Disaccharides

二糖
  • 文章类型: Journal Article
    木聚糖酶是木聚糖水解酶体系中最重要的水解酶,其主要功能是β-1,4-内切木聚糖酶,将木聚糖随机裂解为低聚木糖和木糖。木聚糖酶具有广泛的应用,但是对一些低温行业所需的冷适应酶的研究很少。据报道,糖苷水解酶家族8(GH8)木聚糖酶具有冷适应酶活性。在这项研究中,通过序列比对,从地热异常球菌中挖掘了木聚糖酶基因dgeoxyn。重组木聚糖酶DgeoXyn编码403个氨基酸,理论分子量为45.39kDa。结构分析表明,DgeoXyn具有典型的GH8木聚糖酶的(α/α)6桶折叠结构。同时,它具有严格的底物特异性,只对木聚糖有活性,其水解产物包括木二糖,木甲糖,木糖,二甲苯糖,和少量的木糖。DgeoXyn在70℃和pH6.0时最活跃。它在10、20和30℃下非常稳定,保留其最大酶活性的80%以上。DgeoXyn的酶活性在添加Mn2+后增加了10%,在添加Cu2+后降低了80%。dgeox的Km和Vmax分别为42mg/ml和20,000U/mg,分别,在温度为70℃和pH为6.0的条件下,使用10mg/ml山毛鱼木聚糖作为底物。本研究为低温木聚糖酶的开发和应用提供了理论依据。
    Xylanase is the most important hydrolase in the xylan hydrolase system, the main function of which is β-1,4-endo-xylanase, which randomly cleaves xylans to xylo-oligosaccharides and xylose. Xylanase has wide ranging of applications, but there remains little research on the cold-adapted enzymes required in some low-temperature industries. Glycoside hydrolase family 8 (GH8) xylanases have been reported to have cold-adapted enzyme activity. In this study, the xylanase gene dgeoxyn was excavated from Deinococcus geothermalis through sequence alignment. The recombinant xylanase DgeoXyn encodes 403 amino acids with a theoretical molecular weight of 45.39 kDa. Structural analysis showed that DgeoXyn has a (α/α)6-barrel fold structure typical of GH8 xylanase. At the same time, it has strict substrate specificity, is only active against xylan, and its hydrolysis products include xylobiose, xylotrinose, xytetranose, xylenanose, and a small amount of xylose. DgeoXyn is most active at 70 ℃ and pH 6.0. It is very stable at 10, 20, and 30 ℃, retaining more than 80% of its maximum enzyme activity. The enzyme activity of DgeoXyn increased by 10% after the addition of Mn2+ and decreased by 80% after the addition of Cu2+. The Km and Vmax of dgeox were 42 mg/ml and 20,000 U/mg, respectively, at a temperature of 70 ℃ and pH of 6.0 using 10 mg/ml beechwood xylan as the substrate. This research on DgeoXyn will provide a theoretical basis for the development and application of low-temperature xylanase.
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  • 文章类型: Journal Article
    硫酸乙酰肝素(HS)的生物活性与其分子量密切相关,硫酸化的程度和模式和均匀性。用于合成低分散性的均质糖链的现有方法涉及多个步骤并且需要逐步分离和纯化过程。这里,我们设计了一种用于包封糖基转移酶的介孔金属有机胶囊,并获得了能够酶催化聚合反应的微反应器,以制备低分散性的均相肝素,HS和肝素的前体。由于糖链延伸发生在微反应器的孔中,低分子量肝素可以通过空间限制催化合成。此外,糖基化副产物,尿苷二磷酸(UDP),可以与金属有机胶囊的暴露金属位点螯合,抑制反式切割以降低分子量分散性。这种微反应器提供了效率的优点,可重用性,并且消除了对逐步分离和纯化过程的需要。使用合成的肝素,我们进一步成功地制备了低分散性的均相6-O-硫酸化HS,其分子量约为6kDa,多分散指数(PDI)为1.032。值得注意的是,产生的HS表现出最小的抗凝血活性,其与成纤维细胞生长因子1的结合亲和力与低分子量肝素相当。
    The biological activities of heparan sulfate (HS) are intimately related to their molecular weights, degree and pattern of sulfation and homogeneity. The existing methods for synthesizing homogeneous sugar chains of low dispersity involve multiple steps and require stepwise isolation and purification processes. Here, we designed a mesoporous metal-organic capsule for the encapsulation of glycosyltransferase and obtained a microreactor capable of enzymatically catalyzing polymerization reactions to prepare homogeneous heparosan of low dispersity, the precursor of HS and heparin. Since the sugar chain extension occurs in the pores of the microreactor, low molecular weight heparosan can be synthesized through space-restricted catalysis. Moreover, the glycosylation co-product, uridine diphosphate (UDP), can be chelated with the exposed metal sites of the metal-organic capsule, which inhibits trans-cleavage to reduce the molecular weight dispersity. This microreactor offers the advantages of efficiency, reusability, and obviates the need for stepwise isolation and purification processes. Using the synthesized heparosan, we further successfully prepared homogeneous 6-O-sulfated HS of low dispersity with a molecular weight of approximately 6 kDa and a polydispersity index (PDI) of 1.032. Notably, the HS generated exhibited minimal anticoagulant activity, and its binding affinity to fibroblast growth factor 1 was comparable to that of low molecular weight heparins.
