Structural elucidation of mulberry leaf oligosaccharide and its selective promotion of gut microbiota to alleviate type 2 diabetes mellitus

Two oligosaccharide fractions(MLO 2-1 and 2-2)were purified from enzymatic hydrolysate of mulberry leaf polysaccharide.The results of simulated digestion showed that MLO 2-2 was a digestible oligosaccharide,which could be degraded by human digestive juice;while MLO 2-1 possessed the non-digestible property in the upper gastrointestinal tract and performed the function by regulating the gut microbiota.Hence,MLO 2-1 was selected for the further analysis.The structure of MLO 2-1 was elucidated as follow:α-T-Glcp-(1→3)-β-Glcp-(1→5)-α-Araf-(1→5)-α-Araf-1→5)-α-Araf-(1→3)-α-(2-OAc)-Glcp-1.The in vitro fecal fermentation results showed that MLO 2-1 could modulate the composition of gut microbiota.Meanwhile,MLO 2-1 was effectively metabolized by fecal bacteria to produce lactate and short chain fatty acids,especially acetate and butyrate.The specific metabolic pathways of MLO 2-1 by gut microbiota were further illuminated.Gut microbiota analysis revealed that MLO 2-1 selectively promoted the growth of Ligilactobacillus murinus,a commensal bacterium presented a reduced level in T2DM mice.Animal experiments indicated that MLO 2-1 and L.murinus exhibited hypoglycemic activities.These results demonstrated that MLO 2-1 might alleviate T2DM by selectively accelerating the proliferation of L.murinus.

Physiological and Transcriptome Analysis Illuminates the Molecular Mechanisms of the Drought Resistance Improved by Alginate Oligosaccharides in Triticum aestivum L.

Alginate oligosaccharides(AOS)enhance drought resistance in wheat(Triticum aestivum L.),but the definite mechanisms remain largely unknown.The physiological and transcriptome responses of wheat seedlings treated with AOS were analyzed under drought stress simulated with polyethylene glycol-6000.The results showed that AOS promoted the growth of wheat seedlings and reduced oxidative damage by improving peroxidase and superoxide dismutase activities under drought stress.A total of 10,064 and 15,208 differentially expressed unigenes(DEGs)obtained from the AOS treatment and control samples at 24 and 72 h after dehydration,respectively,were mainly enriched in the biosynthesis of secondary metabolites(phenylpropanoid biosynthesis,flavonoid biosynthesis),carbohydrate metabolism(starch and sucrose metabolism,carbon fixation in photosynthetic organisms),lipid metabolism(fatty acid elongation,biosynthesis of unsaturated fatty acids,alpha-linolenic acid metabolism,cutin,suberine and wax biosynthesis),and signaling transduction pathways.The up-regulated genes were related to,for example,chlorophyll a-b binding protein,amylosynthease,phosphotransferase,peroxidase,phenylalanine ammonia lyase,flavone synthase,glutathione synthetase.Signaling molecules(including MAPK,plant hormones,H_(2)O_(2) and calcium)and transcription factors(mainly including NAC,MYB,MYB-related,WRKY,bZIP family members)were involved in the AOS-induced wheat drought resistance.The results obtained in this study help underpin the mechanisms of wheat drought resistance improved by AOS,and provides a theoretical basis for the application of AOS as an environmentally sustainable biological method to improve drought resistance in agriculture.

Synergistic effects of alginate oligosaccharide and cyanidin-3-O-glucoside on the amelioration of intestinal barrier function in mice

Emerging evidence shows that dietary oligosaccharides are important prebiotics that can improve intestinal flora,while dietary polyphenols can act directly on intestinal cells.However,information about their synergistic effects on gut health is still limited.In this study,alginate oligosaccharide(AOS)and cyanidin-3-O-glucoside(C3G)were selected as a common marine plant oligosaccharide and terrestrial plant polyphenol,respectively,to study their effects on intestinal health.The results show that,in comparison to their individual applications,the combination of AOS and C3G(mass ratio,3:1)displayed a stronger ability to up-regulate the expression of tight junction proteins,while enhanced intestinal epithelial barrier was also observed and levels of mucin-2 andβ-defensins were simultaneously increased in the intestinal mucus.Interestingly,the secretion of immunoglobulin A and immune-related cytokines were approximately doubled by the AOS+C3G mixture.In addition,the AOS+C3G mixture was found to be more conducive to the positive transformation of intestinal flora,which stimulated the growth of beneficial bacteria Akkermansia,Lachnospiraceae and Feacalibaculum while inhibiting the growth of harmful bacteria Helicobacter and Turicibacter.The data generated herein thus suggests that dietary oligosaccharides and dietary polyphenols may be more beneficial to intestinal health when applied in combination than their individual effects alone.

