo of life because it provides not only essential and conditionally essential nutrients in necessary amounts but also other biologically active components that are instrumental in protecting, communicating important information to support, and promoting optimal development and growth in infants. Despite decades of research, however, the multifaceted impacts of human milk consumption on infant health are far from understood on a biological or physiological basis. Reasons for this lack of comprehensive knowledge of human milk functions are numerous, including the fact that milk components tend to be studied in isolation, although there is reason to believe that they interact. In addition, milk composition can vary greatly within an individual as well as within and among populations. The objective of this working group within the Breastmilk Ecology: Genesis of Infant Nutrition (BEGIN) Project was to provide an overview of human milk composition, factors impacting its variation, and how its components may function to coordinately nourish, protect, and communicate complex information to the recipient infant. Moreover, we discuss the ways whereby milk components might interact such that the benefits of an intact milk matrix are greater than the sum of its parts. We then apply several examples to illustrate how milk is better thought of as a biological system rather than a more simplistic "mixture" of independent components to synergistically support optimal infant health.

Partial digestion of milk proteins leads to the formation of numerous bioactive peptides. Previously, our research team thoroughly examined the decades of existing literature on milk bioactive peptides across species to construct the milk bioactive peptide database (MBPDB). Herein, we provide a comprehensive update to the data within the MBPDB and a review of the current state of research for each functional category from in vitro to animal and clinical studies, including angiotensin-converting enzyme (ACE)-inhibitory, antimicrobial, antioxidant, dipeptidyl peptidase (DPP)-IV inhibitory, opioid, anti-inflammatory, immunomodulatory, calcium absorption and bone health and anticancer activity. This information will help drive future research on the bioactivities of milk peptides. [ABSTRACT FROM AUTHOR]

Whey protein concentrate (WPC) is consumed for its high protein content. The structure and biological functionality of whey proteins in WPC powders may be affected by the drying technique applied. However, the specific impact of spray drying and freeze drying on the overall protein profile of whey protein derived from sweet whey streams at scale is unknown. Herein, we examine the effects of commercial‐scale freeze drying and spray drying on WPC to determine which method better preserves bioactive whey proteins, with the goal of helping the dairy industry create high‐value products that meet the growing consumer demand for functional dairy products. WPCs were produced from pasteurized liquid whey using either a commercial spray dryer or freeze dryer. A variety of analytical techniques, including enzyme‐linked immunosorbent assay, polyacrylamide gel electrophoresis, and bottom‐up proteomics using liquid chromatography‐tandem mass spectroscopy were used to identify, quantify, and compare the retention of bioactive proteins in WPC before and after spray drying and freeze drying. In addition, the extent of denaturation was studied via solubility testing, differential scanning calorimetry, and hydrophobicity assessment. There was little to no difference in the retention or denaturation of key bioactive proteins between spray‐dried and freeze‐dried WPC powders. There was a higher percentage of select Maillard modifications in freeze‐dried and spray‐dried powders than in the control. The lack of significant differences between spray drying and freeze drying identified herein indicates that freeze drying does not meaningfully improve retention of bioactive proteins compared with spray drying when performed after multiple pasteurization steps. Practical Application: This study aimed to provide insight into the impacts of spray drying versus freeze drying on whey proteins. Overall, our results indicate that for commercial dairy processing that involves multiple rounds of pasteurization, freeze drying does not meaningfully improve the retention of bioactive proteins compared with spray drying. These findings may help the food and dairy industry make informed decisions regarding the processing of its whey protein products to optimize nutritional value. [ABSTRACT FROM AUTHOR]

Lactoferrin (LF) is a major component of human milk. LF supplementation (currently bovine) supports the immune system and helps maintain iron homeostasis in adults. No recombinant human lactoferrin (rhLF) is available for commercial food use. To determine the extent to which rhLF (Effera™) produced by Komagataella phaffii digests similarly to hmLF, a validated in vitro digestion protocol was carried out. Bovine LF (bLF) was used as an additional control, as it is approved for use in various food categories. This study compared the extent of intact protein retention and the profile of peptides released in hmLF, bLF and rhLF (each with low and high iron saturation) across simulated adult gastric and intestinal digestion using gel electrophoresis, ELISA and LC-MS. Intact LF retention across digestion was similar across LF types, but the highest iron-saturated hmLF had greater retention in the simulated gastric fluid than all other sample types. Peptides identified in digested hmLF samples strongly correlated with digested rhLF samples (0.86 < r < 0.92 in the gastric phase and 0.63 < r < 0.70 in the intestinal phase), whereas digested bLF samples were significantly different. These findings support the potential for rhLF as a food ingredient for human consumption. [ABSTRACT FROM AUTHOR]

Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In this study, we applied mass spectrometrybased peptidomics to characterize peptides present in colostrum before and after in vitro digestion. Sequence-based computational modeling was applied to predict peptides with antimicrobial activity. We identified more peptides in undigested samples, yet the abundances were much higher in the digested samples. Heatmapping demonstrated highly different peptide profiles between undigested and digested samples. Four peptides (αS1-casein [157–163], αS1-casein [157–165], β-casein [153–159] and plasminogen [591–597]) were selected, synthesized and tested against common pathogenic bacteria associated with necrotizing enterocolitis. All four exhibited bacteriostatic, though not bactericidal, activities against Klebsiella aerogenes, Citrobacter freundii and Serratia marcescens, but not Escherichia coli. [ABSTRACT FROM AUTHOR]

Background: Although human‐milk feeding reduces the risk of necrotizing enterocolitis (NEC) in preterm infants compared with formula feeding, the exact risk‐reduction mechanism remains unknown. As NEC occurs at the distal small intestine in which digestion has occurred, we applied proteomics to examine the extent to which colostrum proteins survive simulated infant in vitro–digestion and, thus, have potential to exert biological function. Methods: Ten preterm colostrum samples were left undigested or in vitro–digested, and lipopolysaccharide (LPS)‐binding protein, soluble cluster of differentiation 14, and tumor necrosis factor (TNF) receptors I and II were measured using enzyme‐linked immunosorbent assay in all undigested and in vitro–digested samples. Fully differentiated Caco‐2 cells were exposed to digested colostrum samples before stimulation with LPS or TNF or no stimulation. Inflammation (interleukin‐8) and cytotoxicity (lactate dehydrogenase) were measured. Proteomic analyses of undigested and in vitro–digested samples were done using mass spectrometry. Results: We found that most proteins in colostrum are significantly, if not completely, degraded after in vitro–digestion. We found select individual and combination digestion‐resistant proteins that were positively correlated with LPS‐ and TNF‐induced inflammation. Conclusion: These results indicate the importance of considering the extent to which specific dietary compounds survive digestion to reach their site of claimed action (distal intestine) and that some digestion‐resistant proteins may be contributing toward "low‐grade" inflammation that is necessary to promote intestinal growth and maturation during early infancy. This work provides the most detailed understanding of human‐milk protein degradation with simulated infant in vitro–digestion to date. [ABSTRACT FROM AUTHOR]

High-pressure processing (HPP) of donor human milk (DM) minimally impacts the concentration and bioactivity of some important bioactive proteins including lactoferrin, and bile salt-stimulated lipase (BSSL) compared to Holder pasteurization (HoP), yet the impact of HPP and subsequent digestion on the full array of proteins detectable by proteomics remains unclear. We investigated how HPP impacts undigested proteins in DM post-processing and across digestion by proteomic analysis. Each pool of milk (n = 3) remained raw, or was treated by HPP (500 MPa, 10 min) or HoP (62.5 °C, 30 min), and underwent dynamic in vitro digestion simulating the preterm infant. In the meal, major proteins were minimally changed post-processing. HPP-treated milk proteins better resisted proximal digestion (except for immunoglobulins, jejunum 180 min) and the extent of undigested proteins after gastric digestion of major proteins in HPP-treated milk was more similar to raw (e.g., BSSL, lactoferrin, macrophage-receptor-1, CD14, complement-c3/c4, xanthine dehydrogenase) than HoP. • Study assessing how human milk pasteurization impacts protein digestion in vitro. • Pasteurization minimally changes proteins (count, abundance) pre-digestion. • Increased proteolysis in Holder vs. high-pressure milk during gastric phase. • Simulated digestion of raw and high-pressure milk yields similar profiles. • Key proteins in high-pressure milk better resist proximal digestion vs. Holder. [ABSTRACT FROM AUTHOR]

Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In this study, we applied mass spectrometry-based peptidomics to characterize peptides present in colostrum before and after in vitro digestion. Sequence-based computational modeling was applied to predict peptides with antimicrobial activity. We identified more peptides in undigested samples, yet the abundances were much higher in the digested samples. Heatmapping demonstrated highly different peptide profiles between undigested and digested samples. Four peptides (α S1 -casein [157-163], α S1 -casein [157-165], β-casein [153-159] and plasminogen [591-597]) were selected, synthesized and tested against common pathogenic bacteria associated with necrotizing enterocolitis. All four exhibited bacteriostatic, though not bactericidal, activities against Klebsiella aerogenes , Citrobacter freundii and Serratia marcescens , but not Escherichia coli .