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  • 文章类型: Journal Article
    低聚半乳糖(GOS)是重要的益生元,其功能与其结构密切相关。然而,由于表征GOS中多种异构体的挑战,对结构-功能关系的全面概述仍然有限。本研究提出了一种将亲水相互作用液相色谱(HILIC)保留时间和串联质谱(MS/MS)片段模式相结合的策略,以通过HILIC-MS/MS分析区分GOS中二糖异构体的α/β键和键位置。结果表明,m/z203.0524与m/z365.1054的比值可以区分α/β键。而m/z347.0947与m/z365.1054,m/z245.0642与m/z365.1054和HILIC保留时间的比值可以区分(1→2),(1→3),(1→4)和(1→6)连杆。上述规则能够有效表征含GOS的食品样品中的二糖,包括奶粉,米粉,喝,酸奶。该方法可用于GOS的质量控制和未来对GOS结构特异性健康效应的研究。
    Galactooligosaccharides (GOS) are important prebiotics with function closely related to their structure. However, a comprehensive overview of the structure-function relationship is still limited due to the challenge in characterizing multiple isomers in GOS. This study presents a strategy of combining both hydrophilic interaction liquid chromatography (HILIC) retention time and tandem mass spectrometry (MS/MS) fragmentation pattern to distinguish α/β-linkages and linkage positions of disaccharide isomers in GOS through HILIC-MS/MS analysis. The results indicated that the ratio of m/z 203.0524 to m/z 365.1054 could distinguish α/β-linkages, while the ratios of m/z 347.0947 to m/z 365.1054, m/z 245.0642 to m/z 365.1054 and HILIC retention time could distinguish (1 → 2), (1 → 3), (1 → 4) and (1 → 6) linkages. The above rules enabled effective characterization of disaccharides in GOS-containing food samples, including milk powder, rice flour, drink, yogurt. This method can be used in the quality control of GOS and future research on the structure-specific health effects of GOS.
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  • 文章类型: Journal Article
    本研究旨在揭示HPLC指纹图谱的动力学,色度值,以及桑皮在炒制过程中的主要化学成分。建立了桑皮原料和加工产品的指纹图谱。莫伯兰甙A的含量,氧化白藜芦醇,kuwanonG,样品中的kuwanonH和样品的色度值被确定。此外,进行了指纹图谱的相似性评价和指纹图谱与色度值的相关性分析。结果表明,桑皮原料与加工品的指纹图谱具有较高的相似性,炒制过程中主要化学成分含量的总体变化相似。根据经验,当炒菜适中时,总色度值差|ΔE~*_(ab)|大于1.5。随着炒菜时间的延长,L~*和E~*_(ab)值不断减小,a~*值不断增加。指纹图谱与色度值的相关性分析结果表明,峰1(5-羟基麦芽酚),2(马伯罗塞德A),3,4,6,7,11(氧白藜芦醇),14、17(kuwanonG),和18(kuwanonH)与色度值显着相关。对含量较高的四种成分进行定量分析,结果表明,当炒菜过量时,四种成分的含量均有不同程度的下降。此外,5-羟基麦芽酚是在桑皮炒后生产的,指纹和色度值在炒制过程中呈规律性变化。色度可以包括在对MoriCortex的炒制过程的评估中,为规范炒桑皮质量提供参考。
    This study aims to reveal the dynamics of the HPLC fingerprint, chromaticity values, and main chemical components of Mori Cortex during the stir-frying process. The fingerprints of raw and processed products of Mori Cortex were established. The content of mulberroside A, oxyresveratrol, kuwanon G, and kuwanon H in the samples and the chromaticity values of the samples were determined. Furthermore, the similarity evaluation of fingerprints and the correlation analysis between fingerprints and chromaticity values were carried out. The results showed that the fingerprints of raw and processed products of Mori Cortex had high similarity, and the overall changes in the content of the main chemical components in the stir-frying process were similar. According to the experience, when the stir-frying is moderate, the total chromaticity value difference |ΔE~*_(ab)| is above 1.5. With the extension of stir-frying time, the L~* and E~*_(ab) values keep decreasing, and the a~* value keeps increasing. The results of the correlation analysis between fingerprints and chromaticity values showed that peaks 1(5-hydroxy maltol), 2(mulberroside A), 3, 4, 6, 7, 11(oxyresveratrol), 14, 17(kuwanon G), and 18(kuwanon H) had significant correlations with the chromaticity values. Quantitative analysis of the four components with higher content showed that the content of the four components decreased to varying degrees when the stir-frying was excessive. In addition, 5-hydroxy maltol was produced after stir-frying of Mori Cortex, and the fingerprint and chromaticity values showed regular changes during the stir-frying process. The chromaticity can be included in the evaluation of the stir-frying process of Mori Cortex, which provides a reference for standardizing the quality of stir-fried Mori Cortex.