Mannan oligosaccharides alleviate oxidative injury in the head kidney and spleen in grass carp(Ctenopharyngodon idella)via the Nrf2 signaling pathway after Aeromonas hydrophila infection

Background Mannan oligosaccharides(MOS)are recommended as aquaculture additives owing to their excellent antioxidant properties.In the present study,we examined the effects of dietary MOS on the head kidney and spleen of grass carp(Ctenopharyngodon idella)with Aeromonas hydrophila infection.Methods A total of 540 grass carp were used for the study.They were administered six gradient dosages of the MOS diet(0,200,400,600,800,and 1,000 mg/kg)for 60 d.Subsequently,we performed a 14-day Aeromonas hydrophila challenge experiment.The antioxidant capacity of the head kidney and spleen were examined using spectrophotometry,DNA fragmentation,qRT-PCR,and Western blotting.Results After infection with Aeromonas hydrophila,400-600 mg/kg MOS supplementation decreased the levels of reactive oxygen species,protein carbonyl,and malonaldehyde and increased the levels of anti-superoxide anion,antihydroxyl radical,and glutathione in the head kidney and spleen of grass carp.The activities of copper-zinc superoxide dismutase,manganese superoxide dismutase,catalase,glutathione S-transferase,glutathione reductase,and glutathione peroxidase were also enhanced by supplementation with 400-600 mg/kg MOS.Furthermore,the expression of most antioxidant enzymes and their corresponding genes increased significantly with supplementation of 200-800 mg/kg MOS.mRNA and protein levels of nuclear factor erythroid 2-related factor 2 also increased following supplementation with 400-600 mg/kg MOS.In addition,supplementation with 400-600 mg/kg MOS reduced excessive apoptosis by inhibiting the death receptor pathway and mitochondrial pathway processes.Conclusions Based on the quadratic regression analysis of the above biomarkers(reactive oxygen species,malondialdehyde,and protein carbonyl)of oxidative damage in the head kidney and spleen of on-growing grass carp,the recommended MOS supplementation is 575.21,557.58,531.86,597.35,570.16,and 553.80 mg/kg,respectively.Collectively,MOS supplementation could alleviate oxidative injury in the head kidney and spleen of grass carp infected with Aeromonas hydrophila.

Enzymatically prepared alginate oligosaccharides improve broiler chicken growth performance by modulating the gut microbiota and growth hormone signals

Background Alginate oligosaccharide(AOS)holds great potential as a novel feed supplement in farm animals.However,the effects of AOS on chicken health and the underlying mechanisms are not fully understood.This study aimed to optimize the enzymatic preparation of AOS by using bacterial alginate lyases expressed in yeast,investigate the effects of the prepared AOS on the growth performance and gut health of broiler chickens,and reveal the underlying mechanisms.Results Five alginate lyases from bacteria were cloned into Pichia pastoris GS115 and the alginate lyase PDE9 was expressed at relatively high yield,activity and stability in P.pastoris.Animal trials were carried out using 3201-day-old male Arbor Acres broilers(four groups;8 replicates/group×10 chicks/replicate)receiving either a basal diet or the same diet supplemented with 100,200 and 400 mg/kg PDE9-prepared AOS for 42 d.The results showed that dietary supplementation of 200 mg/kg AOS displayed the highest activity in promoting the birds’ADG and ADFI(P<0.05).AOS ameliorated the intestinal morphology,absorption function and barrier function,as indicated by the enhanced(P<0.05)intestinal villus height,maltase activity,and the expression of PEPT,SGLT1,ZNT1,and occludin.AOS also increased serum insulin-like growth factor-1,ghrelin(P<0.05),and growth hormone(P<0.1).Moreover,the concentrations of acetate,isobutyrate,isovalerate,valerate,and total SCFAs in cecum of birds fed AOS were significantly higher than the control birds(P<0.05).Metagenomic analysis indicated that AOS modulated the chicken gut microbiota structure,function,and microbial interactions and promoted the growth of SCFAs-producing bacteria,for example,Dorea sp.002160985;SCFAs,especially acetate,were found positively correlated with the chicken growth performance and growth-related hormone signals(P<0.05).We further verified that AOS can be utilized by Dorea sp.to grow and to produce acetate in vitro.Conclusions We demonstrated that the enzymatically produced AOS effectively promoted broiler chicken growth performance by modulating the chicken gut microbiota structure and function.For the first time,we established the connections among AOS,chicken gut microbiota/SCFAs,growth hormone signals and chicken growth performance.

