Chemical characterization of the oligosaccharides in Bactrian camel (Camelus bactrianus) milk and colostrum

Bactrian camel milk and colostrum are commonly used as foods in Mongolia, whose people believe that these products promote human health. It has been hypothesized that milk oligosaccharides are biologically significant components of human milk, acting as receptor analogs that inhibit the attachment of pathogenic microorganisms to the colonic mucosa, and as prebiotics, which stimulate the growth of bifidobacteria within the infant colon. To evaluate their biological significance, we studied the oligosaccharides present in samples of Bactrian camel milk and colostrum. Using ¹H-nuclear magnetic resonance spectroscopy, we identified and characterized the following oligosaccharides of camel colostrum: Gal(β1–4)[Fuc(α1–3)]Glc (3-fucosyllactose), Gal(β1–3)Gal(β1–4)Glc (3′-galactosyllactose), Gal(β1–6)Gal(β1–4)Glc (6′-galactosyllactose), Neu5Ac(α2–3)Gal(β1–4)Glc (3′-sialyllactose), Neu5Ac(α2–6)Gal(β1–4)Glc (6′-sialyllactose), Neu5Ac(α2–3)Gal(β1–3)Gal(β1–4)Glc (sialyl-3′-galactosyllactose), Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc (sialyllacto-N-tetraose c), Neu5Ac(α2–3)Gal(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (sialyllacto-N-novopentaose a), Gal(β1–3)[Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (sialyllacto-N-novopentaose b); and Neu5Ac(α2–6)Gal(β1–4)GlcNAc(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (monosialyllacto-N-neohexaose). The oligosaccharides in the mature camel milk were characterized as 3′-galactosyllactose, Gal(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (lacto-N-novopentaose I), and 3′-sialyllactose.     

Chemical characterisation of saccharides in the milk of a reindeer (Rangifer tarandus tarandus)

Although reindeer milk is utilised as a food in some areas in the world, including Mongolia, the saccharides in this milk have thus far not been explored. In this study, the structures of the separated saccharides were characterised by ¹H-nuclear magnetic resonance spectroscopy to be as follows: Gal(β1-3)Gal(β1-4)Glc (3′-galactosyllactose, 3′-GL), Gal(β1-6)Gal(β1-4)Glc (6′-galactosyllactose, 6′-GL), Gal(β1-4)Glc (lactose), Neu5Ac(α2-6)Gal(β1-4)Glc (6′-N-acetylneuraminyllactose, 6′-SL), Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-3)Gal(β1-4)Glc (sialyllacto-N-tetraose c, LST c) and uridine 5′-diphospho-α-N-acetyl-D-glucosamine (UDP-GlcNAc). In addition, monosaccharide-α-1-phosphate and some other UDP-mono or oligosaccharides were found, but Neu5Ac(α2-3)Gal(β1-4)Glc (3′-N-acetylneuraminyllactose, 3′-SL) was not detected. Lactose was the predominant saccharide. It may be concluded that, in this milk, the concentration of phosphorylated or UDP-saccharides was higher than that of Neu5Ac containing oligosaccharides; this should be a significant feature of this species milk.     

Concentrations of sialyloligosaccharides in bovine colostrum and milk during the prepartum and early lactation

Sialyloligosaccharides and sialylglycoconjugates in colostrum and milk are regarded to be important biological components with respect to be source of brain gangliosides in infant and to be antiinfectional components for the attack by the pathogenic bacteria and virus. Several acidic oligosaccharides have been characterised in both bovine and human milk or colostrum. The sialyloligosaccharide content of human colostrum and milk has been extensively studied, whereas that of cows milk and colostrum has received less attention. In this study, the concentrations of three sialyloligosaccharides of bovine colostrum and milk were determined at various stages during the prepartum and the first 7 d postpartum. The concentration of 3'SL (Neu5Ac(alpha2-3)Gal(beta1-4)Glc) reached a maximum value of 0.85 mg/ml immediately following parturition while the concentrations of 6'SL (Neu5Ac(alpha2-6)Gal(beta1-4)Glc) and 6'SLN (Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc) of 0.14 and 0.12 mg/ml, respectively, were much lower at this initial stage, although these concentration were maximum immediately following parturition. Bovine colostrum, especially that collected immediately after parturition, may be suitable as a source of 3'SL and other sialyloligosaccharides for use as additives by the food or pharmaceutical industries.     