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  • 文章类型: Journal Article
    背景:根据国家指南,低可发酵低聚糖的饮食,二糖,单糖,多元醇(FODMAP)是肠易激综合征(IBS)的二线治疗选择,可改善功能性肠道症状。在过去的十五年中,该领域已经发表了许多值得注意的结果。本研究旨在分析低FODMAP饮食研究的全球研究趋势和热点,为研究者提供全面的视角和方向。
    方法:使用WebofScienceCoreCollection(WoSCC)的科学引文索引扩展来识别与FODMAP饮食相关的低文章和评论。三个文献计量程序(CiteSpace,VOSviewer,ScimagoGraphic)用于分析和可视化年度出版物,作者,国家,机构,期刊,引文,和关键词。
    结果:总计,843份与低FODMAP饮食研究相关的文件由来自59个国家的1233个机构的3343名作者在227份期刊上发表。美国,这是参与国际合作最多的国家,年产量最大,增长最快。最有生产力的组织是莫纳什大学,最有成果的研究人员是吉布森公关。就已发表的文件数量而言,营养素排名第一。文章“低FODMAP饮食可减少肠易激综合征的症状”(HalmosEP,2014)获得了最多的共同引用。文献中经常出现的关键词主要涉及低FODMAP饮食的临床疗效评价和机制探索两个主要方面。术语“肠道微生物群”是迄今为止一直流行的突发关键词中最突出的关键词。
    结论:低FODMAP饮食的限制阶段在症状反应方面优于IBS的其他饮食疗法,但它对肠道双歧杆菌的丰度和饮食质量有负面影响。识别生物标志物以预测对低FODMAP饮食的反应是非常感兴趣的,并已成为当前的研究热点。
    BACKGROUND: According to national guidelines, a diet low in fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAP) is a second-line therapy option for irritable bowel syndrome (IBS) and improves functional intestinal symptoms. Numerous noteworthy results have been published in this field over the past fifteen years. This study aims to analyze the global research trend and hotspot of the low FODMAP diet research, and provide a comprehensive perspective and direction for researchers.
    METHODS: The Science Citation Index-Expanded of the Web of Science Core Collection (WoSCC) was used to identify low FODMAP diet-related articles and reviews. Three bibliometric programs (CiteSpace, VOSviewer, Scimago Graphic) were utilized to analyze and visualize the annual publications, authors, countries, institutions, journals, citations, and keywords.
    RESULTS: In total, 843 documents related to the low FODMAP diet research were published in 227 journals by 3,343 authors in 1,233 institutions from 59 countries. The United States, which was the most engaged nation in international collaboration, had the largest annual production and the fastest growth. The most productive organization was Monash University, and the most fruitful researcher was Gibson PR. Nutrients ranked first in terms of the number of published documents. The article \"A diet low in FODMAPs reduces symptoms of irritable bowel syndrome\" (Halmos EP, 2014) received the most co-citations. Keywords that appear frequently in the literature mainly involve two main aspects: the clinical efficacy evaluation and mechanism exploration of the low FODMAP diet. The term \"gut microbiota\" stands out as the most prominent keyword among the burst keywords that have remained prevalent till date.
    CONCLUSIONS: The restriction stage of the low FODMAP diet is superior to other dietary therapies for IBS in terms of symptom response, but it has a negative impact on the abundance of gut Bifidobacteria and diet quality. Identification of biomarkers to predict response to the low FODMAP diet is of great interest and has become the current research hotspot.
