갈락토올리고당과 생리활성 특성

Human milk is universally recognized as the optimal and most natural source of nutrition for newborns, offering benefits that extend far beyond basic energy and macronutrient provision. Among its complex constituents, human milk oligosaccharides (HMOs) represent the third most abundant solid component, surpassed only by lactose and lipids. HMOs are distinguished by their exceptionally high structural diversity—over 200 distinct structures have been identified to date. This structural complexity underlies the extensive biological functions HMOs perform within the infant’s body. HMOs play a pivotal role in promoting healthy growth, development, and overall well-being in infants and young children, functioning as indispensable bioactive molecules. Their key physiological activities include: immunomodulation and allergy prevention by promoting immune tolerance and reducing the risk of allergic diseases; potent anti-inflammatory and antioxidant effects that protect vulnerable infant tissues; support for brain development and cognitive enhancement through multiple mechanisms; anti-pathogenic properties, acting as soluble receptor analogs or “decoy” molecules to competitively block viral, bacterial, and other pathogen adhesion, thereby preventing colonization and infection in the gastrointestinal tract; and functioning as blood group substances. At the translational and application level, HMO research is actively driving cross-disciplinary innovation. Building on a deep understanding of their immunological and neurodevelopmental benefits, certain structurally defined HMOs have been successfully incorporated into infant formula. These HMO-supplemented formulas have received regulatory approval and are now commercially available worldwide, providing a nutritional alternative that more closely resembles human milk for infants who are not exclusively breastfed. This represents a significant step toward narrowing the compositional gap between formula and breast milk. Simultaneously, research into the symbiotic relationship between HMOs and the gut microbiota—particularly their role as selective prebiotic substrates promoting the growth of beneficial bacteria—has catalyzed the development of novel functional foods, dietary supplements, and microbiome-targeted therapies. These include advanced synbiotic formulations that combine specific probiotic strains with HMOs to synergistically optimize gut health and function. Furthermore, the intrinsic qualities of HMOs—including their natural origin, safety profile, biocompatibility, and proven antioxidant properties—have attracted growing interest in the emerging field of high-performance cosmetics. They are increasingly being explored as innovative functional ingredients in skincare products aimed at reducing oxidative stress and supporting skin health. This review aims to systematically synthesize recent advancements in HMO research, offering a comprehensive analysis centered on their complex composition and structural diversity; the molecular and cellular mechanisms underlying their diverse biological functions; their translational potential across sectors such as nutrition, medicine, and consumer care (including cosmetics); and the major challenges that persist in the field. It critically examines both foundational discoveries and recent breakthroughs. By integrating these interconnected themes, the review provides a holistic and up-to-date perspective on the scientific landscape of HMOs, highlighting their essential role in early-life nutrition and their expanding relevance across health and wellness applications. It also outlines promising directions for future research, with the goal of advancing evidence-based innovation in infant health and beyond.

Human milk oligosaccharides (HMOs) have garnered significant attention as one of the bioactive components in human milk, with growing applications in infant formula and food products. HMOs enhance butyrate production, which is produced by butyrate-producing bacteria such as Faecalibacterium prausnitzii and contributes to gut health through its diverse biological functions. However, the specific mechanisms by which individual HMOs promote butyrate production remain unclear. In this study, we conducted in vitro co-culture experiments of F. prausnitzii and Bifidobacterium bifidum, examining their relative abundance, fatty acid production, residual sugar levels, and gene expression. Our results revealed that B. bifidum utilizes HMOs and provides the constituent sugars to F. prausnitzii, thereby promoting butyrate production by F. prausnitzii. Furthermore, we found that the underlying mechanisms vary depending on the structure of the HMOs. Specifically, 2′-fucosyllactose and 3′-sialyllactose enhance the butyrate production efficiency of F. prausnitzii, while 6′-sialyllactose primarily promotes the growth of F. prausnitzii. These findings not only deepen our understanding of how HMOs influence infant gut health but also suggest new directions for developing nutritional products that leverage the distinct functional properties of each HMO.