Qualitative and quantitative mass spectrometry comparison of characteristic galactosyl lactose isomers from goat milk at different lactation stages

Galactooligosaccharides are composed mainly of galactosyl lactose, which is important for infant growth and as a functional food additive. Although galactosyl lactose is abundant in goat milk, its complex structure has hindered the separation and analysis of its isomers. In this study, 5 isomers of goat milk galactosyl lactose were separated by HPLC: β6′-galactosyl lactose (β6′-GL), α6′-galactosyl lactose (α6′-GL), β4′-galactosyl lactose (β4′-GL), α3′-galactosyl lactose (α3′-GL), and β3′-galactosyl lactose (β3′-GL). This composition differs from that of commercial galactooligosaccharide products, which comprise mainly β-configuration oligosaccharides. The isomers were then qualitatively and quantitatively compared at different lactation stages using online HPLC-mass spectrometry. Relative quantitative analysis showed that the total content of the 5 galactosyl lactose isomers was highest in transitional goat milk. Specifically, β3′-GL was the main isomer in colostrum and α3′-GL was the main isomer in transitional and mature milk. β6′-Galactosyl lactose and β4′-GL tended to increase and then decrease during lactation. Moreover, α3′-GL content was 2 times higher than in colostrum and 10 times higher in transitional milk than in mature milk; in contrast, for β3′-GL, the values were 5 and 2 times higher, respectively. Absolute quantitative analysis revealed that β3′-GL was the most abundant isomers in colostrum (32.3 mg/L), and α3′-GL was the most abundant in transitional milk (88.1 mg/L) and mature milk (36.3 mg/L). These findings provide an important quantitative basis for understanding the relationship between structure and function of galactosyl lactose in goat milk, as well as its exploitation as a functional food.     

Identification of Sialylated Oligosaccharides Derived from Ovine and Caprine Caseinomacropeptide by Graphitized Carbon Liquid Chromatography–Electrospray Ionization Ion Trap Tandem Mass Spectrometry

Isolation and characterization of oligosaccharides from caseinomacropeptide (CMP) are important in understanding the biological and functional properties of CMP. However, it is difficult to achieve this goal, due to the high degree of isomerism present in these types of compounds. In this study, the sialylated oligosaccharides derived from ovine and caprine CMP were released as oligosaccharide alditols by reductive β-elimination and subsequently separated and characterized using graphite carbon column liquid chromatography–negative electrospray ionization ion trap tandem mass spectrometry (LC/ESI(?)-MSⁿ). Although, the chromatographic resolution of isomeric oligosaccharides was not achieved perfectly, the characteristic tandem mass spectra of these compounds allowed differentiating and confirming unequivocally the structure of each one of the oligosaccharides. In CMP of both species, four trisaccharides and four tetrasaccharides were identified as O-glycans. Their chemical structures were identified to be Galβ1-3(NeuAcα2-6)GalNAcol, NeuAcα2-3Galβ1-3GalNAcol, Galβ1-3(NeuGcα2-6)GalNAcol, NeuGcα2-3Galβ1-3GalNAcol, NeuAcα2-3Galβ1-3(NeuGcα2-6)GalNAcol, NeuGcα2-3Galβ1-3(NeuAcα2-6)GalNAcol, NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAcol, and NeuGcα2-3Galβ1-3(NeuGcα2-6)GalNAcol. The LC/MSⁿ methodology using an ion trap-type mass analyzer shown in this study is of general applicability for determination of short O-glycan oligosaccharides.     

Rapid,quantitative analysis of 3′- and 6′-sialyllactose in milk by flow-injection analysis–mass spectrometry: Screening of milks for naturally elevated sialyllactose concentration

