GOS (Galactooligosaccharides)

In 2011, the Committee on Nutrition of the European Society for Paediatric Gastroenterology, Hepatology and Nutrition systematically reviewed published evidence related to the safety and health effects of the administration of formulae supplemented with pro- and/or prebiotics compared with unsupplemented formulae. We updated evidence on the effects of the administration of prebiotic-supplemented infant formulae (IF) compared with unsupplemented IF. Five databases were searched up to March 2017 for randomised controlled trials. In all, forty-one publications were identified, including twenty-five new publications. The administration of currently evaluated prebiotic-supplemented formulae to healthy infants does not raise safety concerns with regard to growth and adverse effects. Some favourable clinical effects are possible, primarily stool softening, which may be beneficial in some infants. Currently, there is no existing robust evidence to recommend the routine use of prebiotic-supplemented formulae. The latter conclusion may reflect the small amount of data on specific prebiotics and outcomes, rather than a genuine lack of an effect. The efficacy and safety should be considered for each prebiotic(s)-supplemented formula.

Probiotic supplements containing high β-galactosidase-producing bacteria may aid in the management of lactose intolerance. We previously isolated a strain of Bifidobacterium adolescentis, iVS-1, from the fecal sample of a human donor after consumption of galactooligosaccharides (GOS), a prebiotic derived from lactose. Therefore, it was hypothesized that iVS-1 might reduce symptoms associated with lactose maldigestion. Compared to other probiotic strains, iVS-1 had high β-galactosidase activity and reduced gas formation by fecal communities during in vitro fermentations of lactose or milk. A randomized placebo-controlled clinical trial was then conducted with 21 lactose maldigesters, randomized to receive either B. adolescentis iVS-1 (n = 11) or placebo (n = 10) daily for 2 weeks. Compared to the two-week run-in period, iVS-1 abundance was higher both at the end of the treatment period (p = 0.0005) and after the 2-week post-treatment period (p = 0.045). The iVS-1 group reported less overall daily symptoms during the treatment period when compared to placebo (p = 0.032) and had significant improvement for fecal urgency (p = 0.033) and diarrhea (p = 0.006). The metabolism of lactose, reduction of gas, and improvement of multiple gastrointestinal symptoms suggest that B. adolescentis iVS-1 may be an effective treatment for lactose intolerance. Trial Registration: The trial is registered at ClinicalTrials.gov (https://clinicaltrials.gov/study/NCT05668468). © 2025 The Author(s). Published with license by Taylor & Francis Group, LLC.

BACKGROUND/OBJECTIVES: Prebiotics, which are non-digestible compounds that selectively modulate gut microbiota, are recognized for their potential to promote host health. Although their bifidogenic effect is well documented, a systematic synthesis of how this microbial modulation translates into clinical gastrointestinal (GI) and metabolic outcomes across diverse populations is needed. This review aims to integrate mechanistic insights with clinical evidence to elucidate the pathway from prebiotic structures to tangible health benefits. METHODS: This comprehensive narrative review details the structural properties of major prebiotics (e.g., inulin, FOS, and GOS) that govern their fermentation and the production of short-chain fatty acids (SCFAs). To evaluate clinical efficacy, an analysis of 22 randomized controlled trials from the past decade was conducted, focusing on human studies that utilized ISAPP-recognized prebiotics as the sole intervention. RESULTS: The analysis confirms that prebiotic supplementation consistently increased the abundance of beneficial bacteria (e.g., Bifidobacterium and Lactobacillus) and SCFA production. These changes are associated with significant clinical improvements, including enhanced stool frequency and consistency, strengthened intestinal barrier function, and modulated immune responses. Benefits have been documented in healthy individuals, children, the elderly, and those with conditions such as constipation, metabolic syndrome, and antibiotic-associated dysbiosis. However, significant inter-individual variability in response was evident, and the study designs showed notable heterogeneity in prebiotic type, dosage, and duration. CONCLUSIONS: Prebiotics are effective modulators of gut health, driving clinical benefits through selective microbial fermentation and SCFA production. The documented heterogeneity and variability highlight the need for future research to focus on personalized nutritional strategies. Key priorities include standardizing intervention protocols, elucidating dose-response relationships, integrating multi-omics data to link taxonomy to function, and exploring novel applications such as synbiotic formulati

BACKGROUND: Partially hydrolyzed cow's milk protein (PHP) formulas are nutritionally complete and have a high-quality protein composition, and extensive history of safe use. The current study evaluated growth and safety in healthy term infants receiving a new PHP formula with an added prebiotic blend. METHODS: In this multi-center, double-blind, controlled, parallel, prospective study, healthy term infants were randomized to receive one of two formulas through 365 days of age: previously marketed intact cow's milk protein formula (Control, n = 122) or investigational PHP formula (INV-PHP, n = 122). Both formulas had an added prebiotic blend of polydextrose (PDX) and galactooligosaccharides (GOS) (1:1, 4 g/L). The primary outcome was rate of weight gain (g/day) from 14 to 120 days of age. To establish equivalence between study formulas, the 90% two-sided confidence interval (CI) of the mean group difference in body weight growth rate from 14 to 120 days of age needed to be contained within a predefined equivalence interval (± 3 g/day). Growth rates through Day 120 and achieved anthropometrics through Day 365 were analyzed by ANOVA. Parent-reported tolerance outcomes were also collected. Medically confirmed adverse events were collected throughout the study period. RESULTS: Of 244 infants enrolled and randomized (Control, n = 122; INV-PHP, n = 122); 175 completed study feeding through Day 120 (Control, n = 91; INV-PHP, n = 84). Equivalence in rate of weight gain from 14 to 120 days of age was demonstrated with the difference in means of 0.5 g/day and 90% CI [- 1.10, 2.08 g/day] within the predefined equivalence interval (± 3 g/day). Mean achieved weight remained between 25th -75th reference percentiles of the WHO growth standard through Day 180 by sex and subsequently tracked between 50th -90th percentiles through Day 365. Formula acceptance and tolerance were good. Stool consistency remained soft in both groups throughout the study. No significant group differences in mean fussiness and gassiness scores, or medically confirmed adverse events were detected. A total of 159 participants completed the Day 365 visit (Control, n = 82; INV-PHP, n = 77). CONCLUSIONS: Ove

In this randomized, double-blind controlled trial, 182 Filipino children aged 2-3 years received either an experimental young child formula (EYCF) containing a combination of Limosilactobacillus reuteri DSM 17938 and galacto-oligosaccharides (GOS; n = 91) or a minimally fortified milk (CM; n = 91) for 6 months. Primary outcome was tibia speed of sound and secondary outcomes were muscle strength, blood vitamin D levels, bone turnover markers, gut microbiota, fecal calcium fatty acid soaps and gastro-intestinal tolerance. Compared to CM, those in the EYCF group showed increased tibia speed of sound after 3 and 6 months. The intervention remodeled the stool microbiome composition, assessed by shotgun metagenomics, with enrichment of L. reuteri and higher bifidobacteria presence in the EYCF group. Increased L. reuteri abundance after 6 months of EYCF consumption associates with higher bone quality and muscle strength. Stool metabolomics show 45 metabolites modulated by EYCF consumption and associated to microbiome compositional changes, leading to enrichment of tryptophane and indole metabolism. In summary, consumption of EYCF containing a L. reuteri + GOS synbiotic improves musculoskeletal development in toddlers via modulation of microbiota composition and function. These results provide insights on gut-musculoskeletal crosstalk during early life. Clinicaltrial.gov NCT04799028. © 2025. The Author(s).

