GLYCOBIOLOGY
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2016
·Vol. 26
·期 8
·871-879
·
IF 3.283
Liu Si,Kulinich Anna,Cai Zhi P,Ma Hong Y,Du Ya M,Lv Yong M,Liu Li,Voglmeir Josef
Three novel bacterial α-l-fucosidases, which cleave terminal fucosyl residues from glycoconjugates are reported in this work. Originating from the recently discovered bacterium Emticicia oligotrophica, recombinant fucosidase isoforms designated as Eo0918, Eo3066 and Eo3812 were shown to have the highest activity between pH 6.0 and 7.0 and temperature optima between 30 and 45°C. All enzymes catalyzed the hydrolysis of the model substrate pNP-α-l-fucose and revealed significantly different regiospecificities towards fucose-containing oligosaccharides: Eo0918 liberated exclusively α1,6-linked fucose and Eo3812 released only α1,3-fucosyl residues, whereas Eo3066 showed broader substrate promiscuity. The enzymatic activity of Eo0918 and Eo3812 increased upon the addition of Ca2+, Mn2+ and Zn2+ ions, whereas the activity of Eo3066 was significantly decreased in the presence of these metal ions. In addition, Eo0918 also catalyzed the transfer of fucose from pNP-α-l-fucose to the 7-hydroxyl group of 4-methylumbelliferone with up to 15% transglycosylation yield. Facile recombinant expression in E. coli, distinct substrate specificities and the transglycosylation ability of Eo0918 presented herein make these newly discovered fucosidases valuable candidates for bioanalytical and biotechnological applications.
研究领域:
微生物学
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生物化学
·
生物技术
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DOI
PubMed
高影响因子
PLoS Genetics
·
2025
·Vol. 21
·期 8
·e1011815
·
IF 3.7
Qinqin Zhao,Liwei Gao,Nuo Xu,Xiuting Zhang,Yuqi Qin,Yinbo Qu,Guodong Liu
l -Fucose is a universal capping component of biomolecules found throughout all domains of life. Although fungi are renowned for their role in biomass recycling, the mechanisms by which they process l -fucose remain largely unknown. In this study, we elucidate a l -fucose-responsive system in Trichoderma reesei , a model fungus for plant cell wall degradation. Central to this system is the transcription factor FUR1, which is indispensable for growth on l -fucose. FUR1 orchestrates the expression of l -fucose catabolic enzymes, including an l -fucose dehydrogenase that exhibits distant homology to counterparts in bacteria and mammals. Through RNA sequencing and biochemical assays, we demonstrate that FUR1 also governs the enzymatic liberation of l -fucose by upregulating extracellular α- l -fucosidases. Intriguingly, FUR1 mediates l -fucose-triggered expression of a broad spectrum of enzymes that target diverse glycosidic bonds (e.g., β-glucuronidic, α-galactosidic, and β-xylosidic linkages) within complex carbohydrates. Expression of a constitutively active FUR1 mutant unlocked the production of otherwise silent glycosidases, substantially boosting the hydrolytic capacity of the fungal secretome on orange peel. These findings offer the first molecular insight into l -fucose sensing and metabolism in fungi, and advance our understanding of the fungal regulatory network for coordinated expression of biomass-degrading enzymes.
l -Fucose, is a widely present constituent of polysaccharides, glycoproteins, and glycolipids in plants, animals and bacteria. l -Fucose frequently presents itself as a terminal monosaccharide, and plays critical roles in cell communication, immunity and host-microbe interactions. While fungi are well-known for breaking down various biomass molecules (e.g., lignocellulose) in nature, their metabolic and regulatory machineries for l -fucose utilization are enigmatic. Here, we examine the transcriptional response of Trichoderma reesei to l -fucose. We discovered that an orphan transcription factor activates the expression of l -fucose catabolic enzymes as well as extracellular α- l -fucosidases. Additionally, we found that this transcription factor also mediates the cross-induction by l -fucose of a wide range of enzymes that break down different types of glycosidic bonds. These findings provide the first understanding of how a fungus senses and metabolize l -fucose, and shed light on the complex regulatory systems that allow fungi to efficiently degrade biomass. This knowledge could help improve industrial fungal strains for the production of more efficient biomass-degrading enzymes.