Non-protein-bound oligosaccharides are important bioactive components of cow milk, with potential human-health benefits such as stimulation of the growth of beneficial gut bacteria and defense against pathogens. In bovine milk, the majority of oligosaccharides are sialylated; 3′-sialyllactose (3′-N-acetylneuraminyl-d-lactose; 3′-SL) is the predominant sialylated oligosaccharide, followed by 6′-sialyllactose (6′-N-acetylneuraminyl-d-lactose; 6′-SL). Both 3′-SL and 6′-SL have antimicrobial activity. As bovine milk products such as infant formula can be an important component of the human diet, and the concentrations of 3′-SL and 6′-SL are lower in bovine milk compared with human milk, we aimed to identify cows that naturally produce higher concentrations of sialyllactose in their milk. Milk from such cows could be used to produce foods with an increased sialyllactose content, potentially providing increased health benefits. We speculated that cows overexpressing 3′-SL and 6′-SL would exist at low frequency in the population and, to allow their efficient identification, we developed a novel assay for 3′-SL and 6′-SL utilizing flow-injection analysis-mass spectrometry, which could be used for high-throughput analysis of milk samples. We then determined 3′-SL and 6′-SL concentrations in milk samples from 15,507 cows from Friesian, Jersey, and Friesian-Jersey crossbred animals. We found 329 cows with concentrations of 3′-SL or 6′-SL >2-fold higher than the mean, 26 cows with concentrations of 3′-SL or 6′-SL >3-fold higher than the mean, and 1 cow with concentrations of 3′-SL >4-fold higher than the mean. Although these outliers were observed across the 3 groups of cows, breed had a strong effect on mean 3′-SL and 6′-SL concentrations.     

鹿蹄橐吾多糖LW21的分离纯化及其结构分析

目的:确定鹿蹄橐吾多糖LW21的结构,为探讨鹿蹄橐吾多糖的药理活性,合理利用鹿蹄橐吾这种植物资源提供依据。方法:水提取醇沉获得鹿蹄橐吾水溶性粗多糖LW,经酸性乙醇分级和DEAE-SephdexA-25纯化得多糖LW21。纸层析、醋酸纤维薄膜电泳和Sepharose CL-4B柱层析进行纯度鉴定,LW21结构分析采用高碘酸氧化、Smith降解、甲基化分析及IR、NMR、GC和GC-MS等方法。结果表明:LW21为均一多糖,相对分子质量约为1.1×106。其单糖组成为鼠李糖(Rha)、阿拉伯糖(Ara)、甘露糖(Man)、葡萄糖(Glc)、半乳糖(Gal),物质的量比为7.4:11.9:25.7:40.0:14.9。多糖LW21为有分枝结构,主链由Glc和Man构成,其中其中Man主要以β(1→2)和β(1→6)糖苷键连接,β(1→2)糖苷键在3-O处和6-O处有分枝,β(1→6)糖苷键在2-O和4-O处有分枝,Glc也主要以β(1→2)及β(1→6)糖苷键连接,β(1→2)糖苷键在6-O处有分枝,β(1→6)糖苷键连接,在2-O处有分枝。分子支链由Ara、Rha、Gal构成。末端残基为Gal、Ara、Rha、Glc。结论:鹿蹄橐吾多糖LW2 1是一新结构多糖,为首次从鹿蹄橐吾中分离得到。     

气相色谱法同时测定多糖中的中性糖、糖醛酸、氨基糖和唾液酸

采用甲醇解和硅烷衍生化,用气相色谱法同时测定样品中的中性糖、糖醛酸、N-乙酰氨基糖和唾液酸,取得了很好的效果。3种标准单糖混合物(包含阿拉伯糖、鼠李糖、岩藻糖、木糖、甘露糖、半乳糖、葡萄糖、葡萄糖醛酸、半乳糖醛酸、N-乙酰半乳糖、N-乙酰葡萄糖、5-乙酰唾液酸)和2种多糖样品(豆腐渣多糖DFP、鸡腿菇多糖F32)用1 mol/L盐酸甲醇在85℃反应18～24 h,碳酸银中和,加入乙酸酐室温暗处反应24 h,使氨基糖重新引入N-乙酰基,除银盐,干燥,加入硅烷化试剂[V(吡啶)∶V(六甲基二硅氨烷)∶V(三甲基氯硅烷)=5∶1∶1),室温放置30 min,最后用气相色谱进行分析。     

N-丙酰-唾液酸衍生物的化学酶法合成及在测定禽蛋中唾液酸含量的应用

为实现禽类蛋黄和蛋清中N-乙酰神经氨酸(N-acetylneuraminic acid,Neu5Ac)的准确定量,消除禽蛋样品中分析物以外的物质产生的基质效应对分析结果造成的影响,本研究以甘露糖胺为底物采用化学酶法合成了非天然唾液酸衍生物5-溴-4-氯-3-吲哚-β-D-半乳糖苷-N-丙酰-唾液酸(5-Bromo-4-...     