A safety evaluation of mixed human milk oligosaccharides in rats

Chapter 2 - Structures, Classification, and Biosynthesis of Human Milk Oligosaccharides

How far is it from infant formula to human milk? A look at the human milk oligosaccharides

Background: Infant formula has a similar composition to breast milk, especially on key components like proteins, fats, and functional oligosaccharides. However, significant differences exist between bovine milk-based formula and human breast milk, particularly in the abundance and diversity of human milk oligosaccharides (HMOs). HMOs are pivotal in shaping the infant gut microbiome and immune system, with potential implications for infant health and development.Objectives: The literature review aimed to explore the role of HMOs in shaping the gut microbiome and strengthening the intestinal immune system in infants.Methods: Descriptive research was carried out in May-August 2023 using the literature review method. The design and implementation of this study referred to the guidelines outlined in the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) for Randomized Controlled Trials (RCTs). Literature sources were obtained from PubMed, Elsevier, Medline, Nature, PLOS, MPDI, JAMA, and Scopus publication databases with a limitation of the last ten years (2013-2023). One hundred twenty articles were obtained with these keywords and then selected in stages according to predetermined criteria. Six articles that met the criteria were retrieved from 2013-2023. The articles obtained were compiled, analyzed, and concluded by looking for similarities and dissimilarities, giving views, comparing, and summarizing.Results: The review revealed that HMOs significantly influenced the composition and function of the infant's intestinal microbiota, inhibiting harmful pathogens in the intestine. Therefore, HMOs as prebiotics were crucial in promoting intestinal immune system health in infants.Conclusion: Human Milk Oligosaccharides (HMOs) in infant formula substantially impacted the gut microbiome composition and the infant's intestinal immune system. Studies consistently showed that HMOs influenced the growth and diversity of gut bacteria, leading to outcomes similar to those seen in breastfed infants.

To investigate the relationship between human milk oligosaccharides(HMOs) levels in healthy mothers and infant physical growth. Based on a nationally representative, multi-stage stratified sampling cross-sectional survey on human milk composition, 195 lactating women within 6 months postpartum and their infants were enrolled. Human milk oligosaccharide(HMOs) concentrations were quantified using ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS). Infant anthropometric measurements(height, weight, and head circumference) were collected to calculate Z-scores. Associations between HMOs and infant growth were analyzed via general linear models. Total HMOs and total sialylated HMOs in mature breast milk showed negative correlations with weight for length Z-score(WLZ), weight for age Z-score(WAZ), and BMI for age Z-score(BAZ)(β=-0.80 to-0.25, P<0.05). Specifically, 6'-sialyllactosamine(6'-SL), sialyllacto-N-tetraose c(LSTc), and disialyllacto-N-tetraose(DSLNT) demonstrated significant negative associations with WAZ(β=-0.36 to-0.25, P<0.05). Total neutral non-fucosylated HMOs and lacto-N-tetraose(LNT) were also negatively correlated with length for age Z-score(LAZ) and WAZ(β=-0.33 to-0.25, P<0.05). Among secretor+/Lewis+(Se+Le+) mothers, the negative effects of total HMOs and sialylated HMOs on infant WLZ, WAZ, and BAZ were further strengthened(β=-1.22 to-0.48, P<0.05). Specific HMO concentrations in human milk during the first 6 months postpartum are associated with infant growth outcomes. Total HMOs, sialylated HMOs, and neutral non-fucosylated HMOs demonstrate negative associations with weight indicators.

