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A HILIC-PDA(-MS) profiling approach for the analysis of (methyl esterified) unsaturated galacturonic acid oligosaccharides released from pectins and food by-products 用HILIC-PDA(-MS)分析果胶和食品副产品中释放的(甲基酯化)不饱和半乳糖醛酸低聚糖
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-12 DOI: 10.1016/j.carres.2026.109816
Rebekka Elke Schmidt, Mirko Bunzel
Functional and nutritional properties of pectins and pectic oligosaccharides (OS) depend on their structure. A major structural element of pectins are homogalacturonans, which consist of α-1,4-linked galacturonic acids (GalA). Homogalacturonans can be degraded to unsaturated (u)GalA-OS by pectin lyases. To analyze liberated uGalA-OS, a profiling approach based on hydrophilic interaction chromatography (HILIC) with photodiode array (PDA) and optional mass spectrometric (MS) detection was developed. Determination of molar relative response factors (RRF) of de-esterified uGalA-OS (degree of polymerization (DP) = 2–13) in relation to acarbose (internal standard) at 235 nm allows for (semi-)quantitative estimation of (methyl esterified) uGalA-OS and for application in other laboratories. Single quadrupole-MS enables to verify DP and to determine degree of methylation of uGalA-OS. Positions of methyl groups can be tentatively identified by orbitrap-MS. The developed profiling approach can be useful to analyze the specificity of a pectin lyase by analysis of uGalA-OS released from commercial pectin: The pectin lyase applied here appears to prefer a methyl group at cleavage subsite −1, but probably does not require a methyl group at subsite +1. In another application, various methyl esterified uGalA-OS (DP ≤ 12) were identified in enzymatically treated carrot pomace, a pectin-rich food by-product that can potentially be used to enhance nutritional properties of food products.
果胶和果胶寡糖(OS)的功能和营养特性取决于它们的结构。果胶的主要结构元素是半乳糖醛酸,它由α-1,4-链半乳糖醛酸(GalA)组成。高半乳糖醛酸可以通过果胶裂解酶降解为不饱和的(u)GalA-OS。为了分析游离uGalA-OS,建立了一种基于光电二极管阵列(PDA)和可选质谱(MS)检测的亲水性相互作用色谱(HILIC)分析方法。在235 nm下测定去酯化uGalA-OS(聚合度(DP) = 2-13)与阿卡波糖(内标)的摩尔相对响应因子(RRF),可以对(甲基酯化)uGalA-OS进行(半)定量估计,并在其他实验室中应用。单四极杆质谱可以验证DP并确定uGalA-OS的甲基化程度。甲基的位置可以用轨道谱法初步确定。所开发的分析方法可以通过分析商业果胶释放的uGalA-OS来分析果胶裂解酶的特异性:这里应用的果胶裂解酶似乎更倾向于在裂解亚位点- 1上有甲基,但可能不需要在亚位点+1上有甲基。在另一个应用中,在酶处理的胡萝卜渣中鉴定出各种甲基酯化的uGalA-OS (DP≤12),这是一种富含果胶的食品副产品,可以潜在地用于提高食品的营养特性。
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引用次数: 0
Chemical synthesis of the tetrasaccharide repeating unit of the O-antigen from halophilic Salinicola salarius HO-14 嗜盐盐菌HO-14 o抗原四糖重复单元的化学合成。
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-16 DOI: 10.1016/j.carres.2026.109813
Bijoy Rudra , Subrata Das , Balaram Mukhopadhyay
Total chemical synthesis of the tetrasaccharide repeating unit of the O-antigen from the halophilic bacterium Salinicola salarius HO-14 in the form of its 2-aminoethyl glycoside is reported. Partially protected monosaccharide synthons are prepared from the commercially available d-mannose and l-rhamnose through rational protecting group manipulations. Sequential stereoselective glycosylations through activation of the thioglycoside using NIS and TMSOTf gave the target tetrasaccharide. The crucial 1,2-cis rhamnosidic linkage is formed by the use of hydrogen bond assisted aglycon delivery. The conjugation ready target tetrasaccharide is attractive for further glycoconjugate formation.
