Rhamnogalacturonan I

鼠李糖半乳糖醛酸 I
  • 文章类型: Journal Article
    人参Meyer多糖表现出多种生物学功能,如拮抗半乳糖凝集素-3介导的细胞粘附和迁移。半乳糖凝集素-8(Gal-8),具有接头连接的N端和C端碳水化合物识别结构域(CRD),对这些生物过程也至关重要,因此在各种病理障碍中起作用。然而,人参衍生的多糖在调节Gal-8功能中的作用仍不清楚。
    P.将人参来源的果胶进行色谱分离并酶消化以获得一系列多糖。生物层干涉法(BLI)量化了它们对Gal-8的结合亲和力,并通过血凝评估了它们对Gal-8的抑制作用,细胞迁移和T细胞凋亡。
    我们的人参衍生的果胶多糖主要由鼠李糖半乳糖醛酸-I(RG-I)和高半乳糖醛酸(HG)组成。BLI显示Gal-8结合主要位于RG-I及其β-1,4-半乳聚糖侧链,具有亚微摩尔KD值。N端和C端Gal-8CRD都结合RG-I,结合与Gal-8介导的功能相关。
    P.人参RG-I果胶β-1,4-半乳聚糖侧链对于结合Gal-8和拮抗其功能至关重要。这项研究增强了我们对半乳糖凝集素-糖相互作用的理解,可用于开发靶向Gal-8的药物的信息。
    UNASSIGNED: Panax ginseng Meyer polysaccharides exhibit various biological functions, like antagonizing galectin-3-mediated cell adhesion and migration. Galectin-8 (Gal-8), with its linker-joined N- and C-terminal carbohydrate recognition domains (CRDs), is also crucial to these biological processes, and thus plays a role in various pathological disorders. Yet the effect of ginseng-derived polysaccharides in modulating Gal-8 function has remained unclear.
    UNASSIGNED: P. ginseng-derived pectin was chromatographically isolated and enzymatically digested to obtain a series of polysaccharides. Biolayer Interferometry (BLI) quantified their binding affinity to Gal-8, and their inhibitory effects on Gal-8 was assessed by hemagglutination, cell migration and T-cell apoptosis.
    UNASSIGNED: Our ginseng-derived pectin polysaccharides consist mostly of rhamnogalacturonan-I (RG-I) and homogalacturonan (HG). BLI shows that Gal-8 binding rests primarily in RG-I and its β-1,4-galactan side chains, with sub-micromolar KD values. Both N- and C-terminal Gal-8 CRDs bind RG-I, with binding correlated with Gal-8-mediated function.
    UNASSIGNED: P. ginseng RG-I pectin β-1,4-galactan side chains are crucial to binding Gal-8 and antagonizing its function. This study enhances our understanding of galectin-sugar interactions, information that may be used in the development of pharmaceutical agents targeting Gal-8.
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  • 文章类型: Journal Article
    糖基化是一种通过将糖共价连接到蛋白质上来增强蛋白质功能特性的方法。本研究旨在制备三种偶联物(WP-HG,WP-SBP,和WP-RGI)通过干热方法研究了不同果胶结构对WP功能性质的影响,并表征了这些缀合物的性质和结构。研究结果表明,HG的糖基化程度(DG),SBP和RGI为13.13%±0.07%,分别为23.27%±0.3%和36.39%±0.3%,表明支链数量的增加促进了糖基化反应。通过FT-IR光谱技术鉴定缀合物的形成。SEM显示WP可以共价结合果胶,导致缀合物的更光滑和更致密的表面。圆二色性分析表明,糖基化反应改变了WP的二级结构,降低了α-螺旋含量。蛋白质空间构象的这种结构变化导致蛋白质表面疏水性降低。但果胶的加入进一步调节了蛋白质表面的亲水疏水比,从而提高WP的乳化性能。此外,糖基化可以提高乳液的稳定性,给它更小的液滴尺寸,更高的Zeta电位和更稳定的性能。一句话,本研究为不同结构的果胶在食品原料糖基化产品功能特性开发中的应用指明了方向。
    Glycosylation is a method that enhances the functional properties of proteins by covalently attaching sugars to them. This study aimed at preparing three conjugates (WP-HG, WP-SBP, and WP-RGI) by dry heating method to research the influence of different pectin structures on the functional properties of WP and characterize properties and structures of these conjugates. The research results manifested that the degree of glycosylation (DG) of HG, SBP and RGI were 13.13 % ± 0.07 %, 23.27 % ± 0.3 % and 36.39 % ± 0.3 % respectively, suggesting that the increase of the number of branch chains promoted the glycosylation reaction. The formation of the conjugate was identified by the FT-IR spectroscopy technique. And SEM showed that WP could covalently bind to pectin, resulting in a smoother and denser surface of the conjugates. The circular dichroism analysis exhibited that the glycosylation reaction altered the secondary structure of WP and decreased the α-Helix content. This structural change in the protein spatial conformation led to a decrease in the hydrophobicity of protein surface. But the addition of pectin further regulated the hydrophilic-hydrophobic ratio on the surface of the protein, thus improving the emulsification properties of WP. In addition, the glycosylation could improve the stability of the emulsion, giving it a smaller droplet size, higher Zeta-potential and more stable properties. In a word, this study pointed out the direction for the application of different pectin structures in the development of functional properties of glycosylation products in food ingredients.