BACKGROUND: New primary preventive therapeutic strategies for atopic dermatitis (AD) are needed. Atopic diseases are associated with disrupted gut microbial balance in early life, suggesting that optimizing microflora through intervention could improve health. Prebiotics, which are immunomodulatory sugars, promote gut microbiota diversity. Most clinical trials focus on improving postnatal infant gut colonization, but prenatal life is crucial for establishing tolerance mechanisms. Preclinical studies indicate that maternal intake of galacto-oligosaccharide (GOS)/inulin prebiotics decreases the risk of food allergy in offspring. OBJECTIVES: To determine whether antenatal prebiotic intake prevents AD in children at high risk of atopy. METHODS: The multicentre double-blind randomized PREGRALL clinical trial ran from February 2018 to April 2023 (ClinicalTrials.gov NCT03183440). The follow-up period extended from 20 weeks of amenorrhoea in pregnant women to their infants reaching 1 year of age. Women with a physician-diagnosed history of atopy (asthma, allergic rhinitis, AD or food allergy) were selected for inclusion. The women were randomized to daily prebiotic GOS/inulin (n = 188) or placebo (maltodextrin; n = 188 participants) intake from 20 weeks' gestation until delivery. The first outcome was the occurrence of AD at 1 year in children at risk of the disease. Secondary endpoints included AD severity, quality of life, prebiotic tolerance and the prevalence of other atopic diseases. RESULTS: Of 376 pregnant women included in the trial, prebiotic supplementation did not prevent AD at 1 year (intention-to-treat population odds ratio 1.01, 95% confidence interval 0.59-1.74; P = 0.97) or reduce disease severity in their children. Subgroup analyses by breastfeeding status or delivery mode revealed no differences. No effects on allergen sensitization or food allergies were found. CONCLUSIONS: We found no evidence that maternal intervention with prebiotics protects against AD at 1 year of age in infants at risk of allergic diseases. Plain Language Summary: Atopic dermatitis is a skin condition that causes skin to become red, itchy and inflamed. It is also known as eczem

OBJECTIVE: Postprandial hyperglycemia is a major risk factor for type 2 diabetes and cardiovascular disease. Inhibition of α-amylase and α-glucosidase can attenuate postprandial glycemic response (PPGR). This study aimed to investigate the inhibitory effects of mulberry leaf and corn silk on these enzymes in vitro and their impact on postprandial glucose (PG) levels in prediabetic individuals using milk-based matrices. RESEARCH DESIGN AND METHODS: In vitro, enzyme inhibition was assessed using the DNS method (α-amylase) and pNPG method (α-glucosidase). A randomized crossover trial was conducted in 11 prediabetic individuals with four interventions: pure milk; lactose-hydrolyzed milk; lactose-hydrolyzed milk with mulberry leaf, corn silk, and resistant dextrin; and GOS milk with mulberry leaf and corn silk. PPGR was assessed by area under the glucose curve, 1 and 2 h PG, maximum PG, and 2 h glucose excursion. Paired Wilcoxon signed-rank tests were used for comparisons. RESULTS: Mulberry leaf and corn silk extracts inhibited both enzymes dose-dependently, with synergistic effects. No significant differences in PPGR indices were observed across interventions in the overall prediabetic individuals. However, in the overweight subgroup, the combination of GOS milk supplemented with mulberry leaf and corn silk significantly reduced 1 h PG (median difference [P25, P75]: -0.84 mmol/L [-1.05, -0.49]), maximum PG (-0.54 mmol/L [-0.75, -0.25]), and glucose excursion (-0.62 mmol/L [-0.75, -0.24]) compared to pure milk. CONCLUSIONS: Mulberry leaf and corn silk extracts inhibit α-amylase and α-glucosidase in vitro and may attenuate postprandial glucose excursions in overweight prediabetic individuals when delivered in a GOS milk matrix.

Background/Objectives: Ingestion of galactooligosaccharides (GOSs) or GOS mixtures has been purported to improve exercise-induced gastrointestinal (GI) distress and post-exercise recovery. However, the effects have not been explored in recreationally trained endurance athletes. This triple-blind randomised controlled trial, therefore, investigated whether 12 weeks of B-GOS® supplementation affects gastrointestinal comfort and psychological wellbeing in recreational athletes. Methods: Eighteen physically active individuals (12 males, 8 females, 44 ± 14 years, 1.7 ± 0.1 m and 73 ± 14 kg) volunteered for this study. Participants were assigned to independent groups in a placebo-controlled, triple-blind manner via stratified randomisation. A 20 min run at 80% VO2max was completed, with measures for GI distress and Competitive State Anxiety Inventory-2 questionnaire (CSAI-2) pre- and post-exercise. A 12-week supplementation period then ensued, where participants ingested either 3.65 g of B-GOS or an appearance-matched maltodextrin placebo. During this time, physical activity levels (IPAQ-7), general stress (REST-Q), mental wellbeing (WEMWBS), and sleep (core consensus sleep diary) were measured at regular time points. Results: There were no significant differences in VO2max (p = 0.437), GI discomfort (p = 0.227), or CSAI-2 (p = 0.739-0.954) from pre- to post-exercise at any time point or between conditions. Over the 12 weeks there were no significant differences between B-GOS and placebo in IPAQ-7 (p = 0.144-0.723), REST-Q (p = 0.282-0.954), WEMWBS (B-GOS pre = 51 ± 10, post = 53 ± 7; PLA pre = 51 ± 4, post 54; p = 0.862), or sleep (p = 0.065-0.992). The linear mixed model suggests that some may benefit on an individual level in terms of WEMWBS, general stress score, recovery-related scores, sleep, and sport-specific recovery score. Conclusions: There were no group benefits of B-GOS supplementation compared with placebo, although the individual variation may warrant further research in larger sample sizes and longer-duration studies.

Background and Objectives: The development of non-dairy probiotic products is a challenge for the food industry, while cereals, as probiotic carriers, provide the means to incorporate probiotics, prebiotics, and fiber into the human diet. The present study investigated the effects of Lactococcus cremoris spp. immobilized on oat flakes on blood and urine biomarkers in a randomized placebo-controlled single-blind clinical trial. Materials and Methods: Fifty-four eligible participants were randomized into a placebo or probiotic group that consumed 5 g of oat flakes daily for 12 weeks. Blood and urine samples were collected at the baseline, 6 weeks, and 12 weeks to assess the glycemic, lipemic, inflammatory, immunological, and antioxidant biomarkers, as well as the vitamin levels. Results: The intervention group exhibited a significant reduction in their hs-CRP levels (p = 0.002) and a trend toward decreased IL-6 levels (p = 0.035) at week 12 compared to the control group, suggesting a potential anti-inflammatory effect. Additionally, a significant reduction in insulin levels was observed in the probiotic group at week 6, with a clinical trend toward differentiation despite the absence of statistically significant differences between the groups. Furthermore, there were promising results regarding certain biomarkers, such as vitamin B12 and cortisol levels, in the probiotic group. Conclusions: The twelve-week consumption of Lactococcus cremoris spp. immobilized on oat flakes resulted in improvements in inflammatory, metabolic, and stress-related biomarkers. These results support the examined concept of non-dairy probiotic products, though further research is needed to confirm their efficacy and clarify their underlying mechanisms.