报道了嗜盐细菌Salinicola salarius HO-14中o抗原的四糖重复单元的全化学合成,其形式为2-氨基乙基糖苷。从市售的d-甘露糖和l-鼠李糖中通过合理的保护基团操作制备部分保护的单糖合成物。通过使用NIS和TMSOTf激活巯基糖苷进行顺序立体选择性糖基化,得到目标四糖。关键的1,2-顺式鼠李糖键是通过氢键辅助糖苷传递形成的。偶联的目标四糖对进一步的糖偶联形成具有吸引力。
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引用次数: 0
Functionalization of β-glucans as a tool for enhancing biological properties: A scientometric approach β-葡聚糖功能化作为一种增强生物学特性的工具:一种科学计量方法
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-21 DOI: 10.1016/j.carres.2026.109836
Marcelo L.K. Marchioro , Gabrielly C.G. Xavier , Thaís F.M. Mota , Robert F.H. Dekker , Mário A.A. Cunha
β-glucans are polysaccharides with remarkable biological properties, including antioxidants, anticoagulants, antimicrobials, and immunomodulatory activities. However, their low solubility limits industrial applications, making chemical modification a key strategy to enhance solubility, biocompatibility, and biological performance. This scientometric study analyzed research on the chemical modification of β-glucans from 1986 to 2025, using data from Web of Science, Scopus, and PubMed. After refinement, 321 articles were evaluated through Excel, OriginPro 2025, and CiteSpace software. Results revealed a marked increase in publications and citations over the last decade, with China, Japan, the United States, and Brazil leading in productivity. Carbohydrate Polymers and the International Journal of Biological Macromolecules were the most influential journals in this field. Keyword analysis revealed three developmental stages: an initial phase centered on structural studies, an intermediate phase focused on biomedical applications (including antitumor and anticoagulant properties), and a recent phase emphasizing the antioxidant potential and integration of modified β-glucans in drug delivery and nanotechnology. Despite significant progress, challenges persist regarding the standardization of synthesis, structural characterization, and clinical validation. Strengthening regioselective synthesis, consistent analytical methods, and sustainable approaches is essential to consolidate modified β-glucans as promising biopolymers for pharmaceuticals, functional foods, and biomaterials.
β-葡聚糖是具有显著生物学特性的多糖,包括抗氧化剂、抗凝血剂、抗菌剂和免疫调节活性。然而,它们的低溶解度限制了工业应用,使化学改性成为提高溶解度,生物相容性和生物性能的关键策略。本科学计量学研究使用Web of Science、Scopus和PubMed的数据,分析了1986年至2025年间β-葡聚糖化学修饰的研究。精细化后,通过Excel、OriginPro 2025和CiteSpace软件对321篇文章进行评估。结果显示,在过去十年中,中国、日本、美国和巴西的出版物和引用量显著增加。《碳水化合物聚合物》和《国际生物大分子杂志》是该领域最具影响力的期刊。关键词分析揭示了三个发展阶段:以结构研究为中心的初始阶段,专注于生物医学应用(包括抗肿瘤和抗凝血特性)的中间阶段,以及强调抗氧化潜力和修饰β-葡聚糖在药物传递和纳米技术中的整合的最新阶段。尽管取得了重大进展,但在合成标准化、结构表征和临床验证方面仍然存在挑战。加强区域选择性合成、一致的分析方法和可持续的方法是巩固修饰β-葡聚糖作为有前途的生物聚合物用于药物、功能食品和生物材料的必要条件。
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引用次数: 0
A concise synthesis of the A and B blood group antigenic determinants A和B血型抗原决定因子的简明合成
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-13 DOI: 10.1016/j.carres.2026.109825
Fayliesha Spyker , Keith A. Stubbs
Novel and concise synthetic routes to the trisaccharide A and B blood group antigenic determinants have been devised. These trisaccharides, each containing a d-galactosyl-based and l-fucosyl moiety both α-linked to a central d-galactose unit, have previously been synthesised via a variety of methods, with most reporting poor yields. Developments in carbohydrate synthesis, especially the availability of highly α-selective galactosyl donors, provided the opportunity for novel and robust syntheses of these biologically relevant antigenic determinants to be explored. Herein are demonstrated efficient routes to these blood group antigenic determinants via initial establishment of the key α-1,3 linkage, followed by a late-stage incorporation of the l-fucosyl unit. These synthetic routes have the potential to be utilised in the preparation of similar compounds, with applications in ongoing research into the ABO blood group antigens and their influence on human health and disease states.
新的和简洁的合成路线,以三糖A和B血型抗原决定已设计。这些三糖,每个都含有一个基于d-半乳糖和l-聚焦基的部分,α-连接到中心的d-半乳糖单元,以前已经通过各种方法合成,大多数报道产量很低。碳水化合物合成的发展,特别是高α-选择性半乳糖供体的可用性,为探索这些生物相关抗原决定因子的新颖和强大的合成提供了机会。本文证明了通过关键α-1,3连锁的初始建立,随后晚期合并l- focusyl单元,有效途径到达这些血型抗原决定因子。这些合成途径有可能用于制备类似的化合物,并应用于正在进行的关于ABO血型抗原及其对人类健康和疾病状态影响的研究。
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引用次数: 0
Investigation of monosaccharide colorimetric efficiency by phenol-sulfuric acid method and optimization of quantitative models for polysaccharides 苯酚-硫酸法测定单糖比色效率及多糖定量模型优化
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-20 DOI: 10.1016/j.carres.2026.109835
Anbang Li , Chang Liu , Ronge Xing , Song Liu , Kecheng Li
Phenol-sulfuric acid method, a classical approach for total sugar quantification, suffers from substantial variation in the colorimetric efficiency among different monosaccharides, which may result in significant measurement bias. In this study, we investigated the colorimetric efficiency of 14 monosaccharides across three different systems and found that the colorimetric efficiency followed the pattern: neutral > acidic > amino monosaccharides. Notably, the colorimetric efficiency of mannose was over 800-fold higher than that of glucosamine. Principal coordinate analysis (PCoA) revealed that functional groups (carboxyl, amino) were the major determinants of colorimetric efficiency. Based on these findings, we assessed three quantitative models for five polysaccharides. The traditional model was suitable for homogeneous glucans (e.g., starch, recovery ∼101.0 %). The dominant monosaccharide model was effective for neutral polysaccharides with a single monosaccharide composition (e.g., mannan, recovery ∼100.5 % in the optimized system). In contrast, the weighted model, which integrates molar ratios and individual efficiencies, failed to improve accuracy for complex heterogeneous acidic polysaccharides (e.g., fucoidans, recovery <40.0 %). Furthermore, amino groups in polysaccharides (e.g., chitosan) strongly inhibited the reaction, precluding accurate quantification. This study provides a theoretical basis for the precise application of the phenol-sulfuric acid method and offers methodological guidance for the standardization of polysaccharide analysis.