研究领域:
真菌学
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分子生物学
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生物化学
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生物技术
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基因组学
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微生物生理学
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产品关联
DOI
PubMed
高影响因子
BIOMACROMOLECULES
·
2019
·
IF 5.667
Fei Fan,Ping Zhang,Lihao Wang,Tiantian Sun,Chao Cai,Guangli Yu
Fucosylated chondroitin sulfate (fCS), a representative marine polysaccharide isolated from sea cucumber, possesses diverse biological functions especially as a promising anticoagulant. However, its supply suffers from the challenges of high-cost materials, different species, and batch-to-batch variability. In the present study, we designed a concise route for the synthesis of functional glycomimetics by natural fCS as a template. 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride-mediated amidation was applied on chondroitin sulfates for site-selective alkynylation with controllable ratios between 0.15 and 0.78. A small library of 12 fCS glycomimetics with specific sulfation patterns and fucose branches was prepared through copper-catalyzed azide–alkyne cycloaddition, which was fully characterized by nuclear magnetic resonance spectroscopy and size-exclusion chromatography with multiangle light scattering and refractive index. Through screening of their biological activities, CSE-F1 and CSE-SF1 exhibited anticoagulant activities through intrinsic pathway and inhibition of factor Xa by antithrombin III. The concise approach developed herein supplies novel glycopolymers to mimic the distinct functions of natural polysaccharides and promote the development of marine carbohydrate-based drugs.
研究领域:
海洋化学
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药学
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碳水化合物化学
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产品关联
DOI
PubMed
高影响因子
Systems Microbiology and Biomanufacturing
·
2025
·
IF 2.9
Li Zihan,Zhao Guihong,Zhang Dezhi,Tang Yaqun,Liu Geer,Wang Xiaoyuan
2′-Fucosyllactose is the most abundant human milk oligosaccharides and one of the three essential nutrients for infant growth. Corynebacterium glutamicum is one of the most common industrial fermentation bacteria but cannot synthesize 2′-fucosyllactose. In this study, C. glutamicum ATCC13032 was engineered for 2′-fucosyllactose production from fucose and lactose. The gene futC from Helicobacter pylori encoding α-1,2-fucosyltransferase was codon optimized and mutated at four amino acids (F40S/Q150H/C151R/Q239S). The modified gene futC and the gene fkp from Bacteroides thetaiotaomicron encoding fucokinase/GDP-fucose pyrophosphorylase were overexpressed in plasmid pEC and transformed into C. glutamicum , resulting in CW002. CW002 did not synthesize 2′-fucosyllactose possibly because the substrates fucose and/or lactose did not pass through the cell membrane. Therefore, the gene lacY encoding lactose permease and the gene fucP encoding fucose permease from Escherichia coli were overexpressed in plasmid pXTuf and transformed into CW002, resulting in CW006. CW006 did synthesize 2′-fucosyllactose as expected. It is interesting that the production of 2′-fucosyllactose was decreased or stopped when the expression combination of these four genes was changed, suggesting that the expression levels of the four genes in CW006 might have to well balanced. C. glutamicum CW006 produced 2.07 g/L 2′-fucosyllactose in a 2.4 L bioreactor.
研究领域:
微生物学
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生物工程
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分子生物学
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食品科学
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2019
·
IF 3.571
Yu Lu,Jie Liu,Yue Jia,Yuerong Yang,Qinghui Chen,Lujia Sun,Shuang Song,Linjuan Huang,Zhongfu Wang
Human milk oligosaccharides are complex carbohydrates with multibiofunctional health benefits to newborns. Human milk free oligosaccharides (HMOs) are well characterized. However, changes in the N/O-glycome during lactation are poorly reported. Herein, we qualitatively and quantitatively investigated N/O-glycome profiles and their alteration in human milk at different lactation stages. N-Glycans were mainly fucosylated and nonsialylated, nonfucosylated throughout lactation. O-Glycans mainly consisted of sialylated and nonsialylated, nonfucosylated in colostrum and transitional milk, and fucosylated and nonfucosylated, nonsialylated in mature milk. Fucosylated and sialylated N-glycans gradually decreased and increased, respectively, as lactation progressed; O-glycans showed the reverse. Interestingly, changes in HMO abundance decreased during lactation, complementing HMG N/O-glycome changes. In conclusion, temporal HMG glycosylation changes provide the groundwork for developing infant formula that is closer to breast milk at different lactation stages.