Galacto-oligosaccharides (GOS) are prebiotics that modulate gut microbiota and are implicated in the gut-brain axis (GBA), with preclinical models reporting effects on neurochemistry, brain function, and cognition. Here we report the results of a randomised, double-blind, placebo-controlled trial in 83 healthy females (17-25 years), who received GOS or placebo for 28 days. Assessments occurred at baseline, endline, and 28 days post-supplementation. The primary outcome was trait anxiety, secondary outcomes were brain-based levels of GABA and glutamate in the dorsolateral prefrontal cortex (dlPFC), anterior cingulate cortex, and inferior occipital gyrus (IOG) (measured with 1H-MRS), and gut microbiome composition. Tertiary outcomes included social anxiety, depression, emotion behaviour, reaction times, and nutritional intake. Analyses included intention-to-treat, per-protocol, and sensitivity approaches. Trait anxiety did not differ between groups at endline (p = 0.443), though trends favoured lower anxiety in the GOS group at follow-up (p = 0.069). GOS reduced GABA at trend significance in the IOG (p = 0.053) in the Intention to Treat (ITT) population and dlPFC (p = 0.088) in high-anxious participants, with effects persisting at follow-up. GOS transiently increased Bifidobacterium abundance (p = 0.001) but did not affect microbiome diversity. Tertiary outcomes showed no significant changes in social anxiety or depression but faster reaction rates in high-anxious participants for simple (p = 0.036) and choice tasks (p < 0.001). Nutritional intake was unaffected. While GOS supplementation did not significantly reduce trait anxiety, it produced neurochemical changes and transient modulations of the gut microbiome in Bifidobacterium abundance, indicating GOS-induced changes can be traced along the GBA.

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.

BACKGROUND: In this double-blind placebo-controlled randomised intervention we investigated the potential benefits of a prebiotic supplement on children's well-being in a home setting. The primary aim was to determine if this supplement could effectively reduce anxiety, improve mood, and enhance cognitive function, similar to findings in young adults. METHODS: Fifty-three healthy children, aged 6 to 14, participated in an 8-week trial. The trial consisted of three testing time points; day zero marked the baseline measurement (T1) followed by a 28-day supplement intervention period during which they consumed 5.5 g of the prebiotic galactooligosaccharides (GOS) daily under parental guidance. Endline measures (T2) were conducted on the last day of supplement consumption, with a final follow-up testing session (T3) on day 56. Primary outcomes were trait anxiety using a questionnaire and emotional behavior in a dot-probe task on responses to positive and negative images. Secondary outcomes encompassed depression levels, cognitive function tests, and dietary intake recorded in a 4-day food diary. Additionally, we explored whether parents' emotional behavior had an impact on children's responses. RESULTS: While our statistical analysis did not reveal significant effects of GOS, there were noteworthy trends. Trait anxiety levels decreased over time in both groups, with a more pronounced decrease in the GOS group (after intervention, p =.090; after follow-up, p =.031). The GOS group exhibited reduced negative emotional responses compared to the placebo group (p =.105), and post-trial depression levels decreased in the GOS group over time (p =.015). Although parental emotional responses correlated with various emotional outcomes in children, they did not influence the intervention effects. CONCLUSIONS: These findings suggest positive trends in line with our hypotheses, however further investigation with greater statistical power would be beneficial. TRIAL REGISTRATION: Retrospectively registered on https://clinicaltrials.gov/ [NCT06258135] on February 6, 2024. © 2025. The Author(s).

Prebiotics, traditionally linked to gut health, are increasingly recognized for their systemic benefits, influencing multiple organ systems through interactions with the gut microbiota. Compounds like inulin, fructooligosaccharides (FOS), and galactooligosaccharides (GOS) enhance short-chain fatty acid (SCFA) production, benefiting neurocognitive health, cardiovascular function, immune modulation, and skin integrity. Advances in biotechnology, including deep eutectic solvents (DES) for extraction and machine learning (ML) for personalized formulations, have expanded prebiotic applications. Integrating these innovations with "omics" technologies enables precise microbial modulation, fostering personalized nutrition and precision therapies. This review examines organ-specific effects of prebiotics, highlights findings from clinical trials, and explores biotechnological innovations that enhance prebiotic efficacy, laying the groundwork for future personalized therapeutic strategies. © 2025. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

AIM: To assess the efficacy of the combined administration of myo-inositol and zinc, a mineral involved in the insulin pathway, in paediatric obesity with insulin resistance on HOMA-IR, glucose-insulin metabolism, and lipid profile. MATERIALS AND METHODS: Double-blind, randomized, placebo-controlled study conducted in North Italy. Fifty-six patients (10-18 years, Tanner stage ≥3) with obesity and insulin resistance were randomized to myo-inositol (2000 mg), zinc gluconate (5 mg), and galactooligosaccharides (GOS) from plant-based origin (1000 mg) (TRT) or placebo (PLC) containing only GOS from plant-based origin (1000 mg). All patients received an isocaloric diet following the Mediterranean diet style. Data were collected at baseline (V0) and after 3 months (V1). The primary outcome was the insulin resistance index (HOMA-IR). RESULTS: Fifty out of 56 recruited subjects completed the study. TRT improved HDL cholesterol level compared to PLC (p = 0.05) but not insulin resistance. A stratified post hoc analysis was performed by sex, BMI, and subgroups of adherence to the Mediterranean diet. Subjects were divided for obesity grade, fasting insulin (p = 0.0137) and HOMA-IR (p = 0.0273) were lower in TRT than in PLC patients, with a greater effect on severe obesity. No adverse events were detected. CONCLUSION: Three months of supplementation with myo-inositol and zinc were beneficial on lipid profile and in managing obesity complications at least in subjects with severe phenotype. Thus, myo-inositol and zinc could be used as non-pharmacological agents. This work suggests a long-term study with a larger sample size to enrich the findings. © 2025 John Wiley & Sons Ltd.

BACKGROUND AND AIMS: Combining probiotics and prebiotics in synbiotics may present a synergistic approach to improve type 2 diabetes mellitus (T2DM); however, further evidence is required to establish the comparative efficacy of synbiotics versus probiotics. This study aimed to evaluate the effects of Bifidobacterium animalis subsp. lactis MN-Gup (MN-Gup) and a synbiotic mixture of MN-Gup and galactooligosaccharide (MN-Gup-GOS) on glycemic control in T2DM patients and explore possible mechanisms. METHODS: This randomized, double-blind, placebo-controlled clinical trial assigned 120 T2DM patients, to receive MN-Gup, MN-Gup-GOS, or placebo intervention for 12 weeks. The primary outcome was fasting blood glucose (FBG), with secondary outcomes including hemoglobin A1C (HbA1C), insulin, homeostatic model assessment of insulin resistance (HOMA-IR), inflammatory indicators, oxidative stress indicators, gastrointestinal hormones, gut microbiota, and bile acids (BAs). RESULTS: The median age of the 120 participants was 59 years (interquartile range: 55-62 years), with 40 being men. Compared to baseline, all three groups exhibited significant reductions in FBG. Additionally, the MN-Gup-GOS group demonstrated significant decreases in HbA1c, serum insulin, and HOMA-IR after intervention, whereas no such reductions were observed in the placebo and MN-Gup groups. Regarding the between-group comparisons, the MN-Gup-GOS intervention showed a significantly greater reduction in FBG compared to the placebo (least squares mean difference [95 % CI], -0.69 [-1.29, -0.10] mmol/L, P = 0.022) and MN-Gup (-0.59 [-1.17, -0.01], P = 0.047) group, but not for other indicators of glucose metabolism. Additionally, MN-Gup and MN-Gup-GOS intervention, especially the latter, significantly modified inflammation, oxidative stress, gut microbiota, serum BAs, and GLP-1 levels. Correlation analysis showed significant associations between changes in certain gut microbiota (Bifidobacterium) and BAs (deoxycholic acid and lithocholic acid) with glycemic indicators. CONCLUSIONS: The auxiliary effect of synbiotics MN-Gup-GOS on reducing FBG levels surpassed that of MN-Gup probiotics alone in T2DM patients,

Iron fortification compounds are of special interest to treat iron deficiency anemia, however, the dose-response effects of these fortificants on liver and renal functions have not been extensively reported in human subjects. The present study determines the effects of prebiotics and iron fortificants on liver function tests (LFTs) and renal function tests (RFTs) among women of reproductive age (WRA). A double-blind randomized controlled trial was performed for the duration of 90 days. A total of 75 iron-deficient women were selected and randomly divided into 5 groups (4 treatment groups and 1 control group). For this purpose, four different types of fortified wheat flour were prepared using two iron fortificants (NaFeEDTA and FeSO4) and two prebiotics (Inulin and Galacto oligosaccharides) were given to four treatment groups, while control groups were only given iron-fortified flour without the addition of prebiotics. Blood samples were collected every month to evaluate Liver Function Tests, including Alanine Transaminase (ALT), Aspartate Transaminase (AST), Alkaline Phosphatase (ALP), total bilirubin and Renal Function Tests, including serum urea and creatinine. Our results found that prebiotic and iron-fortified diets increased ALT, AST and total bilirubin levels among WRA. For AST, ALP and total bilirubin, our results found the highest increase in the treatment groups treated with prebiotics and iron fortificants at 963 mg/kg GOS + 15 ppm FeSO4. Moreover, the highest values of ALT and serum creatine were seen in groups treated with 963 mg/kg Inulin + 20 ppm NaFeEDTA, while maximum value for serum urea could be seen in the group given 963 mg/kg GOS + 30 ppm FeSO4. The study concluded that prebiotic and iron-fortified diets increased ALT, AST and total bilirubin levels among WRA.