苯酚-硫酸法是一种经典的总糖定量方法,不同单糖的比色效率存在较大差异,可能导致显著的测量偏差。在本研究中,我们研究了14种单糖在三种不同体系中的比色效率,发现比色效率遵循中性>;酸性>;氨基单糖的模式。甘露糖比葡萄糖胺的比色效率高800倍以上。主配位分析(PCoA)表明,官能团(羧基、氨基)是影响比色效率的主要因素。基于这些发现,我们评估了五种多糖的三种定量模型。传统模型适用于均质葡聚糖(如淀粉,回收率为~ 101.0%)。优势单糖模型对单一单糖组成的中性多糖有效(例如甘露聚糖,在优化体系中回收率为~ 100.5%)。相比之下,整合摩尔比和个体效率的加权模型未能提高复杂的非均相酸性多糖(例如岩藻多糖,回收率为40.0%)的准确性。此外,多糖(如壳聚糖)中的氨基强烈抑制了该反应,妨碍了准确的定量。本研究为苯酚-硫酸法的精确应用提供了理论依据,为多糖分析的规范化提供了方法学指导。
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引用次数: 0
Oligosaccharides from Scorzonera yildirimlii and S. zorkunensis, and their potential antimicrobial, enzyme inhibition, cytotoxic effect, and in silico study 山葵和山葵的低聚糖,及其潜在的抗菌、酶抑制、细胞毒作用,和硅的研究
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-16 DOI: 10.1016/j.carres.2026.109833
Nurettin Yaylı , Büşra Korkmaz , Gozde Bozdal , Büşra Şahin , Şeyma Arıcı , Yasemin Altun Ali , Sermet Yıldırmış , Gökçe Seyhan , Ahmet Buğra Aksel , Kamil Coşkunçelebi , Serdar Makbul , Arif Bozdeveci
<div><div>In this study, the phytochemical investigation of <em>Scorzonera yildirimlii</em> and <em>S. zorkunensis</em> led to the isolation and identification of eight new oligosaccharide constituents (<strong>1</strong>–<strong>8</strong>). Their structures were elucidated through comprehensive spectroscopic analysis (NMR, FT-IR, and mass spectrometry) and comparison with literature data. The structure of isolated compounds were <em>O</em>-<em>α</em>-D-Glc<em>p-</em>(1 → 2)-<em>O</em>-<em>β</em>-D-Fru<em>f</em>-(3 → 1)-<em>O</em>-<em>myo</em>-inositol (<strong>1</strong>), <em>O</em>-<em>α</em>-D-Glc<em>p-</em>(1 → 2)-<em>O</em>-<em>β</em>-D-Fru<em>f</em>-(3 → 1)-<em>O</em>-<em>epi</em>-inositol (<strong>2</strong>), <em>O</em>-<em>β</em>-D-Fru<em>f</em>-(2 → 6)-<em>O</em>-<em>α</em>-D-Glc<em>p</em>-(1 → 1)-<em>O</em>-<em>β</em>-D-Glc<em>p</em> (<strong>3</strong>), <em>O</em>-<em>β</em>-D-Fru<em>f</em>-(2 → 6)-<em>O</em>-α-D-Glc<em>p</em>-(1 → 1)-<em>O</em>-<em>β</em>-D-Gal<em>p</em> (<strong>4</strong>), <em>O</em>-<em>β</em>-D-Fru<em>f</em>-(2 → 6)-<em>O</em>-<em>β</em>-D-Fru<em>f</em>-(2 → 6)-<em>O</em>-<em>α</em>-D-Glc<em>p</em>-(1 → 1)-<em>O</em>-<em>β</em>-D-Glc<em>p</em> (<strong>5</strong>), <em>O</em>-<em>β</em>-D-Fru<em>f</em>-(2 → 6)-[<em>O</em>-<em>β</em>-D-Glc<em>p-</em>(1 → 4)]-<em>O</em>-<em>α</em>-D-Glc<em>p</em>-(1 → 1)-<em>O</em>-<em>β</em>-D-Glc<em>p</em> (<strong>6</strong>), <em>O</em>-<em>β</em>-D-Fru<em>f</em>-(2 → 6)-[<em>O</em>-<em>β</em>-D-Glc<em>p</em>-(1 → 4)]-3-OCH<sub>3</sub>-<em>O</em>-<em>α</em>-D-Glc<em>p</em>-(1 → 1)-<em>O</em>-<em>β</em>-D-Glc<em>p</em> (<strong>7</strong>), and <em>O</em>-<em>β</em>-D-Fru<em>f</em>-(2 → 6)-[<em>O</em>-<em>β</em>-D-Glc<em>p</em>-(1 → 4)]-<em>O</em>-<em>α</em>-D-Glc<em>p</em>-(1 → 4)-<em>O</em>-<em>α</em>-D-Glc<em>p-</em>(1 → 2)-O-<em>β</em>-D-Fru<em>f</em> (<strong>8</strong>). The antimicrobial activities of all the isolated compounds (<strong>1</strong>–<strong>8</strong>) were investigated against six microorganisms. Compounds <strong>2</strong> and <strong>3</strong> were effective against <em>Mycobacterium smegmatis</em> at minimum inhibitory concentrations (MICs) of 3.13 μg/mL and 3.44 μg/mL, respectively, compared with streptomycin (MIC = 4.0 μg/mL). The enzyme-inhibitory properties of the isolated compounds (<strong>1</strong>–<strong>8</strong>) were evaluated against tyrosinase and <em>α</em>-glucosidase. Among the tested compounds, <strong>4</strong> (39.78 ± 10.29 %) exhibited inhibitory activity against tyrosinase, while compound <strong>2</strong> (68.04 ± 4.68 %) exhibited inhibitory activity against α-glucosidase. <em>In silico</em> studies of <strong>1</strong>–<strong>8</strong> showed the best predicted binding affinities for the Trehalose-LpqY complex in the range of −11.088 kcal/mol to −14.297 kcal/mol, compared to streptomycin (−6.638 kcal/mol). Compounds <strong>1</strong>–<strong>8</strong> exhibited almost no cytotoxic activity against L-929 cells, fu
