Pub Date : 2025-12-12DOI: 10.1016/j.carres.2025.109791
Linhan Wang , Jichao Wei , Fang Yuan , Yongheng Rong , Wanqian Du , Wenjing Shi , Xintan Wang , Mei Wang , Yankang Wang , Anran Liu , Junqiao Zhao , Yun Kong , Na Sun , Wenzhu Tang , Shengjun Wang
β1-3-Galactosyltransferase from Drosophila melanogaster (DmC1GalT1) plays a key role in O-glycan synthesis and holds considerable potential for glycoengineering applications. In this study, we achieved high-yield expression and purification of DmC1GalT1 in Escherichia coli, obtaining over 5 mg of protein per liter of culture. Similar with the enzyme expressed from Pichia pastoris, this enzyme exhibited strict donor specificity towards UDP-Gal and efficiently galactosylated human CD74-derived Tn-glycopeptides. Structural analysis identified key residues involved in substrate binding and catalysis. Site-directed mutagenesis of active-site residues yielded variants with altered activity and stability profiles. Notably, the N108G and Y325W mutants retained high activity towards UDP-Gal but lost the ability to utilize UDP-GalNH2, while N108A mutants retained 61.77 % of its activity toward UDP-Gal but maintained similar activity levels towards UDP-GalNH2, while gaining weak but detectable activity toward UDP-Glc. These findings highlight the critical roles of residues N108 and Y325 in donor recognition. Thermal stability predictions using ProStab indicated that certain mutations affected enzyme stability without compromising activity. This work establishes an efficient platform for the expression, characterization, and engineering of DmC1GalT1, facilitating the development of glycosyltransferase variants with tailored properties for synthetic glycobiology.
{"title":"Characterization and engineering of Drosophila melanogaster β1-3-galactosyltransferase for glycoengineering applications","authors":"Linhan Wang , Jichao Wei , Fang Yuan , Yongheng Rong , Wanqian Du , Wenjing Shi , Xintan Wang , Mei Wang , Yankang Wang , Anran Liu , Junqiao Zhao , Yun Kong , Na Sun , Wenzhu Tang , Shengjun Wang","doi":"10.1016/j.carres.2025.109791","DOIUrl":"10.1016/j.carres.2025.109791","url":null,"abstract":"<div><div>β1-3-Galactosyltransferase from <em>Drosophila melanogaster</em> (<em>Dm</em>C1GalT1) plays a key role in O-glycan synthesis and holds considerable potential for glycoengineering applications. In this study, we achieved high-yield expression and purification of <em>Dm</em>C1GalT1 in <em>Escherichia coli,</em> obtaining over 5 mg of protein per liter of culture. Similar with the enzyme expressed from <em>Pichia pastoris</em>, this enzyme exhibited strict donor specificity towards UDP-Gal and efficiently galactosylated human CD74-derived Tn-glycopeptides. Structural analysis identified key residues involved in substrate binding and catalysis. Site-directed mutagenesis of active-site residues yielded variants with altered activity and stability profiles. Notably, the N108G and Y325W mutants retained high activity towards UDP-Gal but lost the ability to utilize UDP-GalNH<sub>2</sub>, while N108A mutants retained 61.77 % of its activity toward UDP-Gal but maintained similar activity levels towards UDP-GalNH<sub>2</sub>, while gaining weak but detectable activity toward UDP-Glc. These findings highlight the critical roles of residues N108 and Y325 in donor recognition. Thermal stability predictions using ProStab indicated that certain mutations affected enzyme stability without compromising activity. This work establishes an efficient platform for the expression, characterization, and engineering of <em>Dm</em>C1GalT1, facilitating the development of glycosyltransferase variants with tailored properties for synthetic glycobiology.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109791"},"PeriodicalIF":2.5,"publicationDate":"2025-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145755350","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}