Chronic stress is a critical predisposing factor for neuropsychiatric and metabolic disorders, profoundly affecting both physical health and behavioral performance. Stress induces gut microbiota dysbiosis and disrupts tryptophan metabolism, notably via the kynurenine (KYN) and 5-hydroxytryptamine (5-HT) pathways. Targeted modulation of the gut-brain axis through probiotics and prebiotics has therefore emerged as a promising approach for preventing and alleviating stress-related disorders, yet the efficacy of specific synbiotic combinations remains largely unexplored. This study investigated the effects of feline milk-derived Lactiplantibacillus plantarum MNN, galacto-oligosaccharides (GOS), and their combination on stress-related behavioral abnormalities and tryptophan metabolism in a mouse model of chronic social defeat stress (CSDS). Behavioral outcomes, gut microbial composition, cecal tryptophan metabolites, and molecular markers associated with serotonin (5-HT) synthesis and neuroplasticity were evaluated. Intervention with MNN, GOS, and MNN + GOS attenuated several CSDS-induced behavioral abnormalities and was associated with alterations in gut microbial composition and cecal tryptophan metabolism. In the SIT, the social interaction ratio increased from 0.307 ± 0.089 in CSDS mice to 0.778 ± 0.042 (p = 0.003), 0.826 ± 0.035 (p = 0.001), 1.445 ± 0.114 (p < 0.0001) in the MNN, GOS, and MNN + GOS groups, respectively. Cecal indoxyl sulfate levels decreased from 1.944 ± 0.286 in CSDS mice to 0.206 ± 0.022 (p < 0.0001), 0.231 ± 0.058 (p < 0.0001) and 0.275 ± 0.018 (p < 0.0001) in the MNN, GOS, and MNN + GOS groups, respectively. The interventions also increased intestinal and hippocampal 5-HT signals and partially restored the expression of tryptophan hydroxylases, brain-derived neurotrophic factor (BDNF), and postsynaptic density protein 95 (PSD95). These findings suggest that MNN, GOS, and MNN + GOS interventions may alleviate stress-related abnormalities, potentially through modulation of the microbiota-tryptophan-5-HT axis. However, the causal role of the gut microbiota and the efficacy of these interventions in cats require further investigation.

GH2 β-galactosidases are important industrial biocatalysts for lactose hydrolysis and prebiotic galacto-oligosaccharide (GOS) synthesis. Here, lacZ genes from Streptococcus thermophilus RB16 (StLacZ) and Lactobacillus delbrueckii subsp. bulgaricus NSP1 (LbLacZ) were cloned, sequenced and deposited in GenBank under accession numbers QB046854 and QB046855, respectively, and expressed in Escherichia coli BL21 (DE3). A comparative study was conducted to determine their catalytic activity. rLbLacZ was found to be a typical mesophilic enzyme, with the optimal pH of 6.5 and a temperature of 40 °C, and high apparent affinity for lactose (Km = 14.02 mM). Nevertheless, rStLacZ exhibited an alkaline optimum and high thermo-activity, with optimal catalytic rates at pH 9.0 and 55-60 °C, as well as a high turnover number (kcat = 128.82 s-1) and specific activity (63.5 U/mg) for lactose. Despite different functional substrate affinities and reaction rates, both enzymes proved to have similar overall catalytic efficiency. The enzymes required specific divalent metals and monovalent metal salts to achieve peak catalytic activity. rStLacZ catalyzes the rapid and early synthesis of fast-migrating transglycosylation intermediates, while rLbLacZ catalyzes the progressive formation of slower-migrating, more complex oligosaccharide fractions. These findings demonstrate the complementary industrial potential of rStLacZ for rapid bioprocesses at elevated temperatures and rLbLacZ for the gentle processing of dairy products.

The increasing demand for lactose-free dairy products, sustainable whey valorization and high-value prebiotic ingredients has intensified the need for efficient bacterial β-galactosidases capable of both lactose hydrolysis and galacto-oligosaccharide (GOS) synthesis. Although numerous bacterial β-galactosidases have been identified and recombinantly produced, information on their structural characteristics, catalytic properties, recombinant production efficiency and performance under application-relevant conditions remains scattered across individual studies, complicating their comparative evaluation for specific biotechnological applications. This review presents a comprehensive comparative overview of recombinant bacterial β-galactosidases with particular emphasis on their potential for lactose bioconversion and GOS production. Current knowledge on enzyme diversity, glycoside hydrolase families, structural organization, molecular determinants of catalysis, recombinant expression systems, purification strategies, hydrolytic activity and transglycosylation capacity is critically evaluated together with recent advances in protein engineering, computational enzyme design and commercially available β-galactosidase preparations. Comparative analysis of published studies indicates that catalytic efficiency alone is insufficient to assess the potential suitability of β-galactosidases for industrial applications. Enzyme performance is influenced by the combined effects of molecular architecture, catalytic properties, recombinant production efficiency, thermostability, product inhibition and transglycosylation capacity. However, direct comparisons remain limited by differences in experimental conditions and by the relatively small number of studies conducted in real dairy matrices or under industrially relevant process conditions. Accordingly, this review compares the major biochemical, molecular, recombinant production and process-related factors that should be considered when evaluating β-galactosidases for lactose hydrolysis, GOS production and large-scale bioprocessing. Particular attention is given to the current limitations in translating laboratory-scale enzyme c

Although high internal phase emulsions (HIPEs) have shown potential for probiotic encapsulation, most reported systems mainly provide structural protection and lack multifunctional regulation of gastrointestinal stresses. This study aimed to develop a multifunctional egg-derived HIPEs platform co-stabilized by egg yolk granules (EYGs) and salted egg whites (SEWs), integrating probiotic encapsulation, localized acid-buffering capacity provided by eggshell powder (ESP), and nutritional support to enhance the gastrointestinal delivery efficiency of probiotics. The optimal formulation for Lacticaseibacillus rhamnosus GG (LGG)-loaded HIPEs was identified by particle size and zeta potential measurements together with plate count, corresponding to the system containing 3% galactooligosaccharides (GOS) and 2% LGG. Under this formulation, LGG/GOS@HIPEs exhibited the superior storage stability and probiotic viability after 30 d at 4 °C. The in vitro simulated digestion results demonstrated a marked difference in probiotic survival among the tested groups. Following SGF treatment, LGG/GOS/ESP@HIPEs and LGG/GOS@HIPEs showed lower reductions (0.73 and 0.86 log CFU/mL) than free LGG (2.92 log CFU/mL). Crucially, LGG/GOS/ESP@HIPEs still maintained a viable count of 1.52 ± 0.20 × 106 CFU/mL after gastrointestinal digestion, corresponding to six orders of magnitude increase in survival relative to free probiotics, which were almost completely inactivated. To further evaluate gastrointestinal retention, ICG-labeled probiotics were tracked by In Vivo Imaging System. The system reduced gastrointestinal loss and enhanced intestinal retention of LGG in mice, with nearly 2-fold higher gastrointestinal fluorescence intensity than free probiotics at 15 h. These findings provide a food-grade HIPEs-based strategy for the enhancement of oral probiotic delivery efficiency.