Multiplex genome editing targeting soybean with ultra-low anti-nutritive oligosaccharides

Soybean is the primary source of plant protein for humans.Owing to the indigestibility of the raffinose family of oligosaccharides(RFO),raffinose and stachyose are considered anti-nutritive factors in soybean seeds.Low-RFO soybean cultivars are generated by mutagenesis of RFO biosynthesis genes,but the carbohydrate profiles invite further modification to lower RFOs.This study employed a pooled multiplex genome editing approach to target four seed-specifically expressed genes mediating RFO biosynthesis,encoding three raffinose synthases(RS2,RS3,and RS4)and one stachyose synthase.In T1progeny,rs2/rs3 and rs4/sts homozygous double mutants and a rs2/rs3/rs4/sts quadruple mutant(rfo-4m)were characterized.The rs2/rs3 mutant showed reduced raffinose and stachyose contents,but the rs4/sts mutant showed only reduced stachyose in seeds.The RFO contents in the rfo-4m mutant were almost eliminated.Metabolomic analysis showed that the mutation of four RFO biosynthesis genes led to a shift of metabolic profile in the seeds,including the accumulation of several oligosaccharides-related metabolites.These mutants could contribute to precision breeding of soybean cultivars for soy food production.

Therapeutic potential and mechanism of functional oligosaccharides in inflammatory bowel disease: a review

Inflammatory bowel disease(IBD)is characterized by recurrent attacks and long courses,and the number of patients has expanded rapidly year by year.Additionally,current conventional strategies exist serious adverse effects.In this case,it is an urgent issue to find out an effective and safe treatment.Functional oligosaccharides possess safe and excellent physiological activities,and have attracted enormous attention due to their great therapeutic potential for IBD.This review emphasizes the attenuating effects of distinct functional oligosaccharides on IBD and their structure,and summarizes the main mechanisms from the aspects of regulating intestinal fl ora structure,repairing intestinal barrier,modulating immune function and mediating related signaling pathways in order to reveal the relationship between functional oligosaccharides,immune regulation,intestinal epithelial cells,gut fl ora and IBD treatment.Oligosaccharides possess excellent protective effects on IBD,and can be considered as safe and functional ingredients in the health food and pharmaceutical industry.

Combinatorial Enzyme Approach to Convert Wheat Insoluble Arabinoxylan to Bioactive Oligosaccharides

Combinatorial enzyme technology was applied for the conversion of wheat insoluble arabinoxylan to oligosaccharide structural variants. The digestive products were fractionated by Bio-Gel P4 column and screened for bioactivity. One fraction pool was observed to exhibit antimicrobial property resulting in the suppression of cell growth of the test organism ATCC 8739 E. coli. It has a MIC value of 1.5% (w/v, 35°C, 20 hr) and could be useful as a new source of prebiotics or preservatives. The present results further confirm the science and useful application of combinatorial enzyme approach.

Effects of in ovo delivered xylo-and mannan-oligosaccharides on growth performance,intestinal immunity,cecal short-chain fatty acids,and cecal microbiota of broilers