Changes in chemical composition of Alxa bactrian camel milk during lactation

Changes in chemical composition of Alxa bactrian camels reared in Inner Mongolia (China) during lactation were investigated. Colostrum and milk samples from 10 nomadic female camels in their first season of lactation were collected periodically from parturition until 90 d postpartum (PP). The average contents of gross composition were 14.23% protein, 4.44% lactose, 0.27% fat, 0.77% ash, and 20.16% total solids in colostrum at 2 h PP, and the respective mean values were 3.55, 4.24, 5.65, 0.87, and 14.31% for regular milk on d 90. A 15-fold increase was shown in fat content during the first 24 h, whereas a sharp decrease was shown during the first 12 h of lactation in protein, ash, and total solids contents. Variation in lactose content was small (4.24 to 4.71%) throughout the study period. Total N, nonprotein N, casein N, and whey protein N were found to be 2.23, 0.06, 0.86, and 1.31 g/100 mL for the colostrum at 2 h PP; and 0.56, 0.04, 0.45, and 0.07 g/100 mL for the milk at 90 d PP. Percentages of caseins increased steadily, whereas whey proteins declined gradually until 3 mo of lactation. Gas liquid chromatography analysis of milk fat showed that the content of even-numbered saturated fatty acids (C_12:0-C_18:0) in camel colostrum (2 h to 7 d PP) was lower than that of regular milk (15 to 90 d PP). The predominant saturated fatty acids were C_14:0, C_16:0, and C_18:0, regardless of the stage of lactation. There was a considerable level of polyunsaturated fatty acids (mainly C_18:1) in Alxa camel's milk fat. The levels of Ca, P, Na, K, and Cl were 222.58, 153.74, 65.0, 136.5, and 141.1 mg/100 g, respectively, at 2 h PP; the values of the minerals were 154.57, 116.82, 72.0, 191.0, and 152.0 mg/100 g, respectively, for the regular milk on d 90. The levels of vitamins A, C, E, B₁, B₂, B₆, and D were 0.97, 29.60, 1.45, 0.12, 1.24, 0.54 mg/L, and 640 IU/L, respectively, in Alxa camel milk at 90 d PP. Vitamin A and C contents were higher and vitamins E and B₁ were lower than those in colostrum. Sodium dodecyl sulfate-PAGE and densitometry results demonstrated that Alxa camel colostrum is rich in immunoglobulins, serum albumin, and 2 unknown fractions, which are reduced in amount (%) within 2 d of lactation. It seems that there is lack of β-lactoglobulin in Alxa camel milk, whereas casein and α-lactalbumin start at a low level and increase gradually until they reach their regular levels in the milk.     

Effects of cooling and freezing storage on the stability of bioactive factors in human colostrum

Breast milk constitutes the best form of newborn alimentation because of its nutritional and immunological properties. Banked human milk is stored at low temperature, which may produce losses of some bioactive milk components. During lactation, colostrum provides the requirements of the newborn during the first days of life. The aim of this study was to evaluate the effect of cooling storage at 4°C and freezing storage at -20°C and -80°C on bioactive factors in human colostrum. For this purpose, the content of IgA, growth factors such as epidermal growth factor, transforming growth factor (TGF)-β1 and TGF-β2, and some cytokines such as IL-6, IL-8, IL-10, and tumor necrosis factor (TNF)-α, and its type I receptor TNF-RI, were quantified. Some colostrum samples were stored for 6, 12, 24, and 48 h at 4°C and others were frozen at -20°C or -80°C for 6 and 12 mo. We quantified IgA, epidermal growth factor, TGF-β1, and TGF-β2 by indirect ELISA. Concentrations of IL-6, IL-10, and TNF-α cytokines, IL-8 chemokine, and TNF-RI were measured using the BD Cytometric Bead Array (BD Biosciences, Erembodegem, Belgium). Bioactive immunological factors measured in this study were retained in colostrum after cooling storage at 4°C for at least 48h, with the exception of IL-10. None of the initial bioactive factor concentrations was modified after 6 mo of freezing storage at either -20°C or -80°C. However, freezing storage of colostrum at -20°C and -80°C for 12 mo produced a decrease in the concentrations of IgA, IL-8, and TGF-β1. In summary, colostrum can be stored at 4°C for up to 48 h or at -20°C or -80°C for at least 6 mo without losing its immunological properties. Future studies are necessary to develop quality assurance guidelines for the storage of colostrum in human milk banks, and to focus not only on the microbiological safety but also on the maintenance of the immunological properties of colostrum.     