研究领域:
生物化学
·
营养学
✓ 已验证
DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
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2019
·
IF 6.044
Ying Cai,Wenjiao Yang,Xiaomei Li,Lutan Zhou,Zhongjuan Wang,Lisha Lin,Dingyuan Chen,Longyan Zhao,Zhongkun Li,Shubai Liu,Ronghua Yin,Zhili Zuo,Na Gao,Jinhua Zhao
Fucosylated glycosaminoglycan (FG), a glycosaminoglycan derivative containing distinct sulfated fucose (FucS) branches, displays potent anticoagulant activity by inhibiting the intrinsic tenase complex (iXase). Herein, AmFG, SvFG and HaFG from three species of sea cucumbers were isolated and depolymerized by β-eliminative cleavage. Three series of fragments, A1 - A4 , S1 - S4 and H1 - H4 , were purified from the depolymerized FGs. Based on structural analysis of these fragments, three FGs were deduced as -{→4)-[ L -FucS-α(1→3)]- D -GlcA-β(1→3)- D -GalNAc 4S6S -β(1} n -. The structures differed in sulfation types of FucS , namely, most of FucS in AmFG was Fuc 3S4S , but the FucS in SvFG was Fuc 2S4S , while the FucS in HaFG was Fuc 3S4S , Fuc 2S4S and Fuc 4S . However, all FucS branches attached to C -3 of GlcA as monosaccharides. Anticoagulant and anti-iXase assays showed the octasaccharide is the minimum fragment for potent anticoagulant activity via anti-iXase irrespective of FucS types. Among FG fragments with same degree of polymerization, oligosaccharides containing Fuc 2S4S had more potent anti-iXase activity.
研究领域:
生物化学
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药理学
✓ 已验证
DOI
PubMed
高影响因子
Materials Chemistry Frontiers
·
2016
·Vol. 1
·期 4
·660-667
·
IF 0
Xianfeng Hou,Jin Peng,Fang Zeng,Changmin Yu,Shuizhu Wu
Alpha-L-fucosidase (AFU) plays vital roles in some physiological processes that closely relate to several diseases. Herein, a ratiometric fluorescent probe for the alpha-L-fucosidase (AFU) assay based on a single fluorophore has been developed by functionalizing a new fluorophore platform (6-bromo-anthracenecarboximide) with an α-L-fucose group. AFU cleaves the α-L-fucose group from the probe molecule and remarkably alters its photophysical properties, thus realizing ratiometric detection of AFU. Due to the specificity and high efficiency of the enzymatic reaction, this probe can sensitively and selectively detect AFU in aqueous media and biological milieus with a detection limit of 0.0033 U mL−1. Moreover, the probe can be employed to monitor and spatially map endogenous AFU levels in a hepatocellular carcinoma (HCC) model of zebrafish. Hence, the probe holds significant promise for conducting pathological research on AFU-involved diseases.