High sucrose content and rapid staling limit the quality of fermented rice cakes (FRCs). This study evaluated the effects of substituting sucrose with xylooligosaccharides (XOS), fructooligosaccharides (FOS), and galactooligosaccharides (GOS) at different ratios (10%-40%) on the quality, starch retrogradation, and starch digestion properties of FRCs. Results showed that oligosaccharides delayed the pasting of rice flour and reduced its gelatinization enthalpy. Notably, a 20% substitution optimized batter gas retention, increasing specific volume and elasticity while reducing hardness. Moreover, gas chromatography-ion mobility spectrometry revealed that XOS, FOS, and GOS promoted the generation of fermentation metabolites, enriching the flavor profile with fruity ester aromas. During storage, oligosaccharides restricted water migration from bound to free states and retarded both short-range starch rearrangement and long-range crystal growth, effectively delaying hardening. Furthermore, XOS increased resistant starch content in FRCs by approximately 57.8%. These findings suggest that moderate replacement of sucrose with oligosaccharides is a promising strategy for delaying staling and improving the overall quality of FRCs.

Lupin protein isolate (LPI) is limited in food use by poor solubility and weak interfacial behavior. This study tested whether the extraction route conditions LPI for subsequent glycation. LPI was recovered by microwave-assisted extraction, applied only during extraction, and conjugated with galacto-oligosaccharides (GOS) by wet heating at 90 °C for 15 min. Spectroscopic, electrophoretic, thermal and microscopic analyses confirmed GOS attachment and partial unfolding. The conjugate from microwave-extracted LPI reached the highest apparent degree of grafting (22%), reduced particle size from 1775.1 to 89.7 nm and surface hydrophobicity from 443.3 to 62.5 arbitrary units, and lowered β-sheet content. Foaming (64%/36%), emulsifying activity (74.2 m2/g) and DPPH scavenging (79.6%) improved accordingly. In fecal fermentation it retained more acetate at 24 h (420.8 versus 212.1 μmol/L for the control) and gave the highest gas volume (13.9 mL) and microbial evenness. Extraction history therefore governs conjugate functionality. © 2026 The Authors.

Prebiotics are nondigestible food components that selectively stimulate beneficial gut microbiota and have garnered significant attention in food science and biotechnology because of their health-promoting properties. Among them, fructooligosaccharides and galactooligosaccharides are especially important for their positive effects on human health. Functional foods with enhanced nutritional and medicinal value have been made possible by the recent use of prebiotics in food fermentation and bioprocessing. This review summarizes recent developments in prebiotic production and application, with emphasis on biotechnological approaches such as microbial fermentation, enzymatic synthesis, and strain improvement techniques, based on literature retrieved from PubMed, Scopus, and Web of Science up to July 2025. Advances in fermentation technologies, including solid-state fermentation and co-culture systems, are also discussed for optimizing production processes. In addition, the review highlights the health-promoting effects of prebiotics, including modulation of gut microbiota, immunological regulation, and involvement in the gut-brain axis. Lastly, we highlighted current challenges and prospects, emphasizing their application to transform fermented foods and bioprocessing techniques.

A novel ultrasound-assisted synbiotic microencapsulation system was developed using gum arabic, reconstituted skim milk, and prebiotic oligosaccharides (fructooligosaccharides, xylooligosaccharides, and galactooligosaccharides) for the delivery of Clostridium butyricum. Ultrasound significantly improved viable-cell recovery, reaching 93.17 % in the fructooligosaccharide formulation (p < 0.05). Structural analyses indicated that oligosaccharide type and ultrasound treatment affected particle morphology, water mobility, protein secondary structure, and the solvent-extractable fractions related to intermolecular interactions. Ultrasound treatment improved the environmental stress tolerance of the freeze-dried formulations, increasing 60-day storage survival by 10.4 %, 6.9 %, and 16.6 % in the control, XOS, and GOS formulations, respectively, and significantly reducing viability loss during separate gastric, intestinal, and bile-salt tolerance tests (p < 0.05). Among the tested oligosaccharides, the XOS-containing formulations showed the most balanced overall performance. MC-XOS showed the highest survival after heat treatment (51.3 %) and retained a survival rate of 27.5% after 3 h of separate exposure to simulated gastric fluid, whereas MC-XOS-US maintained the highest survival after 60 days of storage at 4 °C (60.87 %) (p < 0.05). These findings provide an effective strategy for developing stable delivery systems for anaerobic probiotics.

Synbiotics exert various physiological effects by complementing the physiological functions of both probiotics and prebiotics. Chronic constipation is a prevalent global health concern, and despite several clinical studies utilising synbiotics, definitive conclusions regarding its efficacy remain elusive. This study aimed to investigate the effect of a synbiotic combination of Lacticaseibacillus paracasei strain Shirota (LcS: formerly known as Lactobacillus casei strain Shirota) and galactooligosaccharides (GOS), both of which independently improve defecation symptoms, on defecation frequency in constipation-prone healthy Japanese adults. This study employed a placebo-controlled, randomised, double-blind, parallel-group comparison design over a 4-week consecutive intake period. The primary objective was to evaluate the efficacy of the synbiotic intervention on bowel movement frequency in healthy adults prone to constipation. The synbiotic group exhibited a significantly increased defecation frequency at week 2 ( P = 0.048) but not at week 4, whereas the number of defecation days showed only a trend at week 2 ( P = 0.058). This represents a key limitation of this study. A significant positive correlation was observed between faecal LcS counts and defecation parameters in the synbiotic group, with no equivalent correlation observed for endogenous Lactobacillaceae in either group. This suggests a strain-specific effect of LcS beyond a general class effect of Lactobacillaceae. Additionally, increases in faecal Bifidobacterium counts and a decrease in isobutyric and isovaleric acid levels were observed along with improvements in defecation-related symptoms. These findings suggest that the combination of LcS and GOS may contribute to improvements in constipation-related outcomes in healthy individuals prone to constipation. The study was registered by the University Hospital Medical Information Network: registration number: UMIN000039236.

BACKGROUND: Milk oligosaccharides (MOs) play a crucial role in the development of the epithelial barrier function in newborn mammals. OBJECTIVES: This study aims to explore the effects of 3 synthetic MOs-galactooligosaccharides (GOS), 2'-fucosyllactose (2'-FL), and 3'-sialyllactose (3'-SL)-on the growth performance and intestinal health of recently weaned piglets. METHODS: Forty Duroc × Landrace × Yorkshire piglets (7.10 ± 0.25 kg) weaned at 21 d of age were randomly divided into 4 treatments and fed a basal diet (BD) or BD supplemented with 0.5% different MOs (GOS, 2'-FL, and 3'-SL). RESULTS: 2'-FL and 3'-SL significantly increased the growth performance of piglets as shown by decreases in the ratio of feed-to-gain and diarrhea ratio (P < 0. 05), whereas GOS showed limited effects on these indices. Compared with dietary GOS, the 2'-FL and 3'-SL supplementation induced a stronger digestibility of nutrients such as dry matter, crude protein, and crude fat, and also enhanced the activities of duodenal alkaline phosphatase and maltase as well as jejunal lactase (P < 0.05). Also, 2'-FL and 3'-SL not only increase the intestinal villus-to-crypt ratio but also elevate the abundance of zonula occludens-1 and secretory immunoglobulin A in the jejunal epithelium (P < 0.05). In addition, the abundance of total bacteria, Lactobacillus, and Bifidobacterium was numerically higher in the digesta of weaned pigs in the 2'-FL and 3'-SL groups than in those in the control group. Importantly, 2'-FL and 3'-SL selectively increased the ratio of CD4+/CD8+ cells in mesenteric lymph nodes (MLNs) but decreased the abundance of T helper 17 cells in MLNs and the content of interleukin (IL)-6 in serum. In addition, 2'-FL and 3'-SL supplementation increased the expression levels of SLC2A2 and SLC27A4 but significantly decreased the expression levels of IL-17A, IL-1β, and RELA in the jejunum (P < 0.05). CONCLUSIONS: These results suggested that MOs show a significant beneficial effect on weaned pigs in a structure-dependent manner, because 2'-FL and 3'-SL were more effective than GOS, partly by improving nutrient absorption and intestinal barrier function.