本研究通过对Scorzonera yildirimlii和S. zorkunensis的植物化学研究,分离鉴定了8个新的低聚糖成分(1-8)。通过综合光谱分析(NMR、FT-IR和质谱)和文献资料对比,对其结构进行了鉴定。孤立的结构化合物O -α-D-Glcp -(1→2)- O -β-D-Fruf -(3→1)-O-myo-inositol (1), O -α-D-Glcp -(1→2)- O -β-D-Fruf -(3→1)-O-epi-inositol (2), O -β-D-Fruf -(2→6)- O -α-D-Glcp -(1→1)- O -β-D-Glcp (3), O -β-D-Fruf -(2→6)- O -α-D-Glcp -(1→1)- O -β-D-Galp (4), O -β-D-Fruf -(2→6)- O -β-D-Fruf -(2→6)- O -α-D-Glcp -(1→1)- O -β-D-Glcp (5), O -β-D-Fruf -(2→6)[O -β-D-Glcp -(1→4)- O -α-D-Glcp -(1→1)- O -β-D-Glcp (6), O -β-D-Fruf -(2→6)[O -β-D-Glcp -(1→4)]3-och3-o -α-D-Glcp -(1→1)- O -β-D-Glcp (7),和O -β-D-Fruf -(2→6)[O -β-D-Glcp -(1→4)- O -α-D-Glcp -(1→4)- O -α-D-Glcp -(1→2)- O -β-D-Fruf(8)。研究了分离得到的化合物(1 ~ 8)对6种微生物的抑菌活性。化合物2和3对耻垢分枝杆菌的最低抑制浓度(MIC)分别为3.13 μg/mL和3.44 μg/mL,而链霉素的最低抑制浓度为4.0 μg/mL。分离得到的化合物(1-8)对酪氨酸酶和α-葡萄糖苷酶的酶抑制性能进行了评价。化合物4(39.78±10.29%)对酪氨酸酶具有抑制活性,化合物2(68.04±4.68%)对α-葡萄糖苷酶具有抑制活性。硅实验结果表明,海藻糖- lpqy复合物的结合亲和力在- 11.088 kcal/mol至- 14.297 kcal/mol之间,而链霉素的结合亲和力为- 6.638 kcal/mol。化合物1-8对L-929细胞几乎没有细胞毒活性,进一步支持其治疗用途的潜力。
{"title":"Oligosaccharides from Scorzonera yildirimlii and S. zorkunensis, and their potential antimicrobial, enzyme inhibition, cytotoxic effect, and in silico study","authors":"Nurettin Yaylı ,&nbsp;Büşra Korkmaz ,&nbsp;Gozde Bozdal ,&nbsp;Büşra Şahin ,&nbsp;Şeyma Arıcı ,&nbsp;Yasemin Altun Ali ,&nbsp;Sermet Yıldırmış ,&nbsp;Gökçe Seyhan ,&nbsp;Ahmet Buğra Aksel ,&nbsp;Kamil Coşkunçelebi ,&nbsp;Serdar Makbul ,&nbsp;Arif Bozdeveci","doi":"10.1016/j.carres.2026.109833","DOIUrl":"10.1016/j.carres.2026.109833","url":null,"abstract":"&lt;div&gt;&lt;div&gt;In this study, the phytochemical investigation of &lt;em&gt;Scorzonera yildirimlii&lt;/em&gt; and &lt;em&gt;S. zorkunensis&lt;/em&gt; led to the isolation and identification of eight new oligosaccharide constituents (&lt;strong&gt;1&lt;/strong&gt;–&lt;strong&gt;8&lt;/strong&gt;). Their structures were elucidated through comprehensive spectroscopic analysis (NMR, FT-IR, and mass spectrometry) and comparison with literature data. The structure of isolated compounds were &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p-&lt;/em&gt;(1 → 2)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(3 → 1)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;myo&lt;/em&gt;-inositol (&lt;strong&gt;1&lt;/strong&gt;), &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p-&lt;/em&gt;(1 → 2)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(3 → 1)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;epi&lt;/em&gt;-inositol (&lt;strong&gt;2&lt;/strong&gt;), &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(2 → 6)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 1)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt; (&lt;strong&gt;3&lt;/strong&gt;), &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(2 → 6)-&lt;em&gt;O&lt;/em&gt;-α-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 1)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Gal&lt;em&gt;p&lt;/em&gt; (&lt;strong&gt;4&lt;/strong&gt;), &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(2 → 6)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(2 → 6)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 1)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt; (&lt;strong&gt;5&lt;/strong&gt;), &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(2 → 6)-[&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Glc&lt;em&gt;p-&lt;/em&gt;(1 → 4)]-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 1)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt; (&lt;strong&gt;6&lt;/strong&gt;), &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(2 → 6)-[&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 4)]-3-OCH&lt;sub&gt;3&lt;/sub&gt;-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 1)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt; (&lt;strong&gt;7&lt;/strong&gt;), and &lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt;-(2 → 6)-[&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;β&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 4)]-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p&lt;/em&gt;-(1 → 4)-&lt;em&gt;O&lt;/em&gt;-&lt;em&gt;α&lt;/em&gt;-D-Glc&lt;em&gt;p-&lt;/em&gt;(1 → 2)-O-&lt;em&gt;β&lt;/em&gt;-D-Fru&lt;em&gt;f&lt;/em&gt; (&lt;strong&gt;8&lt;/strong&gt;). The antimicrobial activities of all the isolated compounds (&lt;strong&gt;1&lt;/strong&gt;–&lt;strong&gt;8&lt;/strong&gt;) were investigated against six microorganisms. Compounds &lt;strong&gt;2&lt;/strong&gt; and &lt;strong&gt;3&lt;/strong&gt; were effective against &lt;em&gt;Mycobacterium smegmatis&lt;/em&gt; at minimum inhibitory concentrations (MICs) of 3.13 μg/mL and 3.44 μg/mL, respectively, compared with streptomycin (MIC = 4.0 μg/mL). The enzyme-inhibitory properties of the isolated compounds (&lt;strong&gt;1&lt;/strong&gt;–&lt;strong&gt;8&lt;/strong&gt;) were evaluated against tyrosinase and &lt;em&gt;α&lt;/em&gt;-glucosidase. Among the tested compounds, &lt;strong&gt;4&lt;/strong&gt; (39.78 ± 10.29 %) exhibited inhibitory activity against tyrosinase, while compound &lt;strong&gt;2&lt;/strong&gt; (68.04 ± 4.68 %) exhibited inhibitory activity against α-glucosidase. &lt;em&gt;In silico&lt;/em&gt; studies of &lt;strong&gt;1&lt;/strong&gt;–&lt;strong&gt;8&lt;/strong&gt; showed the best predicted binding affinities for the Trehalose-LpqY complex in the range of −11.088 kcal/mol to −14.297 kcal/mol, compared to streptomycin (−6.638 kcal/mol). Compounds &lt;strong&gt;1&lt;/strong&gt;–&lt;strong&gt;8&lt;/strong&gt; exhibited almost no cytotoxic activity against L-929 cells, fu","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"562 ","pages":"Article 109833"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146036014","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Dissecting the glycoprofile of the human cytomegalovirus protein gB by comprehensive mass spectrometry analysis 用综合质谱法分析人巨细胞病毒蛋白gB的糖谱
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-26 DOI: 10.1016/j.carres.2026.109838
Antonio Lembo , Gian Luca Sardone , Michela Aurilia , Antonio Molinaro , Francesco Berti , Cristina De Castro , Massimiliano Biagini
Glycosylation is the most critical post-translational modification occurring on proteins and its specific pattern depends both on the cells used for their production, such as mammalian cells, insect cells and yeast, and the expression conditions (e.g., mannosidase inhibitors, gene deletion). The aim of this study is to characterize the glycans of recombinant glycoprotein B (gB) from Human Cytomegalovirus (HCMV), expressed in CHO cells, by using different hyphenated mass spectrometry techniques in order to enrich the knowledge about glycosylation pattern changes in the protein production process and to develop versatile analytical platforms for the rapid analysis of protein glycosylation. The settled workflows focus on two main classes of analytes originating from the glycoprotein studied: glycopeptides and glycans. The analysis of the first category of molecules relies on a bottom-up proteomic approach in which the glycosites and the correspondent glycan heterogeneity and occupancy have been investigated by using high resolution tandem mass spectrometry; the analysis of the second class of molecules occurs by glycan shaving followed by fluorescence-liquid chromatography and mass-spectrometry characterization. These analyzes have been performed on the same glycoprotein expressed in a different culture media to demonstrate the capability of these analytical methods to monitor in detail oligosaccharide pattern changes. Finally, the impact of changes in glycosylation pattern on antigen immunogenicity has been assessed, showing that the presence of only high mannose oligosaccharides elicits higher titer of neutralizing antibodies.