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  • 文章类型: Journal Article
    这项研究是第一个从柑橘生理早熟果滴中提取和表征果胶的研究。酸水解法提取果胶的得率达到4.4%。柑橘生理早熟果滴果胶(CPDP)的甲氧基酯化度(DM)为15.27%,表明它是低甲氧基化果胶(LMP)。单糖组成和摩尔质量测试结果表明,CPDP是一种高度支化的大分子多糖(β:0.02,Mw:2.006×105g/mol),具有丰富的鼠李糖半乳糖醛酸I结构域(50.40%)和长的阿拉伯糖和半乳糖侧链(32.02%)。基于CPDP是LMP的事实,Ca2+用于诱导CPDP形成凝胶。质构和流变学测试表明,由于CPDP具有较低的DM和丰富的中性糖侧链,CPDP的凝胶强度和储能模量高于本文使用的商业柑橘果胶(CP)。扫描电子显微镜(SEM)结果显示CPDP具有稳定的凝胶网络结构。
    This study is the first to extract and characterize pectin from citrus physiological premature fruit drop. The extraction yield of pectin reached 4.4 % by acid hydrolysis method. The degree of methoxy-esterification (DM) of citrus physiological premature fruit drop pectin (CPDP) was 15.27 %, indicating it was low-methoxylated pectin (LMP). The monosaccharide composition and molar mass test results showed CPDP was a highly branched macromolecular polysaccharide (β: 0.02, Mw: 2.006 × 105 g/mol) with rich rhamnogalacturonan I domain (50.40 %) and long arabinose and galactose side chain (32.02 %). Based on the fact that CPDP is LMP, Ca2+ was used to induce CPDP to form gels. Textural and rheological tests showed that the gel strength and storage modulus of CPDP were higher than commercial citrus pectin (CP) used in this paper due to the lower DM and rich neutral sugar side chains of CPDP. Scanning electron microscope (SEM) results showed CPDP had stable gel network structure.
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  • 文章类型: Journal Article
    在这项研究中,通过人粪便分批发酵模型评估了高鼠李糖醛酸I果胶多糖(RGI)和自由基降解RGI(DRGI)的发酵特性,并对其结构特性进行了研究。因此,RGI的Mw从246.8下降到11.6kDa,树枝被戏剧性地折断了。发酵表明,RGI比DRGI降解更快,产生更多的乙酸盐和丙酸盐。两者都降低了厚壁菌/拟杆菌的比例,促进了拟杆菌的发展,双歧杆菌,和乳酸菌,给肠道生态系统带来好处。然而,RGI和DRGI中微生物群的组成和代谢途径不同。RGI的大多数优势细菌(例如[Eubacterium]_eligens_group)参与了碳水化合物的利用,导致更好的表现在糖脂代谢和能量代谢。这项工作阐明了大分子量在体外RGI型果胶多糖的肠道菌群调节作用中的作用。
    In this study, the fermentation characteristics of high rhamnogalacturonan I pectic polysaccharides (RGI) and free-radical degraded RGI (DRGI) were evaluated by a human fecal batch-fermentation model, and their structural properties were also investigated. As a result, the Mw of RGI decreased from 246.8 to 11.6 kDa, and the branches were broken dramatically. Fermentation showed that RGI degraded faster and produced more acetate and propionate than DRGI. Both of them reduced the Firmicutes/Bacteroidetes ratio and promoted the development of Bacteroides, Bifidobacterium, and Lactobacillus, bringing benefits to the gut ecosystem. However, the composition and metabolic pathways of the microbiota in RGI and DRGI were different. Most of the dominant bacteria of RGI (such as [Eubacterium]_eligens_group) participated in carbohydrate utilization, leading to better performance in glucolipid metabolism and energy metabolism. This work elucidated that large molecular weight matters in the gut microbiota modulatory effect of RGI-type pectic polysaccharides in vitro.