The glycoside hydrolase family 30 (GH30) enzyme RfGH30, originating from the ruminal bacterium Ruminococcus flavefaciens, was biochemically characterized to elucidate its catalytic properties and substrate specificity. Although initially classified as endoxylanase, experimental analysis confirmed RfGH30 as processive exo-β-1,6-galactanase targeting galactan-rich polysaccharides. Zymogram analysis revealed a clear hydrolytic zone at ∼54 kDa on larchwood arabinogalactan (LWAG), confirming its specificity for β-1,6-glycosidic linkage containing galactose units. The enzyme exhibited optimum pH 5.5 and temperature 40 °C, retaining stability across pH 3.8–7.0 and temperature range, 4–70 °C. Among natural substrates, RfGH30 displayed highest specific activity, 8.1 U mg−1 against LWAG followed by arabinan 4.6 U mg−1 against sugarbeet arabinan. RfGH30 also hydrolysed synthetic substrates, pNPG and oNPG consistent with an exo-mode of catalysis. Kinetic analysis showed a Vmax of 9.8 U mg−1 and KM of 1.45 mg mL−1 against LWAG. Enzyme activity was enhanced by Ca2+ and Mg2+, but inhibited by Fe2+ and the chelating agent EGTA. RfGH30 hydrolysed LWAG product profile by TLC, HPLC and HRMS revealed the predominant release of β-1,6-galactobiose along with minor galactose, demonstrating strict cleavage of β-1,6-galactosyl linkages. RfGH30 showed strong processivity with LWAG, with the high processivity index of 21 and a low, 4.7 with sugarbeet arabinan. This study reports the first biochemical evidence of a GH30 processive exo-β-1,6-galactanase from R. flavefaciens, providing insights into its role in ruminal degradation of plant arabinogalactans and its potential applications in biomass valorization.
糖苷水解酶家族30 （GH30）酶RfGH30源自瘤胃细菌黄瘤球菌，对其进行了生化表征，以阐明其催化特性和底物特异性。虽然最初被归类为内木聚糖酶，但实验分析证实RfGH30是针对富含半乳糖多糖的显性外显子-β-1,6-半乳糖酶。酶谱分析显示，落叶松阿拉伯半乳糖（LWAG）在~ 54 kDa处有一个清晰的水解区，证实了它对含有半乳糖单元的β-1,6-糖苷连锁的特异性。该酶在pH为5.5、温度为40℃时表现出最佳稳定性，在pH为3.8 ~ 7.0、温度为4 ~ 70℃范围内均保持稳定。在天然底物中，RfGH30对LWAG的比活性最高，为8.1 U mg-1，其次是arabinan，对甜菜arabinan的比活性为4.6 U mg-1。RfGH30还水解合成底物pNPG和oNPG，符合外显催化模式。动力学分析表明，对LWAG的Vmax为9.8 U mg-1， KM为1.45 mg mL-1。Ca2+和Mg2+对酶活性有增强作用，Fe2+和EGTA对酶活性有抑制作用。通过TLC、HPLC和HRMS分析，RfGH30水解后的LWAG产物主要释放β-1,6-半乳糖糖和少量半乳糖，显示了β-1,6-半乳糖键的严格断裂。RfGH30对LWAG的加工性较强，加工性指数高，为21，对阿拉伯甜菜聚糖的加工性指数低，为4.7。本研究首次报道了黄芽霉GH30进程性外显-β-1,6-半乳糖酶的生化证据，为其在植物阿拉伯半乳糖瘤胃降解中的作用及其在生物质增值中的潜在应用提供了新的见解。
{"title":"Uncovering the β-1,6-galactan-degrading machinery: Biochemical and functional insights into a processive exo-β-1,6-galactanase, RfGH30 from Ruminococcus flavefaciens","authors":"Shreya Biswas, Gurleen Kaur Bagga, Parmeshwar Vitthal Gavande , Arun Goyal","doi":"10.1016/j.carres.2025.109793","DOIUrl":"10.1016/j.carres.2025.109793","url":null,"abstract":"<div><div>The glycoside hydrolase family 30 (GH30) enzyme <em>Rf</em>GH30, originating from the ruminal bacterium <em>Ruminococcus flavefaciens</em>, was biochemically characterized to elucidate its catalytic properties and substrate specificity. Although initially classified as endoxylanase, experimental analysis confirmed <em>Rf</em>GH30 as processive exo-β-1,6-galactanase targeting galactan-rich polysaccharides. Zymogram analysis revealed a clear hydrolytic zone at ∼54 kDa on larchwood arabinogalactan (LWAG), confirming its specificity for β-1,6-glycosidic linkage containing galactose units. The enzyme exhibited optimum pH 5.5 and temperature 40 °C, retaining stability across pH 3.8–7.0 and temperature range, 4–70 °C. Among natural substrates, <em>Rf</em>GH30 displayed highest specific activity, 8.1 U mg<sup>−1</sup> against LWAG followed by arabinan 4.6 U mg<sup>−1</sup> against sugarbeet arabinan. <em>Rf</em>GH30 also hydrolysed synthetic substrates, <em>p</em>NPG and <em>o</em>NPG consistent with an exo-mode of catalysis. Kinetic analysis showed a <em>V</em><sub>max</sub> of 9.8 U mg<sup>−1</sup> and <em>K</em><sub><em>M</em></sub> of 1.45 mg mL<sup>−1</sup> against LWAG. Enzyme activity was enhanced by Ca<sup>2+</sup> and Mg<sup>2+</sup>, but inhibited by Fe<sup>2+</sup> and the chelating agent EGTA. <em>Rf</em>GH30 hydrolysed LWAG product profile by TLC, HPLC and HRMS revealed the predominant release of β-1,6-galactobiose along with minor galactose, demonstrating strict cleavage of β-1,6-galactosyl linkages. <em>Rf</em>GH30 showed strong processivity with LWAG, with the high processivity index of 21 and a low, 4.7 with sugarbeet arabinan. This study reports the first biochemical evidence of a GH30 processive exo-β-1,6-galactanase from <em>R. flavefaciens</em>, providing insights into its role in ruminal degradation of plant arabinogalactans and its potential applications in biomass valorization.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109793"},"PeriodicalIF":2.5,"publicationDate":"2025-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145755373","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}