Background:This study investigated a novel in ovo feeding strategy to determine the prebiotic effects of xylo-and mannan-oligosaccharides(XOS and MOS)differing in the degree of polymerization.A total of 192 fertilized eggs were divided into 6 treatment groups:i)normal saline control(NSC),ii)xylotriose(XOS3),iii)xylotetraose(XOS4),iv)mannotriose(MOS3),v)mannotetraose(MOS4),and vi)no injection control(NIC),each containing 4 replicate trays with 8 eggs per replicate.On d 17 of incubation,3 mg of oligosaccharides(except for controls)dissolved in 0.5 mL of 0.85%normal saline were injected into the amnion of Cobb 500 broilers eggs.After hatch,the chicks were raised for 28 d under standard husbandry practices and were fed a commercial broilers diet ad libitum,and samples were collected periodically.Results:The hatchability,growth performance,and relative weights of breast,drumstick,liver,and proventriculus were not different among the treatments(P>0.05).The XOS3 injection increased the total short-chain fatty acid production at d 28 compared with both control groups(P<0.05).The villus height to crypt depth ratio was significantly higher in the XOS4 group than both controls on the hatch day(P<0.01)but were not different among any treatments on d 7 and 28(P>0.05).On the hatch day,the expression level of the CD3 gene(a T cell marker)was increased by XOS3,while the IL-10 gene(a marker of anti-inflammatory cytokine)was reduced by MOS4(P<0.05)compared with both controls.Compared with both controls,XOS3 exhibited a trend of reduction for IL-10(P=0.074).No cytokines or lymphocyte markers were affected by the treatments on d 7(P>0.05),except XOS4 increased IL-4 compared with NSC(P<0.05).The broilers in the MOS4 group had higher operational taxonomic units(OTUs)and had more differentially abundant taxa,including order Lactobacillales and family Leuconostocaceae(P<0.05)than both controls on d 28.The predictive functional profiling indicated that the linoleic acid metabolism pathway was enriched in the cecal microbiota of the XOS3 group compared with both controls(P<0.05).Conclusion:The effects of these XOS and MOS on ileal mucosa and immunity are transient,but the effects on fermentation and cecal microbiota are prolonged,and further research is warranted to determine their use as a gut health promoter in poultry.

Alginate oligosaccharide-induced intestinal morphology, barrier function and epithelium apoptosis modifications have beneficial effects on the growth performance of weaned pigs

Background: Alginate oligosaccharide(AOS), produced from alginate by alginate lyase-mediated depolymerisation, is a potential substitute for antibiotics and possesses growth-enhancing effects. Nevertheless, the mechanisms by which AOS regulates porcine growth remain to be elucidated. Therefore, we investigated the AOS-mediated changes in the growth performance of weaned pigs by determining the intestinal morphology, barrier function,as well as epithelium apoptosis.Methods: Twenty-four weaned pigs were distributed into two groups(n = 12) and received either a basal diet(control group) or the same diet supplemented with 100 mg/kg AOS. On d 15, D-xylose(0.1 g/kg body weight)was orally administrated to eight randomly selected pigs per treatment, and their serum and intestinal mucosa samples were collected 1 h later.Results: Our results showed that inclusion of AOS in the diet for 2 wk increased(P < 0.05) the average daily body weight gain in weaned pigs. Notably, AOS supplementation ameliorated the intestinal morphology and barrier function, as suggested by the enhanced(P < 0.05) intestinal villus height, secretory immunoglobulin A content and goblet cell counts. Compared to the control group, AOS ingestion both decreased(P < 0.05) the total apoptotic percentage and increased(P < 0.05) the proportion of S phase in the intestinal epithelial cells. Furthermore, AOS not only up-regulated(P < 0.05) the B-cell lymphoma-2(BCL2) transcriptional level but also down-regulated(P < 0.05) the B-cell lymphoma-2-associated X protein(BAX), cysteinyl aspartate-specific proteinase-3(caspase-3) and caspase-9 transcriptional levels in the small intestine.Conclusions: In summary, this study provides evidence that supplemental AOS beneficially affects the growth performance of weaned pigs, which may result from the improved intestinal morphology and barrier function,as well as the inhibited enterocyte death, through reducing apoptosis via mitochondria-dependent apoptosis.

Alginate oligosaccharides preparation, biological activities and their application in livestock and poultry

Alginate oligosaccharides(AOS), belonging to the class of functional marine oligosaccharides, are low-molecular polymers linked by β-1,4-mannuronic acid(M) and α-1,4-guluronic acid(G), which could be classically obtained by enzymatic hydrolysis of alginate. With low viscosity and good water solubility, as well as anti-oxidant, immune regulation, anti-bacterial and antiinflammatory activities, AOS have been widely used in medical science and functional food, green agriculture and other fields. As new bio-feed additives, AOS have broad potential applications in animal husbandry. In this review, the sources of alginate, chemical structure and preparation methods of AOS, and their biological activities and application in livestock and poultry are summarized. We expect this review could contribute to lay a foundation of application and further research for AOS in livestock and poultry.