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  • 文章类型: Journal Article
    细菌感染是全球第二大死因,细菌病原体中抗生素抗性元素的进化和广泛分布加剧了威胁危机。碳水化合物参与细菌感染,耐药性和宿主免疫调节过程。来自碳水化合物或含有碳水化合物支架的许多抗菌剂有助于增加病原菌靶向,物理化学性质和可药用性。在论文中,根据从2014年至2024年的文献中收集的抗菌分子中含有的糖残基的类型和数量,抗菌活性,对作用机制和结构-活动关系进行了描述和总结,目的提供指导模板,以选择糖的类型和大小在寡糖为基础的抗微生物剂的设计,以对抗迫在眉睫的抗生素耐药性危机。
    Bacterial infections are the second leading cause of death worldwide, and the evolution and widespread distribution of antibiotic-resistance elements in bacterial pathogens exacerbate the threat crisis. Carbohydrates participate in bacterial infection, drug resistance and the process of host immune regulation. Numerous antimicrobials derived from carbohydrates or contained carbohydrate scaffolds that are conducive to an increase in pathogenic bacteria targeting, the physicochemical properties and druggability profiles. In the paper, according to the type and number of sugar residues contained in antimicrobial molecules collected from the literatures ranging from 2014 to 2024, the antimicrobial activities, action mechanisms and structure-activity relationships were delineated and summarized, for purpose to provide the guiding template to select the type and size of sugars in the design of oligosaccharide-based antimicrobials to fight the looming antibiotic resistance crisis.
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  • 文章类型: Journal Article
    具核梭杆菌,结直肠癌相关的微生物,可以触发或加速多种病理。我们报告了从F.nucleatumATCC23726O-抗原中首次合成含有六个氨基的缀合就绪二糖。稀有的2,3-二氨基-d-葡萄糖醛酸酰胺和2-乙酰氨基-4-氨基-d-岩藻糖由d-葡萄糖胺通过构型转化合成,亲核取代,C6氧化,和C6脱氧。保护基团和反应条件的明智选择能够选择性地安装N-乙酰基,N-丙酰基,N-甲酰基,和羧酰胺基。
    Fusobacterium nucleatum, a colorectal-cancer-associated oncomicrobe, can trigger or accelerate numerous pathologies. We report the first synthesis of a conjugation-ready disaccharide containing six amino groups from F. nucleatum ATCC 23726 O-antigen. Rare 2,3-diamido-d-glucuronic acid amide and 2-acetamido-4-amino-d-fucose were synthesized from d-glucosamine through configuration inversion, nucleophilic substitution, C6 oxidation, and C6 deoxygenation. A judicious choice of protecting groups and reaction conditions enabled the selective installation of N-acetyl, N-propanoyl, N-formyl, and carboxamido groups.
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  • 文章类型: Journal Article
    肝素是一种重要的抗凝血药物,和微生物肝素生物合成是一个潜在的替代动物来源的肝素生产。然而,有效使用肝素合成酶面临挑战,特别是活性硫酸乙酰肝素N-脱乙酰酶/N-磺基转移酶的微生物重组表达。这里,我们将单糖N-三氟乙酰氨基葡萄糖引入大肠杆菌K5以促进硫酸化修饰。蛋白质修复一站式服务聚焦的合理迭代位点特异性诱变(PROSS-FRISM)平台用于提高磺基转移酶效率,与野生型N-磺基转移酶相比,产生具有显著改善的稳定性(11.32倍)和活性(2.53倍)的工程化NST-M8酶。该方法可应用于工程化各种磺基转移酶。多酶级联反应能够从生物工程肝素生产活性肝素,显示抗FXa(246.09IU/mg)和抗FIIa(48.62IU/mg)活性。这项研究提供了克服肝素合成和修饰挑战的见解,使用基于细胞系统的半合成策略为无动物肝素的未来发展铺平了道路。
    Heparin is an important anticoagulant drug, and microbial heparin biosynthesis is a potential alternative to animal-derived heparin production. However, effectively using heparin synthesis enzymes faces challenges, especially with microbial recombinant expression of active heparan sulfate N-deacetylase/N-sulfotransferase. Here, we introduce the monosaccharide N-trifluoroacetylglucosamine into Escherichia coli K5 to facilitate sulfation modification. The Protein Repair One-Stop Service-Focused Rational Iterative Site-specific Mutagenesis (PROSS-FRISM) platform is used to enhance sulfotransferase efficiency, resulting in the engineered NST-M8 enzyme with significantly improved stability (11.32-fold) and activity (2.53-fold) compared to the wild-type N-sulfotransferase. This approach can be applied to engineering various sulfotransferases. The multienzyme cascade reaction enables the production of active heparin from bioengineered heparosan, demonstrating anti-FXa (246.09 IU/mg) and anti-FIIa (48.62 IU/mg) activities. This study offers insights into overcoming challenges in heparin synthesis and modification, paving the way for the future development of animal-free heparins using a cellular system-based semisynthetic strategy.