糖基化是发生在蛋白质上的最关键的翻译后修饰,其具体模式取决于用于其生产的细胞,如哺乳动物细胞、昆虫细胞和酵母,以及表达条件(如甘露糖苷酶抑制剂、基因缺失)。本研究的目的是利用不同的联用质谱技术对CHO细胞中表达的人巨细胞病毒(HCMV)重组糖蛋白B (gB)的聚糖进行表征,以丰富蛋白质生产过程中糖基化模式变化的知识,并为蛋白质糖基化的快速分析开发通用的分析平台。确定的工作流程集中在两类主要的分析物上,这些分析物来源于所研究的糖蛋白:糖肽和聚糖。第一类分子的分析依赖于自下而上的蛋白质组学方法,其中使用高分辨率串联质谱法研究了糖基和相应的糖基异质性和占用;第二类分子的分析是通过聚糖剃除,然后进行荧光-液相色谱和质谱表征。这些分析是在不同培养基中表达的相同糖蛋白上进行的,以证明这些分析方法能够详细监测低聚糖模式的变化。最后,糖基化模式的变化对抗原免疫原性的影响已被评估,表明只有高甘露糖低聚糖的存在引起更高滴度的中和抗体。
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引用次数: 0
Characterization of a new glycosaminoglycan-like exopolysaccharide produced by the marine bacterium Alteromonas macleodii Mo169 海洋细菌Alteromonas macleodii Mo169产生的一种新的糖胺聚糖样胞外多糖的性质。
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-30 DOI: 10.1016/j.carres.2026.109847
Silvia Fanina , Patrícia Concórdio-Reis , Xavier Moppert , Filomena Freitas , Angela Casillo , Maria Michela Corsaro
Marine microorganisms produce extracellular polysaccharides with unique features, such as the presence of unusual monosaccharides, adhesiveness to biotic and abiotic surfaces, and close similarity to glycosaminoglycans (GAG), that are of considerable interest for biotechnological applications. In the present study, an extracellular microbial polysaccharide (EPS) has been isolated and structurally characterized. The EPS producer is the Gram-negative bacterium Alteromonas macleodii Mo169, previously isolated from a marine environment in French Polynesia. The bacterium has been cultivated using glycerol as the sole carbon source. Interestingly, the secreted EPS was composed of 2-acetamido-2-deoxy-guluronic acid (GulNA) and 2-acetamido-2-deoxy-glucose (GlcN), two amino sugars not commonly found in the Alteromonas genus. The purified EPS, with a molecular weight of 0.30 ± 0.01 MDa, consisted of the following repeating unit: →4)-α-L-GulNAc3OAcA-(1 → 3)-β-D-GlcNAc-(1 → .
海洋微生物产生的胞外多糖具有独特的特征,如不寻常的单糖的存在,对生物和非生物表面的粘附性,以及与糖胺聚糖(GAG)的密切相似性,这对生物技术应用具有相当大的兴趣。本研究分离了一种胞外微生物多糖(EPS),并对其进行了结构表征。EPS的产生者是革兰氏阴性菌变单胞菌Mo169,以前从法属波利尼西亚的海洋环境中分离出来。这种细菌是用甘油作为唯一的碳源培养出来的。有趣的是,分泌的EPS由2-乙酰氨基-2-脱氧古鲁醛酸(GulNA)和2-乙酰氨基-2-脱氧葡萄糖(GlcN)组成,这两种氨基糖在Alteromonas属中并不常见。纯化的EPS分子量为0.30±0.01 MDa,由以下重复单元组成:→4)-α- l - gulnac3oaca -(1→3)-β- d - glcnac -(1→)。
{"title":"Characterization of a new glycosaminoglycan-like exopolysaccharide produced by the marine bacterium Alteromonas macleodii Mo169","authors":"Silvia Fanina ,&nbsp;Patrícia Concórdio-Reis ,&nbsp;Xavier Moppert ,&nbsp;Filomena Freitas ,&nbsp;Angela Casillo ,&nbsp;Maria Michela Corsaro","doi":"10.1016/j.carres.2026.109847","DOIUrl":"10.1016/j.carres.2026.109847","url":null,"abstract":"<div><div>Marine microorganisms produce extracellular polysaccharides with unique features, such as the presence of unusual monosaccharides, adhesiveness to biotic and abiotic surfaces, and close similarity to glycosaminoglycans (GAG), that are of considerable interest for biotechnological applications. In the present study, an extracellular microbial polysaccharide (EPS) has been isolated and structurally characterized. The EPS producer is the Gram-negative bacterium <em>Alteromonas macleodii</em> Mo169, previously isolated from a marine environment in French Polynesia. The bacterium has been cultivated using glycerol as the sole carbon source. Interestingly, the secreted EPS was composed of 2-acetamido-2-deoxy-guluronic acid (GulNA) and 2-acetamido-2-deoxy-glucose (GlcN), two amino sugars not commonly found in the <em>Alteromonas</em> genus. The purified EPS, with a molecular weight of 0.30 ± 0.01 MDa, consisted of the following repeating unit: →4)-α-L-GulNAc3OAcA-(1 → 3)-β-D-GlcNAc-(1 → .</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"562 ","pages":"Article 109847"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146099880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Heterologous expression, and characterization of chitinase from Kitasatospora setae and its efficient conversion of chitin polymer into chitobiose Kitasatospora setae几丁质酶的异源表达、鉴定及几丁质聚合物转化为几丁糖的效率
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-12 DOI: 10.1016/j.carres.2026.109815
Jiacheng Zhang , Haorui Xu , Xingxing Fang , Chong Zhang