Alginate oligosaccharide improves resistance to postharvest decay and quality in kiwifruit(Actinidia deliciosa cv. Bruno)

Kiwifruit is an extremely perishable fruit;postharvest disease and senescence during storage can reduce the fruit quality,resulting in economic loss.Considerable research effort has focused on identifying safe and cost-effective ways to preserve fresh kiwifruit.To this end,the present study investigated the effects of alginate oligosaccharide(AOS)soaking treatment on postharvest quality and disease in the‘Bruno’variety of kiwifruit.The involved physiological mechanisms were further explored.The results showed that AOS did not inhibit the growth of Botrytis cinerea in vitro,the causal agent of gray mold in kiwifruit,but reduced the incidence of gray mold and diameter of lesions of kiwifruit during storage.Kiwifruit treated with 50 mg·L-1 AOS showed a higher degree of firmness and lower soluble solid content than control fruit treated with distilled water.Moreover,AOS treatment inhibited the activity of polygalacturonase and pectinesterase,while enhancing the activity of polyphenoloxidase,l-phenylalanine ammonia lyase andβ-1,3-glucanase related to pathogen defense,and also improved total antioxidant capacity determined by the DPPH,FRAP,and ABTS methods in kiwifruit.These results indicate that 50 mg·L-1 AOS can confer disease resistance in kiwifruit during storage.

Laminarin and Laminarin Oligosaccharides Originating from Brown Algae:Preparation,Biological Activities,and Potential Applications

Brown algae is one of the three major types of marine algae and includes approximately 2000 species.It is widely dis-tributed in various seas around the world.Brown algae contain a plethora of active substances,such as polysaccharides,polyphe-nols,omega-3 fatty acids,and carotenoids.Laminarin,a type of storage carbohydrate found abundantly in brown algae,is mainly formed by glucose monomers linked byβ-1,3-glucosidic bonds and partialβ-1,6-glucosidic bonds.Laminarin and laminarin oligo-saccharides,which contain 2-10 saccharide units,have extensive biological activities,such as antitumor,antioxidant,anti-inflam-matory,and prebiotic properties.Moreover,both laminarin and laminarin oligosaccharides can be considered as ideal substrates for bioethanol production because they are composed of abundant glucose residues.Therefore,brown algae-derived laminarin and lami-narin oligosaccharides have various potential applications in the food,medicine,cosmetics,and bioenergy fields.This paper reviews the preparation methods of laminarin and laminarin oligosaccharides,as well as their biological activities and potential applications.

Polyguluronate Sulfate and Its Oligosaccharides but not Heparin Promotes FGF19/FGFR1c Signaling

Fibroblast growth factor 19(FGF19) functions as a hormone by affecting glucose metabolism. FGF19 improves glucose tolerance when overexpressed in mice with impaired glucose tolerance or diabetes. A functional cellular FGF19 receptor consists of FGF receptor(FGFR) and glycosaminoglycan complexed with either α Klotho or β Klotho. Interestingly, in mice with diet-induced diabetes, a single injection of FGF1 is enough to restore blood sugar levels to a healthy range. FGF1 binds heparin with high affinity whereas FGF19 does not, indicating that polysaccharides other than heparin might enhance FGF19/FGFR signaling. Using a FGFs/FGFR1 c signaling-dependent Ba F3 cell proliferation assay, we discovered that polyguluronate sulfate(PGS) and its oligosaccharides, PGS12 and PGS25, but not polyguluronate(PG), a natural marine polysaccharide, enhanced FGF19/FGFR1 c signaling better than that of heparin based on ~3H-thymidine incorporation. Interestingly, PGS6, PGS8, PGS10, PGS12, PGS25, and PGS, but not PG, had comparable FGF1/FGFR1 c signal-stimulating activity compared to that of heparin. These results indicated that PGS and its oligosaccharides were excellent FGF1/FGFR1 c and FGF19/FGFR1 c signaling enhancers at cellular level. Since the inexpensive PGS and PGS oligosaccharides can be absorbed through oral route, these seaweed-derived compounds merit further investigation as novel agents for the treatment of type 2 diabetes through enhancing FGF1/FGFR1 c and FGF19/FGFR1 c signaling in future.