Galacto-oligosaccharides (GOS) are widely recognized as prebiotic ingredients, whereas their technological role in stabilizing lipid-based systems remains poorly understood. In this study, GOS were synthesized by enzymatic transgalactosylation of lactose and enriched by ethanol precipitation. Carbohydrate composition, glass transition temperature (Tg), and their influence on the formation of primary lipid oxidation were evaluated using freeze-dried lecithin liposomes as a model membrane system. Ethanol precipitation increased the GOS content from 22.8% to 50.7% (w/w) while reducing the proportion of monosaccharides from 40.0% to 15.5% (w/w). Tg was strongly influenced by carbohydrate composition and relative humidity (RH), with GOS-enriched preparations exhibiting higher Tg values than the crude reaction mixture. The formation of primary lipid oxidation products, assessed by UV spectroscopy through conjugated diene formation at 234 nm, was significantly lower in liposomes freeze-dried with GOS-rich preparations or sucrose than in phosphate-buffered saline (PBS) controls, particularly under storage conditions in which the carbohydrate matrices remained in the glassy state. At higher RH, transition to the rubbery state was associated with increased conjugated diene formation. These findings suggest that the protective effect of GOS-rich formulations is associated with the physicochemical properties of the carbohydrate matrix and is consistent with the proposed role of the glassy state in limiting molecular mobility, rather than direct antioxidant activity. Beyond their established prebiotic properties, GOS-rich preparations may also contribute to the stability of dehydrated lipid-containing systems, supporting their use as multifunctional food ingredients.

Antibiotics are widely used to prevent and treat early life gut infections, but their aberrant use has raised global concerns about antibiotic resistance. As breastfed infants have fewer intestinal infections than formula-fed infants, this stimulated scientific interest in the anti-pathogenic properties of breastmilk, specifically human milk oligosaccharides (HMOs). HMOs are non-digestible sugars that support infant gut microbiome development, immune maturation, and protection against pathogens. However, to determine how specific HMO structures differentially affect microbial growth and pathogen susceptibility investigating individual HMOs is essential. This study therefore evaluates the effects of single-structure HMOs and galacto-oligosaccharides (GOS) on the growth of key early-life gut pathogens, Escherichia coli, Salmonella enterica, Yersinia enterocolitica, and Clostridium perfringens. The strongest pathogen inhibitory effects of HMOs were further assessed by colony forming unit enumeration and tested across variable media to confirm oligosaccharide-specific efficacy. Oligosaccharide screening revealed clear differences in bacterial growth, with S. enterica most strongly inhibited and C. perfringens exhibiting growth stimulation. Among the tested oligosaccharides, 3'-galactosyllactose and GOS showed the most pronounced growth-inhibitory effects against Escherichia coli, Salmonella enterica, Yersinia enterocolitica. Salt concentration and nutrient richness further modulated oligosaccharide activity, highlighting the multifactorial nature of their antibacterial potential. Overall, our findings show that oligosaccharides display structure-specific and environment-dependent effects on bacterial growth, indicating that their antimicrobial activity is shaped by oligosaccharide structure, bacterial species, and growth environment.

The global use of nutraceuticals and dietary supplements-including vitamins, minerals, probiotics, and botanical extracts-has increased substantially, driven largely by consumer perceptions of safety and health benefits. However, their potential to cause hypersensitivity reactions (HSRs) is underrecognized. This review synthesizes published case reports, case series, systematic reviews, and pharmacovigilance data to characterize HSRs linked to these products, identify risk factors, and examine diagnostic and regulatory challenges. We classified reactions as immediate (< 1-6 h after administration), non-immediate (> 1-6 h after administration to several weeks). Immediate HSRs, including anaphylaxis, have been reported after vitamins B complex, short-chain galacto-oligosaccharides (GOS), whole-food supplements such as spirulina, natto (poly[γ-glutamic acid]) and royal jelly, and botanical/plant derived products; diagnosis commonly used skin-prick and intradermal testing and basophil activation tests. Non-immediate HSRs-ranging from maculopapular eruptions to severe cutaneous adverse reactions-have been associated with cobalamin, chromium picolinate, alpha-lipoic acid, diindolylmethane (DIM), glucosamine/chondroitin, and euglena; diagnosis often relies on patch testing, lymphocyte transformation assays, and cautious drug provocation when indicated. These products can also cause allergic contact dermatitis, and occupational sensitization (e.g., rhinitis/asthma) has been reported in workers exposed to psyllium and Aspergillus oryzae-derived lactase. Diagnostic accuracy is limited by nonstandardized reagents, variable test concentrations, complex product formulations, and cross-reactivity (notably within botanical families such as Asteraceae and among plant proteins/legumes), complicating interpretation and increasing risk for atopic individuals. Evidence remains dominated by case-based reports and passive surveillance. To reduce risk, we recommend heightened clinician awareness, development of standardized diagnostic reagents and protocols, mandatory adverse-event reporting, clearer ingredient labeling, and premarket allergenicity assessment.

This study investigated the development of ultrasound-pretreated lupin protein isolate-galactooligosaccharide (LPI-GOS) conjugates by wet-heat Maillard glycation and evaluated their structural, techno-functional, bioactive, and in vitro fecal fermentation responses. The degree of grafting increased significantly from 29.8% in the unheated LPI-GOS mixture to 56.2% in the wet-heat conjugate and 64.1% in the ultrasound-pretreated wet-heat conjugate. Conjugation induced marked structural changes, including a reduction in particle size from 2738.2 nm in native LPI to 204.9 nm in ULPG-WH, a moderate shift in zeta potential from -25.5 mV in native LPI to -37.3, -32.2, and - 30.7 mV in LPG-M, LPG-WH, and ULPG-WH, respectively, and a decrease in surface hydrophobicity from 559.3 in native LPI to 158.6 in ULPG-WH. These structural modifications were associated with clear improvements in functional properties. Protein solubility increased from 50.43% in native LPI to 78.4% in the ultrasound-pretreated wet-heat conjugate, foaming capacity from 34.1% to 92.1%, and water-holding capacity from 6.9 to 11.4 g/g. The conjugates also showed enhanced biological functionality, including improved antioxidant activity and increased ACE-inhibitory activity, with the strongest effects generally observed in ultrasound-pretreated conjugates. During short-term in vitro fecal fermentation, the LPI-GOS systems influenced early microbial fermentation responses by favoring saccharolytic taxa, including Bifidobacterium and Faecalibacterium, and by increasing SCFA-associated fermentation patterns relative to the negative control. Because fermentation was limited to 24 h, these results should be interpreted as short-term substrate-driven responses rather than stable long-term gut microbiota shifts. Overall, ultrasound pretreatment combined with Maillard glycation effectively modified LPI structure and improved its functional and short-term fermentation-related properties, supporting its potential use in the development of multifunctional food ingredients.