研究领域:
生物化学
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医学诊断
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分子生物学
✓ 已验证
DOI
CARBOHYDRATE POLYMERS
·
2025
·
IF 12.5
Chen Xu,Xiang Shi,Huifang Sun,Lijuan Yu,Liang Zhang,Di Lan,Xiaolu Wu,Mengran Chen,Nanqi Cheng,Ying Pan,Jiayi He,Ronghua Yin,Lutan Zhou,Na Gao,Jinhua Zhao
Fucosylated glycosaminoglycan (FG) from Holothuroidea exhibits notable structural diversity and multiple biological activities. This study investigated the HfFG isolated from the sea cucumber Holothuria floridana , focusing on its chemical structure and biological activities. Structural analysis of eleven oligosaccharides (a-k) and a depolymerized product (dHfFG-II) using NMR identified the HfFG backbone as chondroitin sulfate E (CS-E), with various branches, including L-Fuc 2S4S , L-Fuc 3S4S , L-Fuc 4S , and the unique disaccharide D-GalNAc 4S -α1,2-L-Fuc 3S4S , attached at C-3 of GlcA. A high L-Fuc 2S4S (40 %) and disaccharide branch (35 %) content allowed their contiguous distribution within the CS-E chain, as evidenced by the predominant hexasaccharides (i, j) in size-homogeneous Fr4 and the novel nonasaccharide k in Fr5. Notably, previously unreported branches and sequences in HfFG were confirmed, offering new understanding of the natural HfFG structure. HfFG showed potent inhibitory activities on the intrinsic tenase complex (iXase), heparanase, and P-selectin binding to PSGL-1. Depolymerization selectively modulated these activities, preserving anti-iXase potency while attenuating heparanase and P-selectin inhibition. These activities were dependent on oligosaccharide chain length and sequence. Comparing the activities of FG and its oligosaccharides highlights their potential for the rational design of targeted inhibitors of heparanase or P-selectin binding to PSGL-1, with significant implications for therapeutic applications.
研究领域:
海洋生物学
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生物化学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Marine Drugs
·
2019
·Vol. 17
·期 4
·195
·
IF 3.772
Ruowei Guan,Yuan Peng,Lutan Zhou,Wenqi Zheng,Xixi Liu,Pin Wang,Qingxia Yuan,Na Gao,Longyan Zhao,Jinhua Zhao
Apostichopus japonicusis one of the most economically important species in sea cucumber aquaculture in China. Fucosylated glycosaminoglycan fromA. japonicus(AjFG) has shown multiple pharmacological activities. However, results from studies on the structure of AjFG are still controversial. In this study, the deaminative depolymerization method that is glycosidic bond-selective was used to prepare the depolymerized products from AjFG (dAjFG), and then a series of purified oligosaccharide fragments such as tri-, hexa-, nona-, and dodecasaccharides were obtained from dAjFG by gel permeation chromatography. The 1D/2D NMR and ESI-MS spectrometry analyses showed that these oligosaccharides had the structural formula ofl-FucS-α1,3-d-GlcA-β1,3-{d-GalNAc4S6S-β1,4-[l-FucS-α1,3-]d-GlcA-β1,3-}n-d-anTal-diol4S6S(n= 0, 1, 2, 3; FucS represents Fuc2S4S, Fuc3S4S, or Fuc4S). Thus, the unambiguous structure of native AjFG can be rationally deduced: it had the backbone of {-4-d-GlcA-β1,3-d-GalNAc4S6S-β1-}n, which is similar to chondroitin sulfate E, and eachd-GlcA residue in the backbone was branched with al-FucS monosaccharide atO-3. Bioactivity assays confirmed that dAjFG and nonasaccharides and dodecasaccharides from AjFG had potent anticoagulant activity by intrinsic FXase inhibition while avoiding side effects such as FXII activation and platelet aggregation.Keywords:Apostichopus japonicus;fucosylated glycosaminoglycan;oligosaccharide;unambiguous structure;anti-Fxase
研究领域:
海洋生物学
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药理学
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生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2024
·
IF 4.8
Mengshi Xiao,Jiayuan Bi,Xinmiao Ren,Xiaodan Fu,Dongyu Li,Rong Li,Haijin Mou
Fucose-containing oligosaccharides exhibit various biological activities, particularly for improving the intestinal health of infants. Bacterial exopolysaccharides are promising resource of fucose-containing oligosaccharides. A fucose-containing trisaccharide ( d - Glc p-β-(1 → 4)- l - Fuc p and d - Gal p-α-(1 → 3)- l - Fuc p, GuFGa) with same monosaccharide composition with 2′-fucosyllactose (2′-FL, the most abundant oligosaccharide in human milk), is obtained from bacterial exopolysaccharides. This study compared the in vitro prebiotic activities of GuFGa and 2′-FL using mono- and co-culture fermentation. Mono-culture shows GuFGa could be utilized by Bifidobacterium infantis and Lactobacillus plantarum , while 2′-FL only promote the proliferation of B . infantis . Co-culture fermentation involves cross-feeding between B . infantis and L . plantarum to utilize GuFGa. Further in vitro fecal microbiota fermentation of breastfed infants demonstrates that GuFGa exhibits better prebiotic activity than 2′-FL in increasing beneficial bacteria ( Bifidobacterium and Lactobacillus ) abundance and reducing harmful bacteria ( Clostridium _ sensu _ stricto and Escherichia - Shigella ) abundance. Moreover, GuFGa and 2′-FL fermentation shifted the gut microbiota composition of formula-fed infants to that of breastfed infants. Furthermore, herein, GuFGa and 2′-FL caused distinct metabolic profiles in both short-chain fatty acids and indole derivatives (e.g. indolelactic acid and indolepropionic acid). In summary, this study provides insights into the exploration of novel functional fucose-containing oligosaccharides as potential prebiotics.