牛免疫球蛋白G的体外人源唾液酸化及Fc片段的制备

建立将牛免疫球蛋白G (bovine immunoglobulin G,bIgG)糖链末端N-羟乙酰神经氨酸酶切并连接人源N-乙酰神经氨酸(N-acetylneuraminic acid,Neu5Ac)的方法,在实现bIgG转化为人源IgG (human IgG,hIgG)的基础上,研究hIgG可结晶(Fc)片段的制备...     

Antioxidant activities of enzymatic‐hydrolysed proteins of dromedary (Camelus dromedarius) colostrum

This work investigated the antioxidant activities of dromedary colostrum proteins before and after hydrolysis by pepsin, trypsin, α‐chymotrypsin, pancreatin and papain. The enzymatic hydrolysis affected the degrees of hydrolysis, electrophoretic profiles, molecular weight distribution and hydrophobic/hydrophilic properties of the generated peptides. The antioxidant activities were evaluated using four antioxidant assays, including 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) radical‐scavenging activities, ferric reducing power and ferrous ion chelating activity. Interestingly, the antioxidant activities of dromedary colostrum proteins were enhanced after enzymatic hydrolysis. The highest antioxidant potential was obtained by pancreatic hydrolysates (P ≤ 0.05). These results suggest that dromedary colostrum protein hydrolysates are an important source of natural antioxidant peptides.     

骆驼乳对小鼠酒精性肝损伤的保护作用

目的:研究骆驼乳对Lieber-DeCarli液体饲料诱导的小鼠酒精性肝损伤的保护作用.方法:将40只雄性C57BL/6NCr小鼠随机分为对照组(Con)、模型组(Et)、骆驼乳高剂量组(EtCM_H,3 g/kg)、骆驼乳低剂量组(EtCM_L,1.5 g/kg)和阳性对照组(美他多辛,300 mg/kg).试验8周...     

Human milk oligosaccharide consumption by probiotic and human-associated bifidobacteria and lactobacilli

Human milk contains high concentrations of nondigestible complex oligosaccharides (human milk oligosaccharides; HMO) that reach the colon and are subsequently fermented by the infant gut microbiota. Using a high-throughput, low-volume growth determination, we evaluated the ability of 12 lactobacilli and 12 bifidobacteria strains, including several commercial probiotics, to ferment HMO and their constituent monomers. Of the 24 strains tested, only Bifidobacterium longum ssp. infantis ATCC 15697 and Bifidobacterium infantis M-63 were able to ferment 3′-sialyllactose, 6′-sialyllactose, 2′-fucosyllactose, and 3′-fucosyllactose. Bifidobacterium infantis M-63 degraded almost 90% of the 2′-fucosyllactose but left most of the fucose in the supernatant, as detected by HPLC. Among bifidobacteria, only the B. infantis strains and Bifidobacterium breve ATCC 15700 were able to ferment lacto-N-neotetraose (LNnT). Among lactobacilli, Lactobacillus acidophilus NCFM was found to be the most efficient at utilizing LNnT. The extracellular β-galactosidase (lacL, LBA1467) of L. acidophilus NCFM cleaves the terminal galactose of LNnT for growth, leaving lacto-N-triose II in the media as detected by HPLC. Inactivation of lacL abolishes growth of L. acidophilus NCFM on LNnT. These results contribute to our knowledge of HMO–microbe interactions and demonstrate the potential for synbiotic combinations of pre- and probiotics.     

Composition of milk from minor dairy animals and buffalo breeds: a biodiversity perspective

A comprehensive review is presented of the nutrient composition for buffalo, mare, and dromedary camel milks at the level of breed, and species‐level data for yak, mithun, musk ox, donkey, Bactrian camel, llama, alpaca, reindeer and moose milks. Average values of nutrients were calculated and compared. Interspecies values (g 100 g^?1) were 0.7–16.1 for total fat, 1.6–10.5 for protein, 2.6–6.6 for lactose, and 67.9–90.8 for water. Reindeer and moose milks had the highest fat and protein concentrations and the lowest lactose contents. Mare and donkey milks had the lowest protein and fat contents, in addition to showing the most appropriate fatty acid profile for human nutrition. Dromedary camel milk was most similar to cow milk in proximate composition. Moose milk was the richest in minerals, having values as high as 358 mg 100 g^?1 for calcium, 158 mg 100 g^?1 for sodium and 150 mg 100 g^?1 for phosphorus. Interbreed differences of 4 g 100 g^?1 were observed in total fat in buffalo, yak, mare and dromedary camel milks. Large interbreed differences were also present in the mineral contents in mare, buffalo and dromedary camel milks. By bringing together these compositional data, we hope to usefully widen the biodiversity knowledge base, which may contribute to the conservation and sustainable use of milk from underutilized dairy breeds and species, and to improved food and nutrition security, particularly in developing countries. Copyright © 2011 Society of Chemical Industry     