Milk oligosaccharides (MO) support intestinal, microbial, and immune development in young pigs. However, modern production practices wean pigs at an early age, removing them from their source of MO prior to intestinal and immune maturation. The purpose of this study was to investigate the effects of dietary supplementation of galacto-oligosaccharides (GOS) and 2'-fucosyllactose (FL) on the jejunal mucosa-associated microbiota, intestinal immune signaling, morphology, and growth performance of nursery pigs. Forty-eight pigs (6.8 ± 0.2 kg body weight) weaned at 3-weeks-of-age were allotted into six dietary treatments, using a randomized complete block design, with sex and initial body weight as blocks. Dietary treatments were (1) basal diet; (2) basal diet, with supplemental GOS at 1.5% of the diet; (3) basal diet, with supplemental FL at 0.2% of the diet; (4) basal diet, with GOS and FL at 1.5 and 0.2% of the diet, respectively; (5) basal diet, with GOS at 2.3% of the diet; and (6) basal diet, with FL at 0.3% of the diet. These MO were provided alone or in combination at levels mimicking intake at the end of the suckling period, and at 1.5-fold higher, to observe potential dose-dependent responses. Pigs were fed for 21 days in two phases. On d 21, pigs were euthanized for sampling of jejunal mucosa and jejunal tissue. Data were analyzed using the PROC MIXED of SAS 9.4 and contrasts were used to determine the effects of GOS, FL, and their combination (interaction), in addition to the linear effects of increasing dietary GOS or FL. Increasing levels of GOS and FL linearly decreased (P < 0.05) Shannon and Simpson alpha diversity of the jejunal mucosa-associated microbiota. Supplementation with FL increased (P < 0.05) the absolute abundance of Helicobacter in the jejunal mucosa-associated microbiota, although no dose response was observed. Increasing levels of GOS and FL tended to linearly decrease (P = 0.051 and 0.076, respectively) the gene expression of TLR4. Increasing levels of GOS tended to increase (P = 0.054) and increasing supplementation of FL increased (P < 0.05) the number of Ki-67 proliferative cells in the crypt of the jejunum. Increasing levels of GOS

Cold-active β-galactosidase has significant application value in the enzymatic synthesis of galacto-oligosaccharides (GOS); however, its low transglycosylation efficiency and poor selectivity toward high-degree polymerization (DP) products remain major challenges. In this study, a cold-active β-galactosidase (K3-β-gal) from marine Bacillus sp. K-3 was cloned and expressed. Four mutants (R252M, F261W, Y281R, and F355K) were constructed via semi-rational design, among which R252M exhibited markedly enhanced transglycosylation activity. Compared with the wild-type enzyme, R252M showed no change in optimum temperature (20 °C) or optimum pH (6.0), and retained good stability under low-temperature and weakly acidic conditions. The total GOS yield increased from 21.68% to 38.12% (a 75.83% improvement), and the DP4/DP3 ratio increased from 0.95 to 2.6, while the hydrolytic activity (kcat/km) decreased by approximately 44%, indicating a shift in the catalytic equilibrium from hydrolysis toward transglycosylation. Molecular dynamics simulations revealed that R252M remodels the hydrogen-bond network in the active pocket, enhances protein rigidity, and reduces dynamic perturbations around the active site, which may contribute to substrate binding and transglycosylation efficiency.", This mutant holds promising potential for the green synthesis of high-DP GOS. © 2026. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.

In recent years, nutrition and health have become topics of widespread societal concern, and galactooligosaccharides (GOS) have garnered significant interest as prebiotics. Composed of one to nine galactose units terminated by a glucose molecule, GOS exhibit remarkable thermostability, high water solubility, strong humectancy, and low caloric value. They provide various physiological benefits, including prebiotic activity, gut health maintenance, antibacterial, antiadhesive, and anti-inflammatory effects, as well as systemic regulation of blood sugar, lipids, immune homeostasis, and metabolic diseases. These properties support broad applications in the food industry, breeding field, material science, and medical science. Enzymatic production using β-galactosidases is the primary industrial method for GOS synthesis. This review provides an overview of recent advances in β-galactosidase biochemistry, crystal structures, catalytic mechanisms, molecular modification, immobilization, and heterologous expression, along with GOS physiological functions, production, applications, and whey utilization. Future opportunities for GOS research and development are also briefly discussed.

Polyphenols from Saskatoon berry are promising functional ingredients, yet their value depends on stability during processing, digestion, and transepithelial transfer. This study evaluated freeze-dried inulin (INU) and inulin/galactooligosaccharide (INU/GOS) matrices as carriers for purified Saskatoon berry phenolics. Powders containing 5-25% GOS were characterized by SEM, simulated gastrointestinal digestion, Caco-2 transport, antioxidant and anti-inflammatory assays, prebiotic testing, and cytocompatibility assessment. Initial phenolic retention remained high (88.5-92.7%). Digestion markedly reshaped the profile: apparent total polyphenol bioaccessibility reached 106.2-122.6%, anthocyanins were the least stable fraction, and phenolic acids together with selected flavonols dominated the post-digestive pool. Transepithelial transfer was selective; INU90:GOS10 showed the most favorable overall transfer profile, whereas INU80:GOS20 yielded the highest absolute phenolic content after transport. GOS-containing systems improved selected antioxidant, anti-inflammatory, and prebiotic responses, while pure inulin showed the highest cytocompatibility. Overall, carrier composition modulated the post-digestive functionality of Saskatoon berry polyphenols.

INTRODUCTION: Cannabis sativa L. (cannabis) is valued for its flowers, which are rich in bioactive compounds that impart medicinal and sensory properties. A common-practice during plant cultivation is the supplementation of fertilisers with biostimulants, which enhance growth and yield. Previous studies on tomato, strawberry, capsicum, and cannabis (grown for seed) have demonstrated that biostimulants containing fish hydrolysate (FH), Aloe vera (L.) Burm.f. extract, molasses, or triacontanol can improve yield, quality, and functional food value. Additionally, galactooligosaccharides (GoS) are proposed as a novel biostimulant that selectively stimulates beneficial microbes, akin to a plant prebiotic. METHODS: Two biostimulant complexes were compared by randomised controlled trial (RCT), within an environmentally controlled hydroponic growth system, for their effects on cannabis growth, yield, and flower quality. The two complexes tested were: BC1 comprising molasses, A. vera extract, and FH; and BC2 comprising GoS, A. vera extract, and triacontanol. RESULTS AND DISCUSSION: Both treatments improved yield (BC1: 1.17-fold, p = 0.097; BC2: 2.22-fold, p = 0.003), with BC2 also increasing flower size (1.28-fold, p = 0.034). Image analyses indicated that neither treatment substantially impacted flower colour. Near infrared indicated that both treatments increased primary amines and methyl containing hydrocarbons, and that BC2 also increased aromatic hydrocarbons. Volatile analysis indicated that BC1 increased a-terpinolene, borneol, terpineol, and valencene, whilst BC2 increased a-humulene, a-phellandrene, α-terpinene, β-caryophyllene, guaiol, limonene, ocimene, and total terpene content. These terpene changes suggest possible increases to flower relaxing effects (BC1), or anti-inflammatory effects (BC2). Odour prediction identified possible shifts to flower odour profiles, which may be associated with enhanced customer perceptions of quality and value. Overall, this study demonstrates the potential for biostimulants to enhance cannabis yield, phytochemical composition, and associated value.