Carboxymethyl starch (CMSt) is a modified starch with excellent performance. In this study, sodium carbonate pretreatment was used to promote the carboxymethylation reaction activity of starch, carboxymethyl starch with medium and low substitution degrees was successfully prepared, and the synthesis parameters of carboxymethyl starch were optimized using response surface methodology. The results showed that when the pretreatment concentration was 0.95 mol/L, the pretreatment time was 54 min, the molar ratio of alkali to starch was 1.3, and the molar ratio of acid to starch was 1, the carboxymethyl starch with a degree of substitution (DS) of 0.284 was obtained, which was 123.62 % higher than that without of the pretreatment process. The prepared carboxymethyl starch was used to encapsulate acai berry anthocyanins (ABA) in microcapsules, and the biomedical application of the prepared carboxymethyl starch was initially attempted. This study provided a new method for preparing carboxymethyl starch and attempted to explore the application value of this product in biomedicine.
{"title":"Preparation of carboxymethyl potato starch using a new sodium carbonate pretreatment process and its microencapsulation application","authors":"Yong Chen, Cheng Tang, Ziqiang Zhao, Zhimin Liu, Jiajia Huang, Qijie Chen","doi":"10.1016/j.carres.2025.109790","DOIUrl":"10.1016/j.carres.2025.109790","url":null,"abstract":"<div><div>Carboxymethyl starch (CMSt) is a modified starch with excellent performance. In this study, sodium carbonate pretreatment was used to promote the carboxymethylation reaction activity of starch, carboxymethyl starch with medium and low substitution degrees was successfully prepared, and the synthesis parameters of carboxymethyl starch were optimized using response surface methodology. The results showed that when the pretreatment concentration was 0.95 mol/L, the pretreatment time was 54 min, the molar ratio of alkali to starch was 1.3, and the molar ratio of acid to starch was 1, the carboxymethyl starch with a degree of substitution (DS) of 0.284 was obtained, which was 123.62 % higher than that without of the pretreatment process. The prepared carboxymethyl starch was used to encapsulate acai berry anthocyanins (ABA) in microcapsules, and the biomedical application of the prepared carboxymethyl starch was initially attempted. This study provided a new method for preparing carboxymethyl starch and attempted to explore the application value of this product in biomedicine.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109790"},"PeriodicalIF":2.5,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733699","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}
Pub Date : 2025-12-09DOI: 10.1016/j.carres.2025.109788
Imlirenla Pongener , Gavin J. Miller
{"title":"Exploration of a heparosan trisaccharide synthesis: donor evaluation and characterisation of an unexpected glucuronic acid acceptor pyranose to furanose ring contraction","authors":"Imlirenla Pongener , Gavin J. Miller","doi":"10.1016/j.carres.2025.109788","DOIUrl":"10.1016/j.carres.2025.109788","url":null,"abstract":"","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109788"},"PeriodicalIF":2.5,"publicationDate":"2025-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145773662","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}
Pub Date : 2025-12-04DOI: 10.1016/j.carres.2025.109780
Wanjun Jin , Cheng Li , Chengjian Wang , Ming Wei , Yuanlin Sun , Qingzhen Yang , Zhongfu Wang , Linjuan Huang