Short communication: The effect of heat treatment of bovine colostrum on the concentration of oligosaccharides in colostrum and in the intestine of neonatal male Holstein calves

The objective of this study was to determine the effect of the heat treatment (HT, 60°C for 60 min) on the concentration of bovine colostrum oligosaccharides (bCO) in pooled bovine colostrum and the intestine of neonatal male Holstein calves after feeding. First-milking colostrum was pooled from both primiparous and multiparous cows, and half of the pooled colostrum was heat-treated at 60°C for 60 min (HC), whereas the other half was not heat-treated and remained fresh (FC). At birth, 32 male Holstein calves were randomly assigned to 1 of 3 treatment groups: (1) control calves that did not receive colostrum for the duration of the experiment and were euthanized at 6 h (NC, n = 4) or 12 h (NC, n = 4), (2) calves fed fresh colostrum (FC) and were euthanized at 6 h (FC, n = 6) or 12 h (FC, n = 6), or (3) calves fed heat-treated colostrum (HC) and euthanized at 6 h (HC, n = 6) or 12 h (HC, n = 6). All calves were fed 2 L of colostrum within 1 h after birth. At dissection, digesta of the distal jejunum, ileum, and colon was collected and analyzed by liquid chromatography-mass spectrometry to determine the concentration of bCO within each intestinal region. The heat-treated colostrum displayed numerically higher concentrations of total bCO (3,511.6 μg/g) when compared with fresh colostrum (1,329.9 μg/g), with 3′-sialyllactose being the most abundant bCO in both fresh and HT colostrum. In contrast, calves fed HT colostrum displayed a lower amount of total bCO in the distal jejunum (221.91 ± 105.3 vs. 611.26 ± 265.1 μg/g), ileum (64.97 ± 48.39 vs. 344.04 ± 216.87 μg/g), and colon (25.60 ± 13.1 vs. 267.04 ± 125.81 μg/g) at 6 h of life when compared with calves fed fresh colostrum. No differences were observed in regard to the concentrations of total bCO in the intestine of FC and HC calves at 12 h of life. It is speculated that lower concentrations of bCO in the gastrointestinal tract of HC calves at 6 h of life could be due to the early establishment of beneficial bacteria, such as Bifidobacterium, in HC calves and their subsequent metabolism of bCO as a carbon source. These findings suggest that the heat treatment of colostrum increases the amount of free bCO, which may serve as prebiotics available to microbiota within the intestine of the neonatal calf.     

Dietary intervention with sialylated lactulose affects the immunomodulatory activities of mice

Four sialylated lactuloses [N-acetylneuraminic acid-α2,3-lactulose (Neu5Acα2,3lactulose), N-acetylneuraminic acid-α2,6-lactulose (Neu5Acα2,6lactulose), deaminoneuraminc acid-α2,3-lactulose (Kdnα2,3lactulose), and deaminoneuraminc acid-α-2,6-lactulose (Kdnα2,6lactulose)] were reported to modulate the immunity of mice. The influences of cytokine expression, cell immunity, humoral immunity, and nonspecific immunity were investigated in our study using several techniques. Analysis via ELISA showed that cytokine expression was induced by sialylated lactulose treatment consistently in the serum and spleen. Among the 4 tested sialylated lactuloses, Neu5Acα2,6lactulose performed the best, simultaneously and appropriately promoting the expression of proinflammatory and anti-inflammatory factors in the serum and spleen. Kdnα2,3lactulose showed the best antioxidant activity according to detection of the activity of superoxide dismutase, myeloperoxidase, peroxidase, and alkaline phosphatase. Flow cytometry revealed that only Kdnα2,3lactulose significantly boosted the CD3⁺ T lymphocyte ratio similarly to that of lactulose. Analysis of the hemolysin content to characterize humoral immunity revealed that Kdnα2,3lactulose notably increased hemolysin content compared with that in the control group. To evaluate the nonspecific immune effects of the 4 sialylated lactuloses, a fluorescence microsphere phagocytosis assay was used to analyze the phagocytosis of macrophages. Kdnα2,3lactulose still performed the best in enhancing the phagocytosis of macrophages, showing markedly increased phagocytic percentage and phagocytic index values compared with those in the control and lactulose groups. Comparing the differences of these 4 sialylated lactuloses in affecting immunity in mice revealed that Kdnα2,3lactulose had the best overall performance in influencing cytokine expression, cell immunity, humoral immunity, and nonspecific immunity. This study provides critical support for use of sialylated lactuloses as potential immunomodulators in foods.     