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  • 文章类型: Journal Article
    二果糖酸酐I(DFA-I)可以由菊粉生产,与DFA-I形成菊粉果糖转移酶(IFTase-I)。然而,菊粉通过DFA-I的代谢仍不清楚。为了澄清这条道路,合成了黄微杆菌DSM18909基因组中与该途径相关的几个酶基因,相应的酶被编码,纯化,并在体外进行了研究。菊粉通过IFTase-I分解为DFA-I后,通过DFA-I水解酶将DFA-I水解为氟尿糖。然后通过β-呋喃果糖苷酶水解红外二糖以形成果糖。最后,果糖通过果糖激酶进入糖酵解。β-呋喃果糖苷酶(MfFase1)清除副产物(蔗糖和低聚果糖),可能被果聚糖β-(2,1)-果糖苷酶/1-外水解酶和另一种呋喃果糖苷酶(MfFase2)部分水解。探索菊粉的DFA-I途径和体外研究良好的酶扩展了我们对菊粉能量提供方式的基本科学知识,从而为进一步的体内研究铺平了道路,并为将来菊粉和DFA-I的进一步营养研究提供了参考。
    Difructose anhydride I (DFA-I) can be produced from inulin, with DFA-I-forming inulin fructotransferase (IFTase-I). However, the metabolism of inulin through DFA-I remains unclear. To clarify this pathway, several genes of enzymes related to this pathway in the genome of Microbacterium flavum DSM 18909 were synthesized, and the corresponding enzymes were encoded, purified, and investigated in vitro. After inulin is decomposed to DFA-I by IFTase-I, DFA-I is hydrolyzed to inulobiose by DFA-I hydrolase. Inulobiose is then hydrolyzed by β-fructofuranosidase to form fructose. Finally, fructose enters glycolysis through fructokinase. A β-fructofuranosidase (MfFFase1) clears the byproducts (sucrose and fructo-oligosaccharides), which might be partially hydrolyzed by fructan β-(2,1)-fructosidase/1-exohydrolase and another fructofuranosidase (MfFFase2). Exploring the DFA-I pathway of inulin and well-studied enzymes in vitro extends our basic scientific knowledge of the energy-providing way of inulin, thereby paving the way for further investigations in vivo and offering a reference for further nutritional investigation of inulin and DFA-I in the future.
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  • 文章类型: Journal Article
    在这项研究中,我们研究了β-木糖苷酶基因的有害突变,木A(AkxylA),在鲁川曲霉中。KawachiiIFO4308通过构建来自A.luchuensisRIB2604(AlxylA)的AkxylA和木A的AkxylA破坏和互补菌株,它没有xylA的突变。只有AlxylA互补菌株在含有木聚糖的培养基中表现出显着的更高的生长和显着的β-木糖苷酶活性,伴随着XylA表达的增加。这导致大麦烧酒中的木二糖浓度较低,木糖浓度较高。这些发现表明xylA中的突变影响发酵过程中的木糖水平。因为xylA中的突变不仅在菌株IFO4308的基因组中被鉴定出来,而且在其他工业菌株的基因组中也被鉴定出来。Kawachii,这些发现增强了我们对影响发酵特性的遗传因素的理解。
    In this study, we investigated a deleterious mutation in the β-xylosidase gene, xylA (AkxylA), in Aspergillus luchuensis mut. kawachii IFO 4308 by constructing an AkxylA disruptant and complementation strains of AkxylA and xylA derived from A. luchuensis RIB2604 (AlxylA), which does not harbor the mutation in xylA. Only the AlxylA complementation strain exhibited significantly higher growth and substantial β-xylosidase activity in medium containing xylan, accompanied by an increase in XylA expression. This resulted in lower xylobiose and higher xylose concentrations in the mash of barley shochu. These findings suggest that the mutation in xylA affects xylose levels during the fermentation process. Because the mutation in xylA was identified not only in the genome of strain IFO 4308 but also the genomes of other industrial strains of A. luchuensis and A. luchuensis mut. kawachii, these findings enhance our understanding of the genetic factors that affect the fermentation characteristics.
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