N/O-glycans in various human seminal plasma (hSP) glycoproteins, especially α2,3- and α2,6-sialylation levels, are closely associated with semen quality. However, effective differentiation of sialyl linkage isomer remains unachieved. Here, we employed our previously developed glycoqueuing strategy for isomer-specific quantitative analysis of sialylated N/O-glycans released from hSP. A total of 21 exclusively α2,6-sialylated and 14 α2,3-sialylated (bearing α2,3-linked sialic acids) N-glycan isomers were detected, and the relative abundance of α2,6-sialylation among these sialylated N-glycans reached 61.40 %. For O-glycans, seven monosialic and five disialylated species were observed, and all were confirmed to be α2,3-sialylated. Nonsialylated N/O-glycan isomers were simultaneously quantified via hydrophilic interaction liquid chromatography-tandem mass spectrometry. Specifically, 21 nonsialylated N-glycan isomers were identified, among which the relative abundances of oligomannose-type and complex-type glycan were each approximately 50 %. All O-glycans exhibited core 1 or core 2 structures, including 12 α2,3-sialylated and 26 nonsialylated species. Notably, four isomers separated from two newly discovered nonsialylated O-glycan compositions H2N1F1 and H2N1F2 (H: hexose, N: N-acetylgalactosamine, F: fucose) were identified in hSP. Additionally, sialylated and nonsialylated N/O-glycans were all highly fucosylated (16.98 %–67.92 %), and bore numerous Lewis X and Lewis Y structures. The detailed glycan structural and distribution data, particularly α2,3/α2,6-sialylation profiles, not only provides a reference for constructing the hSP glycomic fingerprint, but also supports in-depth investigation of hSP glycan functions in reproduction and exploration of novel glycan markers for clinical semen quality detection.
{"title":"Differentiation of α2,3- and α2,6-linked sialylated N/O-glycan isomers in human seminal plasma by an glycoqueuing strategy","authors":"Wanjun Jin , Cheng Li , Chengjian Wang , Ming Wei , Yuanlin Sun , Qingzhen Yang , Zhongfu Wang , Linjuan Huang","doi":"10.1016/j.carres.2025.109780","DOIUrl":"10.1016/j.carres.2025.109780","url":null,"abstract":"<div><div><em>N/O</em>-glycans in various human seminal plasma (hSP) glycoproteins, especially α2,3- and α2,6-sialylation levels, are closely associated with semen quality. However, effective differentiation of sialyl linkage isomer remains unachieved. Here, we employed our previously developed glycoqueuing strategy for isomer-specific quantitative analysis of sialylated <em>N/O-</em>glycans released from hSP. A total of 21 exclusively α2,6-sialylated and 14 α2,3-sialylated (bearing α2,3-linked sialic acids) <em>N-</em>glycan isomers were detected, and the relative abundance of α2,6-sialylation among these sialylated <em>N</em>-glycans reached 61.40 %. For <em>O</em>-glycans, seven monosialic and five disialylated species were observed, and all were confirmed to be α2,3-sialylated. Nonsialylated <em>N/O-</em>glycan isomers were simultaneously quantified via hydrophilic interaction liquid chromatography-tandem mass spectrometry. Specifically, 21 nonsialylated <em>N-</em>glycan isomers were identified, among which the relative abundances of oligomannose-type and complex-type glycan were each approximately 50 %. All <em>O-</em>glycans exhibited core 1 or core 2 structures, including 12 α2,3-sialylated and 26 nonsialylated species. Notably, four isomers separated from two newly discovered nonsialylated <em>O</em>-glycan compositions H2N1F1 and H2N1F2 (H: hexose, N: <em>N-</em>acetylgalactosamine, F: fucose) were identified in hSP. Additionally, sialylated and nonsialylated <em>N/O-</em>glycans were all highly fucosylated (16.98 %–67.92 %), and bore numerous Lewis X and Lewis Y structures. The detailed glycan structural and distribution data, particularly α2,3/α2,6-sialylation profiles, not only provides a reference for constructing the hSP glycomic fingerprint, but also supports in-depth investigation of hSP glycan functions in reproduction and exploration of novel glycan markers for clinical semen quality detection.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109780"},"PeriodicalIF":2.5,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145682905","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}
We report herein the olefination of a set of six N-Boc glyconolactams using Petasis reagent to yield unprecedented exoiminoglycals. In addition, a modified Simmons-Smith cyclopropanation was successfully applied to the D-gluco-configured olefin to furnish the unprecedented 1-spirocyclic deoxynojirimycin which proved to be a poor inhibitor of glycosidases in its unprotected form.