Arabidopsis β1,2-xylosyltransferase: substrate specificity and participation in the plant-specific N-glycosylation pathway

β1,2-Xylosyltransferase (XYLT) is a plant-specific glycosyltransferase that contributes to the biosynthesis of N-glycoproteins in plants. However, the specificity of XYLT for N-glycans has not yet been completely clarified. To gain insights into the function of XYLT in the plant N-glycosylation pathway, we examined the acceptor substrate specificity of recombinant Arabidopsis XYLT (AtXYLT) using 2-aminopyridine-labeled N-glycans as the substrates and confirmed the N-glycans of Arabidopsis xylt mutant. Recombinant AtXYLT expressed in insect cells required the β1,2-linked N-acetylglucosamine (GlcNAc) residue at the nonreducing terminus of the α1,3-branched mannose (Man) residue (GlcNAcβ1,2-Manα1,3-Man; GNM3B) for activity. However, AtXYLT showed decreased activity with substrates that contained α1,3-fucose at the chitobiose core-GlcNAc or a terminal GlcNAc at the α1,6-branched Man residue of GlcNAcβ1,2-Man (GlcNAcβ1,2-Manα1,6-Man; GNM3A), whose ratios were 10% and 50% of the optimal substrate, GNM3B, respectively. Moreover, AtXYLT did not show any activity in the transfer of the Xyl residue to N-glycans that contained a mammalian-type β1,4-linked galactose (Gal) residue at the nonreducing terminus of GlcNAcβ1,2-Man. These results indicate that a β1,2-linked GlcNAc residue at the nonreducing terminus of an α1,3-branched Man residue is necessary for AtXYLT activity and that mammalian-type β1,4-linked Gal residue(s) on the same branch completely inhibit(s) the activity. Furthermore, N-glycan analysis showed that approximately 30% of the N-glycans carry the Xyl residue in the wild type. These findings suggest that AtXYLT acts on protein-bound N-glycans prior to α1,3-fucosyltransferase and mannosidase II in planta.     

The Behaviour and Identification of a Range of Starch and Starch-Related Oligosaccharides Series on Gel Permeation Chromatography as a Function of Their Molecular Shapes

The elution behaviour on gel permeation chromatography of a wide range of oligosaccharides which differ in glycosidic linkage has been investigated using a high resolution column of Bio-Gel P2 (-400 mesh). The elution behaviour could be explained on size exclusion mechanisms resulting from the secondary and tertiary structures of oligosaccharides brought about by the nature of the glycosidic linkages concerned. The various series of oligosaccharides followed an order in which starch-derived materials were more included in the gel than oligosaccharides derived from other polysaccharides. The overall order was cyclomalto- < malto-< (2 → 6) -β-D-fructo- < isomalto < (2 → 1)-β-D-fructo- < gentio- < cello-oligosaccharides more specifically cyclo- (1 → 4)-α-D-gluco- < linear (1 → 4)-α-D-gluco- < (2 → 6)-β-D-fructo- < (1 → 6)-α-D-gluco- < (2 → 1)-β-D-fructo- <( 1 → 6)-β-D-gluco- < (1 → 4)-β-D-gluco-oligosaccharides. The particular behaviour of linear malto-oligosaccharides and the different behaviour of isomaltose makes the method still of great use for the assessment of starch-derived materials, including branch point residues. Specific size exclusion mechanisms were found not to operate for methyl ether or glycoside derivatives of monosaccharides although Bio-Gel P2 could be of limited use in methylation analysis and methyl glycoside separations.