Breast milk is rich in bioactive components, especially human milk oligosaccharides (HMOs), which are crucial for establishing gut microbiota. The 2'-FL (2-Fucosyllactose), one of the most abundant oligosaccharides in breast milk, functions as a selective prebiotic. Objective: To examine the effect of adding 2'-FL (2-Fucosyllactose) to an infant formula containing prebiotic galacto-oligosaccharides (GOSs) and fructo-oligosaccharides (FOSs) on the gut microbiome of healthy formula-fed infants. Methods: This study enrolled infants from three groups: an HMO experimental group (n = 29), a GOS/FOS control group (n = 30), and an exclusively breastfed (breast milk [BM]) reference group (n = 28). Fecal samples from the three groups in the first and fourth months of life were analyzed. The V3 and V4 regions of the 16S rRNA gene were amplified and sequenced on the Illumina MiSeq. ANOVA, Kruskal-Wallis, richness indices (Chao1, Shannon), UniFrac distances, and the Adonis tests were used to perform statistical analyses on the relative abundance of phyla and genera, as well as the alpha and beta-diversity of the gut microbiota. Results: After intervention, Actinobacteriota emerged as the predominant phylum in both the HMO (60.4%) and BM (46.6%) groups. Bifidobacterium and Escherichia-Shigella were identified as the two most abundant bacterial genera in both groups. Nevertheless, the statistical analysis showed that the relative abundance of Bifidobacterium in the HMO formula-fed group after intervention was similar to that in the BM group (p > 0.05). Infants in the HMO and GOS/FOS groups showed higher relative abundance of [Ruminococcus]_gnavus_group bacteria compared to those in the BM group. Groups fed with infant formula demonstrated higher alpha-diversity of gut microbiota compared to breastfed infants (p < 0.05), at the time of admission as well as after the intervention. Beta-diversity was significantly different among the three groups, according to type of feeding. Infants fed a 2'-FL-supplemented infant formula exhibited growth comparable to that of breastfed infants throughout the intervention period, demonstrating that the formula was both safe and well tolerated. Conclusions: Adding 2'-FL to an infant formula containing 4 g/L of GOS + FOS resulted in a stronger bifidogenic effect compared to the formula without 2'-FL.

Breastfeeding is integral in the proper maturation of the intestinal barrier and protection against inflammatory diseases. When human milk (HM) is not available, supplementation with HM bioactives like Human Milk Oligosaccharides (HMOs) may help in providing breastfeeding barrier-protective benefits. An increasing HMO variety is becoming industrially available, enabling approaching the HMO complexity in HM. We aimed at assessing the impact of blends of available HMOs on epithelial barrier function in vitro. The capacity of individual [2′-Fucosyllactose (2′FL), Difucosyllactose, Lacto-N-tetraose, Lacto-N-neotetraose, 3′-Siallylactose and 6′-Siallylactose] or varying combinations of 3, 5 and 6 HMOs to modulate fluorescein-isothiocyanate (FITC)-labelled Dextran 4 KDa (FD4) translocation and/or transepithelial resistance (TEER) was characterized in Caco-2: HT29- methotrexate (MTX) cell line monolayers before and after an inflammatory challenge with TNF-α and IFN-γ. The six HMO blend (HMO6) dose-dependently limited the cytokine-induced FD4 translocation and TEER decrease and increased TEER values before challenge. Similarly, 3 and 5 HMO blends conferred a significant protection against the challenge, with 2′FL, one of the most abundant but most variable oligosaccharides in HM, being a key contributor. Overall, our results suggest differential ability of specific HMOs in modulating the intestinal barrier and support the potential of supplementation with combinations of available HMOs to promote gut health and protect against intestinal inflammatory disorders.

Human milk oligosaccharides (HMOs) represent the thirdmost abundantfraction of biomolecules in human milk (HM) and play a crucial rolein infant health and development. The unique contributions of HMOsto healthy development of breast-fed infants are assumed to rely onthe extraordinary complexity and diversity of HMO isomeric structures,which in turn still cause a huge analytical challenge. Many contemporaryanalytical methods aiming for more detailed HMO characterization combineion mobility (IM) with LC-MS for enhanced structural resolution butare typically lacking the robustness necessary for application toHM cohorts with hundreds of samples. To overcome these challenges,we introduce a novel, robust all-ion fragmentation (AIF) LC-ESI-IM-MSmethod integrating four analytical dimensions: high-resolution LCseparation, IM drift time, accurate mass precursor, and fragment ionmeasurements. This four-dimensional (4D) analytical characterizationis sufficient for resolving various HMO structural isomers in an efficientway. Thereby, up to 200 HMO compounds with a maximum degree of polymerizationof 13 could be simultaneously identified and relatively quantified.We devised two methods using this 4D analytical approach. One intendedfor in-depth characterization of multiple known but also novel HMOstructures and the second is designed for robust, increased-throughputanalyses. With the first approach, five trifucosyl-lacto-N-tetraose isomers (TF-LNTs), four of which were never detected beforein HM, as well as additional difucosyl-lacto-N-heaoseisomers (DF-LNHs), were revealed and structures fully elucidated byAIF and IM. This exemplifies the potential of our method for in-depthcharacterization of novel complex HMO structures. Furthermore, theincreased-throughput method featuring a shorter LC gradient was appliedto real-world HM samples. Here, we could differentiate the HM typesI–IV based on a broader range of partly new marker HMOs. Wecould also derive valuable new insights into variations of multipleand rare HMOs up to DP 11 across lactational stages. Overall, ourAIF LC-ESI-IM-MS approach facilitates in-depth monitoring and confidentidentification of a broad array of distinct and simple to very complexHMOs. We envision this robust AIF LC-ESI-IM-MS approach to advanceHMO research by facilitating the characterization of a broad rangeof HMOs in high numbers of HM samples. This may help to further extendour understanding about HMOs structure–function relationshipsrelevant for infants’ healthy development