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  • 文章类型: Journal Article
    茶是一种受欢迎的饮料,具有悠久的安全和健康使用历史。茶多糖是从茶叶中提取的一种生物活性成分,近几十年来引起了越来越多的关注。在这篇文章中,通过热水提取和乙醇沉淀法从高古脑绿茶中分离出酸性多糖高古脑茶多糖(GPS)。通过二乙基氨基乙基(DEAE)SepharoseFastFlow柱和SephacrylS-400柱纯化后,获得了几种高半乳糖醛酸(HG)和鼠李糖半乳糖醛酸-I(RG-I)级分。通过甲基化和核磁共振(NMR)分析选择具有最高产率的级分GPS2b用于结构表征。发现GPS2b是HG型果胶多糖(甲酯化程度[DE],51.6%),具有低分子量(Mw,36.8kDa)。它主要由→4)-α-GalpA-(1→和→4)-α-GalpA-6-OMe-(1→。此外,在该部分中鉴定出次要的高度分支的RG-I结构域。研究茶多糖的结构特征可以为理解其结构-生物活性关系提供见解。
    Tea is a popular beverage with a long history of safe and healthy use. Tea polysaccharide is a bioactive component extracted from tea, which has attracted more and more attention in recent decades. In this article, an acidic polysaccharide Gougunao tea polysaccharide (GPS) was isolated from Gougunao green tea by hot water extraction and ethanol precipitation. After purification by a diethylaminoethyl (DEAE) Sepharose Fast Flow column and a Sephacryl S-400 column, several homogalacturonan (HG) and rhamnogalacturonan-I (RG-I) fractions were obtained. Fraction GPS2b with the highest yield was selected for structural characterization by methylation and nuclear magnetic resonance (NMR) analysis. GPS2b was found to be an HG-type pectic polysaccharide (degree of methyl esterification [DE], 51.6%) with low molecular weight (M w , 36.8 kDa). It was mainly composed of →4)-α-GalpA- (1→ and →4)-α-GalpA-6-OMe-(1→. In addition, a minor highly branched RG-I domain was identified in this fraction. The investigation of structural features of tea polysaccharides can provide insights to understand their structure-bioactivity relationship.
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  • 文章类型: Journal Article
    A low methyl-esterified pectin (33.2% methyl-esterification degree) was isolated from the tuber of Dioscorea opposita Thunb., which was an edible and medicinal material in China. This pectin (Mw of 1.3 × 104 g/mol) contained the ~59.1% homogalacturonan (HG) and ~38.1% highly branched rhamnogalacturonan I (RG-I) region with possible side chains embracing arabinogalactan II, arabinan or arabinogalactan I. The fragments including HG backbone consisting of → 4)-α-GalpA-(1 → and → 4)-α-GalpA-6-O-methyl-(1 → with molar ratio of ~2:1, and repeating unit of arabinogalactan II side chain composed of α-Araf-(1 → and → 3,6)-β-Galp-(1→, were speculated through methylation analysis and NMR spectra. However, the linkage pattern for RG-I backbone and side chains were indiscernible due to limited resolution of NMR spectra. Besides, the pectin adopted a flexible chain conformation in 0.1 M NaNO3 solution. These results provided a structural basis for study on polysaccharide from D. opposite, which was benefit for development of functional food of yam.
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  • 文章类型: Journal Article
    High pressure processing (HPP) has become a promising strategy for extracting bioactive constituents. In this study, the impact of HPP treatment at various pH values (2.0, 8.0, and 12.0) on the macromolecular, structural, antioxidant capacity, rheological characteristics and gel properties of citrus pectic polysaccharide was investigated. The results showed that pressure and pH significantly affected the yield and Rhamnogalacturonan I (RG-I) characterizations. The yields of high pressure extraction at pH 12 (28.13 %-33.95 %) were significantly higher than the yields at pH 2 (14.85 %-16.11 %) and pH 8 (8.75 %-9.65 %). The yield of HPP (500 MPa/10 min) assisted alkali extraction is more than 2 times of that of HPP assisted acid extraction. The RG-I structure ratio of HPP-alkali extraction pectic polysaccharide (74.51 %) was significantly higher than that of traditional pectin (41.83 %). The results showed that HPP assisted alkali is a potential pectic polysaccharide extraction technology.