{"title":"Petasis-mediated exo-iminoglycals synthesis from glyconolactams allows access to unprecedented 1-spirocyclopropyl deoxynojirimycin","authors":"Lilou Chopin , Aurélien Beato , Dylan Yorga , Nicolas Auberger , Jean-Bernard Behr , Jérôme Désiré , Yves Blériot","doi":"10.1016/j.carres.2025.109765","DOIUrl":"10.1016/j.carres.2025.109765","url":null,"abstract":"<div><div>We report herein the olefination of a set of six <em>N</em>-Boc glyconolactams using Petasis reagent to yield unprecedented exoiminoglycals. In addition, a modified Simmons-Smith cyclopropanation was successfully applied to the D-<em>gluco</em>-configured olefin to furnish the unprecedented 1-spirocyclic deoxynojirimycin which proved to be a poor inhibitor of glycosidases in its unprotected form.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109765"},"PeriodicalIF":2.5,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733698","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}
Pub Date : 2025-12-03DOI: 10.1016/j.carres.2025.109787
Tiangui Wu , Pengfei Ye , Junjie Yang , Yuhan Sun
N-glycosylation is a dynamic post-translational modification that critically regulates cancer cell differentiation through modulating receptor signaling, cell adhesion, and plasticity. Aberrant N-glycosylation promotes dedifferentiation, drives EMT, and confers therapy resistance across malignancies. This review summarizes the role of N-glycosylation in determining lineage commitment and altering responses to differentiation therapies. Targeting the N-glycosylation apparatus can reprogram tumor cells toward differentiated phenotypes, potentiating the effects of agents such as ATRA and NaBu. Evidence from leukemia and solid tumors reveals the therapeutic potential of disrupting glycan-dependent cell fate decisions. Deciphering these “glycan codes” provides a framework for integrating glycosylation modifiers into precision differentiation therapies, offering novel strategies to overcome treatment resistance.
n -糖基化是一种动态的翻译后修饰，通过调节受体信号、细胞粘附和可塑性来关键地调节癌细胞分化。异常的n -糖基化促进去分化，驱动EMT，并赋予恶性肿瘤治疗抵抗。本文综述了n -糖基化在决定谱系承诺和改变分化治疗反应中的作用。靶向n -糖基化装置可以将肿瘤细胞重编程为分化表型，从而增强ATRA和NaBu等药物的作用。来自白血病和实体瘤的证据揭示了破坏聚糖依赖细胞命运决定的治疗潜力。破译这些“聚糖编码”为将糖基化修饰剂整合到精确分化治疗中提供了一个框架，为克服治疗耐药性提供了新的策略。
{"title":"Emerging insights into cell differentiation: the role of N-glycosylation in differentiation-based cancer therapies","authors":"Tiangui Wu , Pengfei Ye , Junjie Yang , Yuhan Sun","doi":"10.1016/j.carres.2025.109787","DOIUrl":"10.1016/j.carres.2025.109787","url":null,"abstract":"<div><div><em>N</em>-glycosylation is a dynamic post-translational modification that critically regulates cancer cell differentiation through modulating receptor signaling, cell adhesion, and plasticity. Aberrant <em>N</em>-glycosylation promotes dedifferentiation, drives EMT, and confers therapy resistance across malignancies. This review summarizes the role of <em>N</em>-glycosylation in determining lineage commitment and altering responses to differentiation therapies. Targeting the <em>N</em>-glycosylation apparatus can reprogram tumor cells toward differentiated phenotypes, potentiating the effects of agents such as ATRA and NaBu. Evidence from leukemia and solid tumors reveals the therapeutic potential of disrupting glycan-dependent cell fate decisions. Deciphering these “glycan codes” provides a framework for integrating glycosylation modifiers into precision differentiation therapies, offering novel strategies to overcome treatment resistance.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109787"},"PeriodicalIF":2.5,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145682906","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}
Pub Date : 2025-12-01DOI: 10.1016/j.carres.2025.109785
Matej Cvečko, Vladimír Mastihuba, Mária Mastihubová