The mucin barrier is essential for postnatal gut homeostasis, but whether galacto-oligosaccharides (GOS) regulate mucin glycosylation remains unclear. Here, we investigated the effects of early-life GOS supplementation on mucin O-glycans and intestinal barrier development in piglets using UPLC, MALDI-TOF-MS, immunofluorescence, lectin staining, and histomorphological analyses. GOS supplementation significantly increased jejunal villus width, mucosal thickness, goblet cell numbers, and MUC2 expression during early life. Glycomic profiling revealed selective remodeling of mucin O-glycans, characterized by increased core 1/3 and α2,3-sialylated glycans and reduced core 2/4 and α2,6-sialylated structures. These alterations were accompanied by upregulation of C1GALT1, GALNTs, and ST3GALs, and downregulation of ST6GALNACs and neuraminidases. Lectin staining further confirmed enhanced α2,3-sialylation and reduced α2,6-sialylation. Collectively, these results demonstrate that dietary GOS promotes intestinal barrier development and selectively modulates the composition and sialylation of mucin O-glycans, providing novel mechanistic insight into how prebiotics support gut health in early life.

Oligosaccharide prebiotics, such as inulin, fructooligosaccharides (FOS), and galactooligosaccharides (GOS), have demonstrated significant effects on gut microbiota and host health across in vitro, animal, and clinical studies. These studies consistently report an increase in beneficial bacteria, particularly Bifidobacterium and Lactobacillus, leading to higher production of short-chain fatty acids (SCFAs) such as acetate, propionate, and butyrate. These metabolic changes are linked to improved integrity of the epithelial barrier, reduced inflammatory signaling, modulation of immune responses, and enhanced metabolic balance. Biotechnological production methods, including enzymatic synthesis, microbial fermentation, and controlled depolymerization of plant polysaccharides, allow for precise control over the degree of polymerization and the types of glycosidic linkages. This control directly affects the fermentability, microbial selectivity, and functional effectiveness of the prebiotics. When incorporated into functional food systems, oligosaccharide prebiotics can enhance physicochemical properties such as texture, sweetness, and stability, all while maintaining their biological performance. Advanced delivery technologies, such as nano- and microencapsulation, improve thermal stability, resistance to gastrointestinal degradation, and targeted colon-specific release. Additionally, synbiotic formulations can further enhance the effectiveness of these prebiotics by promoting microbial colonization and sustained availability of SCFAs. Therapeutic benefits have been observed across various models of gastrointestinal health, metabolism, immune responses, and the gut-brain axis. These benefits involve mechanisms such as GPCR activation, histone deacetylase inhibition, and cytokine regulation. However, several challenges remain, including dose-dependent gastrointestinal intolerance, variability in individual microbiomes, degradation during processing, regulatory hurdles, and high costs of downstream processing. Overall, these findings highlight oligosaccharide prebiotics as versatile and scalable biotechnological ingredients, emphasizing the need for standardized produc

Galactooligosaccharides (GOS) are one of the internationally recognized prebiotic products, which have become a hot research focus in the field of biofoods because of their strong prebiotic, sugar substitution and inflammation alleviation functions. β-galactosidase (Bgal) of different microorganisms is utilized industrially in order to achieve the biosynthesis of GOS. Although the biosynthesis of GOS has been supported by certain technologies, there is still room for further improvement of its synthetic yield. This paper mainly introduces the function and biosynthesis of GOS and its research progress in recent years to enhance the yield of biosynthesis. This paper also combines the research progress in related fields in recent years, based on the basic theories of molecular biology and bioinformatics, discusses the research progress of green, innovative approaches including enzyme engineering, enzyme immobilization, surface display, and microbial fermentation on the synthesis of GOS.

As a major milk allergen with limited functionality, β-lactoglobulin (β-Lg) requires effective modification strategies to enhance its applicability. This study investigated the synergistic effects of pH-shifting pretreatment and glycation on the structure, functional properties, and allergenic potential of β-Lg. The protein was first treated under strongly acidic or basic conditions to induce conformational rearrangements, followed by glycation with galacto-oligosaccharides (GOS) or xylo-oligosaccharides (XOS). Pretreatment under acidic conditions enhanced the protein structural flexibility, exposing more glycation sites, especially for the linearly structured XOS. Acidic pH-shifting with XOS glycation significantly improved the emulsifying and foaming properties of β-Lg, while reducing its IgE-binding capacity. In comparison, basic pH-shifting plus glycation resulted in the greatest reduction in protein allergenicity through a synergistic masking effect, albeit with compromised functionality due to aggregation. Overall, the combined modulation of structure and conjugation provides an effective strategy to tailor the functionality and allergenicity of milk proteins.

As emerging environmental contaminants, micro- and nanoplastics (MNPs) raise neurotoxic concerns. However, the mechanisms underlying their induction of emotional disorders remain poorly understood. In particular, intervention-oriented and functionally validated strategies for prevention and control are still lacking. To address this, we conducted a 28-day repeated oral exposure study in mice using a gradient of polystyrene nanoplastics (PS-NPs; 2, 10, and 50 mg/kg/day) spanning environmentally relevant doses, systematically assessing impacts along the microbiota-gut-brain axis. Our results showed that PS-NP exposure induced dose-dependent anxiety- and depression-like behaviors, with fluorescence tracing revealing predominant gut accumulation and limited brain distribution. Mechanistically, exposure induced gut microbiota dysbiosis, intestinal barrier impairment, and lipopolysaccharide translocation, ultimately leading to systemic inflammation and neuroinflammation. Serum biochemical analysis showed that gut dysbiosis initiates host tryptophan metabolism toward the kynurenine pathway by triggering inflammation and subsequent indoleamine 2,3-dioxygenase 1(IDO1) activation. Consistently, hippocampal transcriptomic and biochemical analyses confirmed decreased 5-HT levels and suppression of the downstream 5-HT1A-cAMP-PKA-CREB-BDNF neurotrophic cascade, demonstrating comprehensive serotonergic disruption from substrate depletion to receptor dysfunction. Notably, the prebiotic galacto-oligosaccharides (GOS) restored intestinal homeostasis and ameliorated these anxiety- and depression-like behavioral and metabolic deficits, exerting protective effects by counteracting the above pathway. Our work delivers a from-mechanism-to-solution understanding: it deciphers how microplastics disrupt the gut-brain axis to cause neurotoxicity and identifies GOS maintenance of intestinal health as a key mitigative strategy against plastic pollution risks. Therefore, maintaining intestinal health, particularly through dietary GOS, represents a viable strategy to mitigate the neurotoxicity induced by plastic pollution.

Background: Gut microbiome-derived metabolites, particularly short-chain fatty acids (SCFA) and tryptophan derivatives, are central mediators of the gut-brain axis. This ex vivo study assessed how nutritional interventions impact such metabolites during early life, a critical period for neurodevelopment. Methods: The effects of galacto-oligosaccharides (GOS), nutrient blends (vitamins, minerals and amino acids) and their combinations were evaluated in the gut microbiomes of infants (2-4 months, n = 6) and young children (2-3 years old, n = 6) using the ex vivo SIFR® technology. Results: Baseline microbiome composition was age-dependent, with infants displaying lower α-diversity and greater interpersonal variability. After ex vivo incubation, nutrient blends increased the propionate/butyrate ratio and branched-chain fatty acids in young children and elevated several B-vitamins and amino acid-derived metabolites, including indole-3-carboxaldehyde, imidazoleacetic acid and pipecolinic acid. Combining nutrient blends with GOS exhibited potential synergistic effects on propionate (infants) and 2-hydroxyisocaproic acid (HICA, both age groups). GOS strongly stimulated Bifidobacteriaceae and increased metabolites linked to bifidobacterial metabolism like acetate, HICA, N-acetylated amino acids, aromatic lactic acids and acetylagmatine; in young children, butyrate and γ-aminobutyric acid (GABA) also increased. Conclusions: Combinations of GOS with nutrient blends impacted microbiome-derived metabolites associated with the gut-brain axis, with potential synergistic increases of metabolites with emerging roles in neurodevelopment, including GABA, acetylagmatine and HICA. Despite shared bifidogenic effects, differences between age groups indicate that microbiome maturity may influence responses to nutritional intervention. Future clinical studies are needed to determine whether these metabolite changes translate into neurodevelopmental benefits in vivo.