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  • 文章类型: Journal Article
    A series of nucleotide sugar interconversion enzymes (NSEs) generate the activated sugar donors required for biosynthesis of cell wall matrix polysaccharides and glycoproteins. UDP-glucose 4-epimerases (UGEs) are NSEs that function in the interconversion of UDP-glucose (UDP-Glc) and UDP-galactose (UDP-Gal). The roles of UDP-glucose 4-epimerases in monocots remain unclear due to redundancy in the pathways. Here, we report a brittle plant (bp1) rice mutant that exhibits brittle leaves and culms at all growth stages. The mutant culms had reduced levels of rhamnogalacturonan I, homogalacturonan, and arabinogalactan proteins. Moreover, the mutant had altered contents of uronic acids, neutral noncellulosic monosaccharides, and cellulose. Map-based cloning demonstrated that OsBP1 encodes a UDP-glucose 4-epimerase (OsUGE2), a cytosolic protein. We also show that BP1 can form homo- and hetero-protein complexes with other UGE family members and with UDP-galactose transporters 2 (OsUGT2) and 3 (OsUGT3), which may facilitate the channeling of Gal to polysaccharides and proteoglycans. Our results demonstrate that BP1 participates in regulating the sugar composition and structure of rice cell walls.
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  • 文章类型: Journal Article
    Polysaccharides are considered to be the most important active substances in Goji. However, the structure of polysaccharides varies according to the extraction methods applied, and the solution used to prepare Goji polysaccharides (LBPs) were limited. Thus, it is important to clarify the connection between extraction methods and structure of Goji polysaccharide. In view of the complex composition of cell wall polysaccharides and the various forms of interaction, different extraction methods will release different parts of the cell wall. The present study compared the effects of different extraction methods, which have been used to prepare different types of plant cell wall polysaccharides based on various sources, on the structure of cell-wall polysaccharides from Goji, by the single separate use of hot water, hydrochloric acid (0.4%) and sodium hydroxide (0.6%), at both high and low temperatures. Meanwhile, in order to explore the limitations of single extraction, sequential extraction methods were applied. Structural analysis including monosaccharide analysis, GPC-MALLS, AFM and 1H-NMR suggested the persistence of more extensively branched rhamnogalacturonan I (RG-I) domains in the procedures involving low-temperature-alkali, while procedures prepared by high-temperature-acid contains more homogalacturonan (HG) regions and results in the removal of a substantial part of the side chain, specifically the arabinan. A kind of acidic heteropolysaccharide was obtained by hot water extraction. SEC-MALLS and AFM confirmed large-size polymers with branched morphologies in alkali-extracted polysaccharides. Our results provide new insight into the extraction of Goji polysaccharides, which differ from the hot water extraction used by traditional Chinese medicine.
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  • 文章类型: Journal Article
    O-Acetylated pectins are abundant in the primary cell wall of plants and growing evidence suggests they have important roles in plant cell growth and interaction with the environment. Despite their importance, genes required for O-acetylation of pectins are still largely unknown. In this study, we showed that TRICHOME BIREFRINGENCE LIKE 10 (AT3G06080) is involved in O-acetylation of pectins in Arabidopsis (Arabidopsis thaliana). The activity of the TBL10 promoter was strong in tissues where pectins are highly abundant (e.g. leaves). Two homozygous knock-out mutants of Arabidopsis, tbl10-1 and tbl10-2, were isolated and shown to exhibit reduced levels of wall-bound acetyl esters, equivalent of ~50% of the wild-type level in pectin-enriched fractions derived from leaves. Further fractionation revealed that the degree of acetylation of the pectin rhamnogalacturonan-I (RG-I) was reduced in the tbl10 mutant compared to the wild type, whereas the pectin homogalacturonan (HG) was unaffected. The degrees of acetylation in hemicelluloses (i.e. xyloglucan, xylan and mannan) were indistinguishable between the tbl10 mutants and the wild type. The mutant plants contained normal trichomes in leaves and exhibited a similar level of susceptibility to the phytopathogenic microorganisms Pseudomonas syringae pv. tomato DC3000 and Botrytis cinerea; while they displayed enhanced tolerance to drought. These results indicate that TBL10 is required for O-acetylation of RG-I, possibly as an acetyltransferase, and suggest that O-acetylated RG-I plays a role in abiotic stress responses in Arabidopsis.
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