Using chitinase to convert chitin into oligosaccharides is greener compared to chemical methods. However, the low activity and low yield of existing chitinase enzymes limit their use. Here we reported a chitinase gene of Kitasatospora setae extracted from Uniprot, the chitinase was named KsChi. KsChi contains a signal peptide, a catalytic domain, and a chitin-binding domain. Insertion of the KsChi gene fragment into the pET-28a vector followed by transformation into Escherichia coli BL21(DE3) cells resulted in active expression. KsChi was optimally active at 60 °C in 50 mM citric acid buffer pH 5.0. KsChi exhibited the highest activity towards colloidal chitin, it showed a Kcat as 19.75 s−1 and overall catalytic efficiency (Kcat/KM) as 19.17 mL mg−1 s−1. KsChi can hydrolyze colloidal chitin to produce chitin oligosaccharides and has good stability under optimal reaction conditions. We expressed KsChi in a 5-L bioreactor, with a yield of up to 500 mg L−1, which is currently the highest level, the total activity is approximately 11 times higher than those reported in other studies. Our results indicate that KsChi is an exochitinase, and its main hydrolysis product is N, N′-diacetylchitobiose ((GlcNAc)2), which has a low KM value and high activity, thus having a wide range of application prospects.
与化学方法相比,利用几丁质酶将几丁质转化为低聚糖更为环保。然而,现有几丁质酶的低活性和低产量限制了它们的使用。本文报道了从Uniprot中提取的Kitasatospora setae几丁质酶基因,并将其命名为KsChi。KsChi含有一个信号肽、一个催化结构域和一个几丁质结合结构域。将KsChi基因片段插入pET-28a载体,然后转化到大肠杆菌BL21(DE3)细胞中,产生活性表达。KsChi在60℃、50 mM柠檬酸缓冲液pH 5.0条件下活性最佳。KsChi对胶体甲壳素的催化活性最高,Kcat为19.75 s−1,总催化效率(Kcat/KM)为19.17 mL mg−1 s−1。KsChi可水解胶体几丁质制备几丁质低聚糖,并在最佳反应条件下具有良好的稳定性。我们在5-L的生物反应器中表达了KsChi,产率高达500 mg L−1,这是目前最高的水平,总活性大约是其他研究报道的11倍。我们的研究结果表明,KsChi是一种外几丁质酶,其主要水解产物为N, N ' -二乙酰壳聚糖((GlcNAc)2), KM值低,活性高,具有广泛的应用前景。
{"title":"Heterologous expression, and characterization of chitinase from Kitasatospora setae and its efficient conversion of chitin polymer into chitobiose","authors":"Jiacheng Zhang ,&nbsp;Haorui Xu ,&nbsp;Xingxing Fang ,&nbsp;Chong Zhang","doi":"10.1016/j.carres.2026.109815","DOIUrl":"10.1016/j.carres.2026.109815","url":null,"abstract":"<div><div>Using chitinase to convert chitin into oligosaccharides is greener compared to chemical methods. However, the low activity and low yield of existing chitinase enzymes limit their use. Here we reported a chitinase gene of <em>Kitasatospora setae</em> extracted from Uniprot, the chitinase was named KsChi. KsChi contains a signal peptide, a catalytic domain, and a chitin-binding domain. Insertion of the KsChi gene fragment into the pET-28a vector followed by transformation into <em>Escherichia coli</em> BL21(DE3) cells resulted in active expression. KsChi was optimally active at 60 °C in 50 mM citric acid buffer pH 5.0. KsChi exhibited the highest activity towards colloidal chitin, it showed a <em>K</em><sub>cat</sub> as 19.75 s<sup>−1</sup> and overall catalytic efficiency (<em>K</em><sub>cat</sub>/<em>K</em><sub>M</sub>) as 19.17 mL mg<sup>−1</sup> s<sup>−1</sup>. KsChi can hydrolyze colloidal chitin to produce chitin oligosaccharides and has good stability under optimal reaction conditions. We expressed KsChi in a 5-L bioreactor, with a yield of up to 500 mg L<sup>−1</sup>, which is currently the highest level, the total activity is approximately 11 times higher than those reported in other studies. Our results indicate that KsChi is an exochitinase, and its main hydrolysis product is <em>N</em>, <em>N′</em>-diacetylchitobiose ((GlcNAc)<sub>2</sub>), which has a low <em>K</em><sub>M</sub> value and high activity, thus having a wide range of application prospects.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"562 ","pages":"Article 109815"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145950154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
α-lipoic acid and L-serine conjugated chitosan as a stimuli-responsive drug delivery platform α-硫辛酸-l -丝氨酸偶联壳聚糖作为刺激反应性给药平台的研究
IF 2.5 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2026-04-01 Epub Date: 2026-01-21 DOI: 10.1016/j.carres.2026.109837
Özlem Kaplan , Kevser Bal , Sibel Küçükertuğrul Çeli̇k , Nazan Gökşen Tosun , Mehmet Koray Gök