Enzymatic methods leading to the preparation of biologically active glycophenols are attracting increasing interest due to their selectivity and sustainability. In this study, a series of hydrophilic and hydrophobic 4-O-acetylferulic acid sugar esters was prepared using a combination of chemical and enzymatic approaches. During the investigation of chemoselective deacetylation of the phenolic groups in these esters, a pronounced cooperative effect between Lipase PS and bovine serum albumin (BSA) was observed in a biphasic MTBE–water system. In the presence of BSA, Lipase PS more rapidly and selectively deacetylated the phenolic acetyl group of hydrophobic substrates. In contrast, no rate enhancement was observed for more polar substrates bearing free hydroxyl groups. Protecting groups on the carbohydrate moiety remained unaffected, and the ester bond between ferulic acid and the sugar was preserved. The accelerating effect of BSA on hydrophobic substrates is attributed to its surface-active properties, which increase the interfacial area through the formation of stable emulsions. Given that the intrinsic acetylesterase activity of BSA is approximately 300-fold lower than that of Lipase PS, BSA alone is not capable of effective deacetylation. These findings highlight the potential of surface-active proteins to improve biocatalytic transformations of hydrophobic substrates while avoiding purification challenges associated with low-molecular-weight surfactants.
酶法制备具有生物活性的糖酚由于其选择性和可持续性而引起越来越多的兴趣。本研究采用化学和酶相结合的方法制备了一系列亲水和疏水的4- o -乙酰阿魏酸糖酯。在两相mtbe -水体系中，脂肪酶PS与牛血清白蛋白（BSA）有明显的协同作用。在BSA存在的情况下，脂肪酶PS能更快、更有选择性地使疏水底物的酚乙酰基脱乙酰化。相比之下，没有观察到更多极性底物携带游离羟基的速率增强。碳水化合物部分的保护基团不受影响，阿魏酸和糖之间的酯键被保留了下来。BSA在疏水基质上的加速作用是由于其表面活性，通过形成稳定的乳剂来增加界面面积。鉴于牛血清白蛋白的内在乙酰酯酶活性比脂肪酶PS低约300倍，单靠牛血清白蛋白不能有效地去乙酰化。这些发现强调了表面活性蛋白在改善疏水底物生物催化转化方面的潜力，同时避免了与低分子量表面活性剂相关的纯化挑战。
{"title":"Chemoselective deacetylation of hydrophobic glycophenols by lipase PS enhanced with bovine serum albumin","authors":"Matej Cvečko, Vladimír Mastihuba, Mária Mastihubová","doi":"10.1016/j.carres.2025.109785","DOIUrl":"10.1016/j.carres.2025.109785","url":null,"abstract":"<div><div>Enzymatic methods leading to the preparation of biologically active glycophenols are attracting increasing interest due to their selectivity and sustainability. In this study, a series of hydrophilic and hydrophobic 4-<em>O</em>-acetylferulic acid sugar esters was prepared using a combination of chemical and enzymatic approaches. During the investigation of chemoselective deacetylation of the phenolic groups in these esters, a pronounced cooperative effect between Lipase PS and bovine serum albumin (BSA) was observed in a biphasic MTBE–water system. In the presence of BSA, Lipase PS more rapidly and selectively deacetylated the phenolic acetyl group of hydrophobic substrates. In contrast, no rate enhancement was observed for more polar substrates bearing free hydroxyl groups. Protecting groups on the carbohydrate moiety remained unaffected, and the ester bond between ferulic acid and the sugar was preserved. The accelerating effect of BSA on hydrophobic substrates is attributed to its surface-active properties, which increase the interfacial area through the formation of stable emulsions. Given that the intrinsic acetylesterase activity of BSA is approximately 300-fold lower than that of Lipase PS, BSA alone is not capable of effective deacetylation. These findings highlight the potential of surface-active proteins to improve biocatalytic transformations of hydrophobic substrates while avoiding purification challenges associated with low-molecular-weight surfactants.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109785"},"PeriodicalIF":2.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145667203","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}
Pub Date : 2025-12-01DOI: 10.1016/j.carres.2025.109786
Mingyang Cao , Ruisen Sun , Hong Dongchu , Jiaxin Lin , Yue Zhang , Zhao Feiya , Tao Aien
Cornus officinalis (C. officinalis) is an important plant resource with a long history and a wide range of uses in medicine and food. Polysaccharides are one of the main active components of C. officinalis. Because of its unique and significant anti-tumor, antioxidant, immunomodulatory, and many other biological activities, it has received extensive attention from researchers, and its study has become more and more in-depth. Although the phytochemical and bioactive aspects of C. officinalis polysaccharides (COPs) have been extensively studied, a systematic and comprehensive summary of these polysaccharides has not yet been compiled. This lack of summary hinders their full utilization and development. This paper reviews the extraction, purification, structural features, and biological activities, as well as applications of COPs. It also discusses the potential development and future research directions of COPs in the food, pharmaceutical, and cosmeceutical fields. This paper may provide direction and a theoretical basis for the development of COPs as novel functional foods.