研究领域:
微生物学
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营养学
·
胃肠病学
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产品关联
DOI
高影响因子
Synthetic and Systems Biotechnology
·
2024
·
IF 4.4
Shanquan Liang,Zi He,Dan Liu,Shaoqing Yang,Qiaojuan Yan,Zhengqiang Jiang
2′-Fucosyllactose (2′-FL) is one of the important functional oligosaccharides in breast milk. So far, few attempts on biosynthesis of 2′-FL by the salvage pathway have been reported. Herein, the salvage pathway enzyme genes were introduced into the E. coli BL21star(DE3) for synthesis of 2′-FL. The 2′-FL titer increased from 1.56 to 2.13 g/L by deleting several endogenous genes on competitive pathways. The α-1,2-fucosyltransferase (WbgL) was selected, and improved the 2′-FL titer to 2.88 g/L. Additionally, the expression level of pathway enzyme genes was tuned through optimizing the plasmid copy number. Furthermore, the spatial distribution of WbgL was enhanced by fusing with the MinD C-tag. After optimizing the fermentation conditions, the 2′-FL titer reached to 7.13 g/L. The final strain produced 59.22 g/L of 2′-FL with 95% molar conversion rate of lactose and 92% molar conversion rate of fucose in a 5 L fermenter. These findings will contribute to construct a highly efficient microbial cell factory to produce 2′-FL or other HMOs.
研究领域:
生物化学
·
微生物学
·
合成生物学
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产品关联
DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY
·
2017
·
IF 4.519
Junhui Li,Shan Li,Lufeng Yan,Tian Ding,Robert J. Linhardt,Yanlei Yu,Xinyue Liu,Donghong Liu,Xingqian Ye,Shiguo Chen
Fucosylated chondroitin sulfates (fCSs) are structurally unusual glycosaminoglycans isolated from sea cucumbers that exhibit potent anticoagulant activity . These fCSs were isolated from sea cucumber, Isostichopus badionotus and Pearsonothuria graeffei . Fenton reaction followed by gel filtration chromatography afforded fCS oligosaccharides , with different sulfation patterns identified by mass and NMR spectroscopy , and these were used to clarify the relationship between the structures and the anticoagulant activities of fCSs. In vitro activities were measured by activated partial thromboplastin time (APTT), thrombin time (TT), thrombin and factor Xa inhibition, and activation of FXII . The results showed that free radicals preferentially acted on GlcA residues affording oligosaccharides that were purified from both fCSs. The inhibition of thrombin and factor X activities, mediated through antithrombin III and heparin cofactor II of fCSs oligosaccharides were affected by their molecular weight and fucose branches. Oligosaccharides with different sulfation patterns of the fucose branching had a similar ability to inhibit the FXa by the intrinsic factor Xase (factor IXa-VIIIa complex). Oligosaccharides with 2,4- O -sulfo fucose branches from fCS- Ib showed higher activities than ones with 3,4- O -disulfo branches obtained from fCS- Pg . Furthermore, a heptasaccharide is the minimum size oligosaccharide required for anticoagulation and FXII activation. This activity was absent for fCS oligosaccharides smaller than nonasaccharides . Molecular size and fucose branch sulfation are important for anticoagulant activity and reduction of size can reverse the activation of FXII caused by native fCSs.
研究领域:
生物化学
·
药理学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）