Background/Objectives: HMOs are the third most abundant solid component after lactose and fats in human milk. This study aimed to examine the relationships between maternal diet and HMO composition and concentration in human milk among lactating women in Latvia. Methods: Pooled 24 h human milk samples, 72 h food diaries, and questionnaires on anthropometric and sociodemographic characteristics were collected from 68 exclusively breastfeeding women residing in Latvia. HMOs were analyzed by UHPLC/FLD, and dietary data were analyzed using the Estonian NutriData program. Results: The eight most abundant HMO structures were determined with total concentration ranging between 178.66 and 32,910.09 mg L−1. 2′-FL was the most prevalent HMO in human milk (median concentration—3647 mg L−1), followed by 3′-FL (1436.74 mg L−1). Participants had an insufficient intake of vegetables, fruits, berries, milk and dairy products, and fish, leading to vitamin A, vitamin C, folate, and iodine intakes lower than recommended for lactating women. Limitation or exclusion of milk and dairy products from the diet was associated with a higher 2′-FL concentration in human milk (p = 0.037). Preference for “zero sugar” products was associated with a higher 3′-FL, 6′-GL, LNnT, 6′-SL, LNDFH II concentration in human milk (p < 0.050). Dietary supplement use (e.g., vitamin D, calcium) was also associated with differences in HMO composition and concentration in milk (p < 0.050). Conclusions: The findings highlight the importance of dietary habits and supplement use in shaping HMO profiles, though more human milk samples and dietary data need to be evaluated to draw further conclusions.

The characteristic composition of the intestinal microbiota of breast-fed neonates is due to the presence of particular substances in human milk. This chapter aims to identify the components that are able to promote the prebiotic effect in human milk, and is oriented towards modifying the composition of infant formulas in order to obtain an intestinal microbiota similar to that of breast-fed babies. It discusses history, synthesis, structure, metabolism of human milk oligosaccharides (HMO). No natural substances have the same biochemical composition as that of HMO, nor can they be synthesized in large quantities at acceptable prices. To overcome these problems, the industry has focused on the production of several carbohydrates, so-called nondigestible oligosaccharides (NDO), which, although having compositions different from those of HMO, are able to selectively stimulate the growth of bifidobacteria and lactobacilli in the colon, reproducing the prebiotic effects of HMO. In conclusion, data related to the use of NDO in infant formulas clearly show the efficacy of galactooligosaccharides (GOS)/inulin mixtures, the capacity of GOS to induce a bifidogenic effect, even if in only a small number of studies, and the positive effects of FOS at a concentration of 4.0 g/liter on the intestinal microbiota. It has been demonstrated that, due to the presence of resistant bonds in their molecules, NDO are able to exert a prebiotic effect. This further confirms that, because of their peculiar structure, HMO have a very significant role in modulating the intestinal microbiota of neonates.