{"title":"Extraction, purification, structural characterization, biological activities, applications, and research prospects of Cornus officinalis polysaccharides: a review","authors":"Mingyang Cao , Ruisen Sun , Hong Dongchu , Jiaxin Lin , Yue Zhang , Zhao Feiya , Tao Aien","doi":"10.1016/j.carres.2025.109786","DOIUrl":"10.1016/j.carres.2025.109786","url":null,"abstract":"<div><div><em>Cornus officinalis</em> (<em>C. officinalis</em>) is an important plant resource with a long history and a wide range of uses in medicine and food. Polysaccharides are one of the main active components of <em>C. officinalis</em>. Because of its unique and significant anti-tumor, antioxidant, immunomodulatory, and many other biological activities, it has received extensive attention from researchers, and its study has become more and more in-depth. Although the phytochemical and bioactive aspects of <em>C. officinalis</em> polysaccharides (COPs) have been extensively studied, a systematic and comprehensive summary of these polysaccharides has not yet been compiled. This lack of summary hinders their full utilization and development. This paper reviews the extraction, purification, structural features, and biological activities, as well as applications of COPs. It also discusses the potential development and future research directions of COPs in the food, pharmaceutical, and cosmeceutical fields. This paper may provide direction and a theoretical basis for the development of COPs as novel functional foods.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109786"},"PeriodicalIF":2.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145720969","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}
Pub Date : 2025-11-29DOI: 10.1016/j.carres.2025.109781
Yue Zhu , Can Jin , Kan Ding
As targeted therapy assumes an increasingly pivotal role in comprehensive cancer treatment, there is a growing imperative to develop natural drugs that exhibit low toxicity and minimal side effects. Polysaccharides derived from the traditional Chinese herbal medicine Arisaema erubescens have been reported to show antitumor potential, however the key structural features and specific molecular targets responsible for their pharmacological effects are still vague. To address this question, a homogeneous polysaccharide TNX05 (Mw ≈ 9 kDa) was obtained and characterized from Arisaema erubescens. Structural analysis suggested that TNX05 was a galactoglucan with a backbone of 1, 4-α-D-Glcp, 1, 4-β-D-Glcp, and 1, 3-β-D-Galp residues, with side chains—→1-α/β-D-Glcp-(6 → 1)-α-D-Glcp and →1)-α-D-Glcp—substituted at the C-4 and C-6 of the 1, 4-α-D-Glcp units, respectively. Combining enzymatic hydrolysis, molecular docking, and protein-binding assays we showed a key core domain (TNX05II), which exhibited micromolar-range binding affinity for ten tumor-associated targets: Glypican-6 (GPC-6), glucuronic acid epimerase (Glce), S100 calcium-binding protein A4 (S100A4), S100A6, nucleoside diphosphate kinase 1 (NME1), fibroblast growth factor 17 (FGF17), proto-oncogene tyrosine-protein kinase Src (SRC), kelch-like ECH-associated protein 1 (KEAP1), Galectin-3 (Gal-3), and protein phosphatase 3 catalytic subunit alpha (PPP3CA). Additionally, TNX05II was significantly resistant to α-glucosidase degradation. This study suggests a possible key structure-target relationship underlying TNX05's antitumor activity, providing a molecular basis for the antitumor mechanism of Arisaema polysaccharides.