In this study, a novel chitosan-based nanocarrier system was developed through dual modification with α-lipoic acid and l-serine (ChiLipo–Ser) to enhance its physicochemical and biological properties for drug delivery applications. FTIR and 1H NMR spectroscopy verified the successful conjugation of functional groups onto the chitosan backbone, while GPC analysis confirmed an increase in molecular weight consistent with the modification. Compared with native chitosan, the ChiLipo–Ser exhibited improved solubility at physiological pH and enhanced buffering capacity. Nanoparticles prepared via ionotropic gelation demonstrated favorable characteristics, including an average particle size of ∼150 nm, low polydispersity, and a positive surface charge. Redox sensitivity was evidenced by DLS analysis, which revealed particle destabilization in the presence of 10–20 mM DTT, resulting from disulfide bond cleavage. Importantly, ChiLipo–Ser modification significantly enhanced the encapsulation efficiency of curcumin compared to unmodified chitosan, while the resulting formulations also exhibited effective release behavior at pH 5, mimicking the tumor intracellular environment. Furthermore, in vitro cytotoxicity assays showed that CUR-loaded ChiLipo–Ser nanoparticles exerted cytotoxic effects against HeLa and HT29 cancer cells while maintaining acceptable biocompatibility in BJ fibroblasts. Collectively, these results highlight the potential of ChiLipo–Ser nanoparticles as a promising platform for stimuli-responsive anticancer drug delivery.
本研究通过α-硫辛酸和l-丝氨酸(ChiLipo-Ser)的双重修饰,构建了一种新型的壳聚糖纳米载体体系,以提高其物理化学和生物学性能,用于药物递送。FTIR和1H NMR验证了壳聚糖骨架上官能团的成功偶联,而GPC分析证实了分子量的增加与改性一致。与天然壳聚糖相比,ChiLipo-Ser在生理pH下的溶解度提高,缓冲能力增强。通过离子化凝胶法制备的纳米颗粒表现出良好的特性,包括平均粒径约150 nm,低多分散性和表面带正电荷。DLS分析证实了氧化还原敏感性,发现在10-20 mM DTT存在下,由于二硫键断裂导致颗粒不稳定。重要的是,与未修饰的壳聚糖相比,ChiLipo-Ser修饰显著提高了姜黄素的包封效率,同时所得到的溶液在pH 5下也表现出有效的释放行为,模拟肿瘤细胞内环境。此外,体外细胞毒性实验表明,cur负载的ChiLipo-Ser纳米颗粒对HeLa和HT29癌细胞具有细胞毒性作用,同时在BJ成纤维细胞中保持可接受的生物相容性。总的来说,这些结果突出了chilibo - ser纳米颗粒作为刺激反应性抗癌药物递送的有前途的平台的潜力。
{"title":"α-lipoic acid and L-serine conjugated chitosan as a stimuli-responsive drug delivery platform","authors":"Özlem Kaplan ,&nbsp;Kevser Bal ,&nbsp;Sibel Küçükertuğrul Çeli̇k ,&nbsp;Nazan Gökşen Tosun ,&nbsp;Mehmet Koray Gök","doi":"10.1016/j.carres.2026.109837","DOIUrl":"10.1016/j.carres.2026.109837","url":null,"abstract":"<div><div>In this study, a novel chitosan-based nanocarrier system was developed through dual modification with α-lipoic acid and <span>l</span>-serine (Chi<sub>Lipo–Ser</sub>) to enhance its physicochemical and biological properties for drug delivery applications. FTIR and <sup>1</sup>H NMR spectroscopy verified the successful conjugation of functional groups onto the chitosan backbone, while GPC analysis confirmed an increase in molecular weight consistent with the modification. Compared with native chitosan, the Chi<sub>Lipo–Ser</sub> exhibited improved solubility at physiological pH and enhanced buffering capacity. Nanoparticles prepared via ionotropic gelation demonstrated favorable characteristics, including an average particle size of ∼150 nm, low polydispersity, and a positive surface charge. Redox sensitivity was evidenced by DLS analysis, which revealed particle destabilization in the presence of 10–20 mM DTT, resulting from disulfide bond cleavage. Importantly, Chi<sub>Lipo–Ser</sub> modification significantly enhanced the encapsulation efficiency of curcumin compared to unmodified chitosan, while the resulting formulations also exhibited effective release behavior at pH 5, mimicking the tumor intracellular environment. Furthermore, <em>in vitro</em> cytotoxicity assays showed that CUR-loaded Chi<sub>Lipo–Ser</sub> nanoparticles exerted cytotoxic effects against HeLa and HT29 cancer cells while maintaining acceptable biocompatibility in BJ fibroblasts. Collectively, these results highlight the potential of Chi<sub>Lipo–Ser</sub> nanoparticles as a promising platform for stimuli-responsive anticancer drug delivery.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"562 ","pages":"Article 109837"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146036015","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Carbohydrate Research
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