Background: Human milk is a complex fluid comprised of myriad substances, with one of the most abundant substances being a group of complex carbohydrates referred to as human milk oligosaccharides (HMOs). There has been some evidence that HMO profiles differ in populations, but few studies have rigorously explored this variability.Objectives: We tested the hypothesis that HMO profiles differ in diverse populations of healthy women. Next, we examined relations between HMO and maternal anthropometric and reproductive indexes and indirectly examined whether differences were likely related to genetic or environmental variations.Design: In this cross-sectional, observational study, milk was collected from a total of 410 healthy, breastfeeding women in 11 international cohorts and analyzed for HMOs by using high-performance liquid chromatography.Results: There was an effect of the cohort (P < 0.05) on concentrations of almost all HMOs. For instance, the mean 3-fucosyllactose concentration was >4 times higher in milk collected in Sweden than in milk collected in rural Gambia (mean ± SEM: 473 ± 55 compared with 103 ± 16 nmol/mL, respectively; P < 0.05), and disialyllacto-N-tetraose (DSLNT) concentrations ranged from 216 ± 14 nmol/mL (in Sweden) to 870 ± 68 nmol/mL (in rural Gambia) (P < 0.05). Maternal age, time postpartum, weight, and body mass index were all correlated with several HMOs, and multiple differences in HMOs [e.g., lacto-N-neotetrose and DSLNT] were shown between ethnically similar (and likely genetically similar) populations who were living in different locations, which suggests that the environment may play a role in regulating the synthesis of HMOs.Conclusions: The results of this study support our hypothesis that normal HMO concentrations and profiles vary geographically, even in healthy women. Targeted genomic analyses are required to determine whether these differences are due at least in part to genetic variation. A careful examination of sociocultural, behavioral, and environmental factors is needed to determine their roles in this regard. This study was registered at clinicaltrials.gov as NCT02670278.

Human milk oligosaccharides (HMO) are one of the major components of human milk. HMO are non-digestible by the human gut, where they are known to play important functions as prebiotics and decoys for binding pathogens. Moreover, it has been proposed that HMO may provide sialic acids to the infant that are important in brain development, however this would require absorption of HMO into the bloodstream. HMO have consistently been found in the urine of humans and other mammals, suggesting systemic absorption. Here, we present a procedure for the profiling of milk oligosaccharides (MO) in plasma samples obtained from 13 term infants hospitalized for surgery for congenital heart disease. The method comprises protein denaturation, oligosaccharide reduction, and porous graphitized carbon solid phase extraction for purification followed by analysis using nHPLC-PGC-chip-TOF-MS. Approximately 15 free MO were typically observed in the plasma of human infants, including LNT, LDFP, LNFT, 3'SL, 6'SL, 3'SLN, and 6'SLN, of which the presence was confirmed using fragmentation studies. A novel third isomer of SLN, not found in human or bovine milk was also consistently detected. Differences in the free MO profiles were observed between infants that were totally formula-fed and infants that received at least some part breast milk. Our results indicate that free MO similar in structure to those found in human milk and urine are present in the blood of infants. The method and results presented here will facilitate further research toward the possible roles of free MO in the development of the infant.

Human milk oligosaccharides (HMOs) are an evolutionarily significant advantage bestowed by mothers for facilitating the development of the infant's gut microbiota. They can avoid absorption in the stomach and small intestine, reaching the colon successfully, where they engage in close interactions with gut microbes. This process also enables HMOs to exert additional prebiotic effects, including regulating the mucus layer, promoting physical growth and brain development, as well as preventing and mitigating conditions such as NEC, allergies, and diarrhea. Here, we comprehensively review the primary ways by which gut microbiota, including Bifidobacteria and other genera, utilize HMOs, and we classify them into five central pathways. Furthermore, we emphasize the metabolic benefits of bacteria consuming HMOs, particularly the recently identified intrinsic link between HMOs and the metabolic conversion of tryptophan to indole and its derivatives. We also examine the extensive probiotic roles of HMOs and their recent research advancements, specifically concentrating on the unsummarized role of HMOs in regulating the mucus layer, where their interaction with the gut microbiota becomes crucial. Additionally, we delve into the principal tools used for functional mining of new HMOs. In conclusion, our study presents a thorough analysis of the interaction mechanism between HMOs and gut microbiota, emphasizing the cooperative utilization of HMOs by gut microbiota, and provides an overview of the subsequent probiotic effects of this interaction. This review provides new insights into the interaction of HMOs with the gut microbiota, which will inform the mechanisms by which HMOs function.