随着靶向治疗在癌症综合治疗中发挥越来越重要的作用，开发低毒、副作用小的天然药物势在必行。从传统中草药鸢尾中提取的多糖已被报道显示出抗肿瘤的潜力，但其药理作用的关键结构特征和特定的分子靶点仍不清楚。为了解决这一问题，从鸢尾中分离得到了一种均质多糖TNX05 (Mw≈9 kDa)，并对其进行了表征。结构分析表明，TNX05是一种以1,4 -α- d - glcp、1,4 -β-D-Glcp和1,3 -β- d - galp为骨架的半乳糖葡聚糖，侧链-→1-α/β-D-Glcp-(6→1)-α- d - glcp和→1)-α- d - glcp -分别取代1,4 -α- d - glcp单元的C-4和C-6。结合酶解、分子对接和蛋白质结合实验，我们发现了一个关键的核心结构域（TNX05II），它对10个肿瘤相关靶点具有微摩尔范围的结合亲和力：Glypican-6 （GPC-6）、葡萄糖醛酸epimase （Glce）、S100钙结合蛋白A4 （S100A4）、S100A6、核苷二磷酸激酶1 （NME1）、成纤维细胞生长因子17 （FGF17）、原癌基因酪氨酸蛋白激酶Src （Src）、kelch样ech相关蛋白1 （KEAP1）、半乳糖凝集素-3 （Gal-3）和蛋白磷酸酶3催化亚基α (PPP3CA)。此外，TNX05II对α-葡萄糖苷酶降解具有显著抗性。本研究提示了TNX05抗肿瘤活性可能存在的关键结构-靶点关系，为菝葜多糖抗肿瘤机制的研究提供了分子基础。
{"title":"A structurally defined galactoglucan from Arisaema erubescens with a multi-target tumor-associated protein binding domain","authors":"Yue Zhu , Can Jin , Kan Ding","doi":"10.1016/j.carres.2025.109781","DOIUrl":"10.1016/j.carres.2025.109781","url":null,"abstract":"<div><div>As targeted therapy assumes an increasingly pivotal role in comprehensive cancer treatment, there is a growing imperative to develop natural drugs that exhibit low toxicity and minimal side effects. Polysaccharides derived from the traditional Chinese herbal medicine <em>Arisaema erubescens</em> have been reported to show antitumor potential, however the key structural features and specific molecular targets responsible for their pharmacological effects are still vague. To address this question, a homogeneous polysaccharide TNX05 (<em>Mw</em> ≈ 9 kDa) was obtained and characterized from <em>Arisaema erubescens</em>. Structural analysis suggested that TNX05 was a galactoglucan with a backbone of 1, 4-α-<sub>D</sub>-Glc<em>p</em>, 1, 4-β-<sub>D</sub>-Glc<em>p</em>, and 1, 3-β-<sub>D</sub>-Gal<em>p</em> residues, with side chains—→1-α/β-<sub>D</sub>-Glc<em>p</em>-(6 → 1)-α-<sub>D</sub>-Glc<em>p</em> and →1)-α-<sub>D</sub>-Glc<em>p</em>—substituted at the C-4 and C-6 of the 1, 4-α-<sub>D</sub>-Glc<em>p</em> units, respectively. Combining enzymatic hydrolysis, molecular docking, and protein-binding assays we showed a key core domain (TNX05II), which exhibited micromolar-range binding affinity for ten tumor-associated targets: Glypican-6 (GPC-6), glucuronic acid epimerase (Glce), S100 calcium-binding protein A4 (S100A4), S100A6, nucleoside diphosphate kinase 1 (NME1), fibroblast growth factor 17 (FGF17), proto-oncogene tyrosine-protein kinase Src (SRC), kelch-like ECH-associated protein 1 (KEAP1), Galectin-3 (Gal-3), and protein phosphatase 3 catalytic subunit alpha (PPP3CA). Additionally, TNX05II was significantly resistant to α-glucosidase degradation. This study suggests a possible key structure-target relationship underlying TNX05's antitumor activity, providing a molecular basis for the antitumor mechanism of Arisaema polysaccharides.</div></div>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"560 ","pages":"Article 109781"},"PeriodicalIF":2.5,"publicationDate":"2025-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145676638","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}
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