Human Milk Oligosaccharides Enhance Innate Immunity to Respiratory Syncytial Virus and Influenza <i>in Vitro</i>

Human milk oligosaccharides (HMO) contribute to innate immunity by enhancing growth of beneficial bacteria, epithelial cell maturation and mucosal barrier integrity. They have immunomodulatory effects and can block pathogen binding to host cell surface glycans or receptors. We investigated the effects of 2’-fucosyllactose (2’FL), 6’-sialyllactose (6’SL), 3’-sialyllactose (3’SL) and lacto-N-neoTetraose (LNnT) on human respiratory epithelial cell lines or peripheral blood mononuclear cells (PBMCs) following respiratory viral infectionin vitro. Expression of cytokines and viral load were monitored in infected cells. These biomarkers of innate immunity were selected since viral load and cytokine levels (IP-10, MIP-1α, IL-6, IL-8, TNF-α) have been correlated with disease severity in respiratory syncytial virus (RSV) and influenza (IAV) virus infectionin vivo. 2’FL significantly decreased RSV viral load and cytokines associated with disease severity (IL-6, IL-8, MIP-1α) and inflammation (TNF-α, MCP-1) in airway epithelial cells. LNnT and 6’SL significantly decreased IAV viral load in airway epithelial cells. 6’SL dose-dependently down-regulated IP-10 and TNF-α in RSV infected PBMCs. HMO at or below levels found in breast milk enhance innate immunity to respiratory viruses in vitro and may interact directly with cells to modulate biomarkers of innate immunity.

Individual and Combined Effects of Nucleotides and Human Milk Oligosaccharides on Proliferation, Apoptosis and Necrosis in a Human Fetal Intestinal Cell Line

Nucleotides (NT) and human milk oligosaccharides (HMO) individually affect epithelial cell growth, but their combined effects had not been studied. Herein, the impact of NT and HMO on cell proliferation, apoptosis, necrosis and cell cycle in the fetal epithelial cell line (FHs-74 Int) was determined. Cells were incubated with media containing 2.5% FBS and no epidermal growth factor (Control);fucosyllactose (FL) mix [85% 2’FL/15% 3’FL], sialyllactose (SL) mix [40% 6’SL/10% 3’SL/50% sialic acid (SA)] or LNnT at 125, 250, 500 or 1000 μg/mL with and without 250 μg/mL NT (43% CMP, 18.5% UMP, 16.4% AMP, and 22.0% GMP) for 24 or 72 h. NT alone significantly increased proliferation, but did not affect cell cycle or apoptosis/necrosis. All HMO treatments at 1000 μg/mL significantly decreased proliferation and some were also inhibitory at 250 or 500 μg/mL. When NT and HMO were simultaneously added, NT ameliorated the anti-proliferative effect of HMO. FL significantly increased cells in S phase and SL and LNnT treatments significantly increased cells in G2/M and S phases, which concomitantly decreased cells in G0/G1. HMO with NT significantly decreased the percent of cells in the G2/M phase compared to HMO alone. Higher HMO doses significantly increased the percentage of apoptotic and necrotic cells compared to control. In conclusion, HMO reduced cell proliferation and this effect is partially ameliorated by NT. It appears that HMO initially induced apoptosis/necrosis, which was later evidenced by G2/M cell cycle arrest and decreased proliferation.

Sucrose-free hawthorn leathers formulated with fructooligosaccharides and xylooligosaccharides ameliorate high-fat diet induced inflammation, glucose and lipid metabolism in liver of mice

High sucrose content in traditional hawthorn leathers limits the potential consumption, particularly for elders and diabetics. In this study, sucrose-free hawthorn leathers were formulated with 75% fructooligosaccharides(FOS) and 25% xylooligosaccharides(XOS)(HLF75), which exhibited comparable morphology and sensory properties to the traditional ones. Then, the anti-obesity activity of HLF75 was investigated using high-fat diet(HFD) fed C57BL/6J mice. Comparing with traditional hawthorn leathers, HLF75 supplementation in HFD significantly decreased the levels of blood glucose and serum lipid. The histomorphologies of liver and subcutaneous fat tissues were ameliorated by HLF75, as well as the down-regulated m RNA expression levels of IL-1β, Nos2 and Cox-2 in the liver. M oreover, the protein levels of M y D88 and NF-κB in the liver were suppressed by HLF75 treatment with decreased F4/80-positive macrophage number. Ho wever, the expression levels of PI3K, phosphorylated-AKT(Thr308), and phosphorylated-m TOR(Ser2448) proteins related to glucose metabolism were increased in the liver. Moreover, fat synthesis-related gene expression in HLF75-fed mice was suppressed while expressions of lipolysis genes were improved. Thus, HLF75 supplementation alleviated HFD-induced obesity through the alleviation of inflammation and restoration of the disturbed glucose and lipid metabolism. Functional oligosaccharides could be effective sucrose substitutes in hawthorn leathers and enable their potential utilization as functional foods.

Combinatorial Enzyme Approach to Produce Oligosaccharides of Diverse Structures for Functional Screen

Combinatorial chemistry has been a focus of research activity in modern drug discovery and biotechnology. It is a concept by which a vast library of molecular diversity is synthesized and screened for target properties. This report is to illustrate the application of enzyme technology using the concept of combinatorial chemistry as a novel approach for the bioconversion of plant fibers. Citrus pectin was subjected to combinatorial enzyme digestion to create libraries of pectic oligosaccharides with diverse structural variants. Repeated cycles of fractionation and screening resulted in the isolation and identification of an active oligoGalA species with antimicrobial activity.

Comparative Analysis of Lactulose and Fructooligosaccharide on Growth Kinetics, Fermentation, and Antioxidant Activity of Common Probiotics

Prebiotics are non-digestible oligosaccharides that selectively stimulate the growth of beneficial bacteria in the human gut. Fructooligosaccharide (FOS) is a common prebiotic found in food products and infant formula. Lactulose is primarily used as a pharmaceutical ingredient but also shows potential prebiotic activities. Our objectives were to determine and compare the effects of FOS and lactulose on: 1) growth kinetics of common probiotics in aerobic condition;2) pH and titratable acidity after fermentation;and 3) antioxidant capacity of the probiotics. Ten probiotic and two non-probiotic strains, representing genera Lactobacillus, Bifidobacterium, Bacillus, and Escherichia were assembled. Media used for prebiotics experiment were modified to contain 2% FOS or lactulose as the sole or main carbohydrate source. All experiments were done in triplicate. In aerobic condition, most strains cultured with FOS or lactulose did not grow optimally compared to dextrose (a non-prebiotic), while all four Bifidobacterium spp. showed little growth regardless of the carbohydrate source. In anaerobic condition, lactulose and FOS fermentation of Bifidobacterium spp. yielded similar pH (p = 0.2723), but percent lactic acid, as determined by titratable acidity, was higher after lactulose fermentation (p = 0.0004). The non-probiotic strains were able to utilize both FOS and lactulose, but displayed weaker acid production and higher pH (p Bifidobacterium spp. (p = 0.0002) and Lactobacillus spp. (p = 0.0447), but not probiotic E. coli and Bacillus spp. (p = 0.2599) or non-probiotics (p = 0.8816). In conclusion, lactulose supported growth activities of probiotics to a similar extent as FOS. Lactulose also stimulated higher acid production for Bifidobacterium spp. than FOS in anaerobic condition, thus it might be considered for incorporation into functional food products containing bifidobacteria.

Antimicrobial Activity of Eucosterol Oligosaccharides Isolated from Bulb of Squill (<i>Scilla scilloides</i>)

Antimicrobial activity of methanol extract from bulb of Scilla scilloides as well as the purified eucosterol oligosaccharides (EOs) against bacteria, fungi and alga was evaluated in vitro using paper disc bioassay. EOs including scillascilloside E-1, E-2, E-3 and G-1 were isolated from the bulbs by methanol extraction, gel filtration on Sephadex LH-20 and preparative HPLC. The metabolites were identified by MS (HR-FAB-MS) and 1H- and 13C-NMR spectral data analyses. Methanol (MeOH) extract and purified compounds, EOs showed a selective inhibitory activity against eukaryotic cells including fungal species such as Aspergillus flavus, Candida albicans, Pyricularia oryzae and an alga such as Chlorella regulsris at the concentration of 200 μg/paper disc, but little active against bacteria. Out of four EO compounds, Scilla-scilloside E-3 revealed the highest activity. These results show that the MeOH extract and EOs from the medicinal plant, S. scilloides, may be applied as a natural fungicide or a food preservative for control of molds.