BIORESOURCE TECHNOLOGY
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2025
·
IF 9
Jiaqi Ding,Xiaoxiao Li,Zhengyu Jin,Birte Svensson,Yuxiang Bai
CD complexation solubilization and enzymatic glycosylation synergistically enhance the utilization of polyphenols in biomass. In this study, a comprehensive strategy was proposed, to regulate the rate-limiting step where the efficiency of β -CD ring-opening ( k 1 ) exceeded the daidzein glycosylation ( k 2 ) catalyzed by CGTase in the efficient glycosylation system based on cyclodextrin (CD) dynamic complexation. This approach involved systematic regulation of the CD:polyphenol molar ratio and concentration, along with rational CGTase engineering, to enhance the glycosylation yield of polyphenols. Through molecular docking and sequence alignment, key residues Y195 and E264 near the acceptor subsites +1 and +3 were identified. Based on the principles of enhancing hydrophobicity and reducing steric hindrance, site-directed mutagenesis was performed on the two residues, and screening obtained seven single mutants with improved polyphenol glycosylation efficiency. The half-reaction kinetics analysis showed that mutants Y195F, E264A and E264V successfully narrowed the gap between k 1 and k 2 . Finally, the double mutants Y195F/E264A and Y195F/E264V were constructed, which showed higher glycosylation yields than single mutants. The optimal mutant Y195F/E264V achieved the highest reported yields of daidzein 7-O- α - d -glucopyranoside (65.6 %) and rutin 4″-O- α - d -glucopyranoside (89.4 %) in water, surpassing the wild-type (WT) by 37.4 % and 8.3 %, respectively. The study presents a universal approach for the valorization of plant biomass, with the potential for scalable application in plant polyphenol utilization.
研究领域:
生物化学
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食品科学
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植物学
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产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2025
·
IF 9
Jingwen Zhang,Binge Zhang,Feiyu Duan,Zehui Xuan,Tong Sun,Lili Lu
β-Galactosidases are important enzymatic tools for glycosylation, but their properties vary greatly with the source. Here, ten putative β-galactosidase genes, designated as bga 1 to bga 10, encoding proteins Bga1 to Bga10, were mined from an environmental metagenomic dataset comprising 119,152 sequences. Five of the encoded enzyme proteins exhibited less than 80% sequence similarity to known enzymes, but displayed conserved catalytic sites in their predicted three-dimensional models. After heterologous expression and characterization, two recombinant enzymes showed specific hydrolysis activity toward o -nitrophenyl-β- d -galactopyranoside. One of them, Bga4R, exhibited remarkable activity at pH 7.4 and 50℃, with excellent alkaline stability. Notably, Bga4R tolerated a wide range of acceptors for transglycosylation. It catalyzed galactosyl transfer to various monosaccharides and sugar alcohols, and enabling the synthesis of diverse glycosylated derivatives. This study identifies a novel GH 1 β-galactosidase as a powerful tool for glycosylation engineering, with promising potential for synthesizing galactosides valuable to food and pharmaceutical industries.
研究领域:
生物化学
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微生物学
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合成生物学
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2016
·Vol. 6
·期 1
·1-10
·
IF 5.228
Liang Chaoning,Zhang Yi,Jia Yan,Wenzhao Wang,Li Youhai,Lu Shikun,Jin Jian-Ming,Tang Shuang-Yan
Glycodiversification broadens the scope of natural product-derived drug discovery. The acceptor substrate promiscuity of glucosyltransferase-D (GTF-D), a carbohydrate-processing enzyme from Streptococcus mutans , was expanded by protein engineering. Mutants in a site-saturation mutagenesis library were screened on the fluorescent substrate 4-methylumbelliferone to identify derivatives with improved transglycosylation efficiency. In comparison to the wild-type GTF-D enzyme, mutant M4 exhibited increased transglycosylation capabilities on flavonoid substrates including catechin, genistein, daidzein and silybin, using the glucosyl donor sucrose. This study demonstrated the feasibility of developing natural product glycosyltransferases by engineering transglycosidases that use donor substrates cheaper than NDP-sugars, and gave rise to a series of α-glucosylated natural products that are novel to the natural product reservoir. The solubility of the α-glucoside of genistein and the anti-oxidant capability of the α-glucoside of catechin were also studied.
研究领域:
生物化学
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药物开发
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合成生物学
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DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2025
·
IF 3.7
Chen-Xin Guo,Jiyan Li,Yu Wang,Tang Li,Heng Yin
Stevia rebaudiana , a perennial herb, is recognized not only for its sweet steviol glycosides but also for its rich flavonoid content, which confer pharmacological properties including anti-inflammatory, antimicrobial, and anticancer activities. However, the enzymatic basis underlying flavonoid modification in S. rebaudiana remains poorly understood. In this study, we identified, cloned, and heterologously expressed a novel flavonoid glycosyltransferase gene, SrUGT72B1 in E. coli . The recombinant SrUGT72B1 catalyzed the glycosylation of multiple flavonoids using UDP-glucose as the primary sugar donor, and exhibited broad substrate promiscuity toward apigenin, luteolin, phloretin and kaempferol. In addition to UDP-glucose, SrUGT72B1 also accepted UDP-xylose and UDP-rhamnose, with UDP-glucose exhibiting the highest catalytic efficiency. Biochemical characterization revealed that the enzyme functions optimally at pH 9.0 and 50 °C. Notably, SrUGT72B1 demonstrates regioselective 5-O-glycosylation toward apigenin, a rare activity among plant glycosyltransferases. Molecular docking and molecular dynamics simulations provided structural insights into this unique regioselectivity and substrate recognition. Together, these findings establish SrUGT72B1 as a previously uncharacterized flavonoid 5-O-glycosyltransferase, expanding the functional landscape of plant UGTs and offering potential applications in the biosynthesis of value-added flavonoid glycosides.
研究领域:
植物学
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生物化学
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分子生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
PLANT PHYSIOLOGY AND BIOCHEMISTRY
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2025
·
IF 5.7
Jiafei Qian,Chuanhong Ren,Fan Wang,Yunlin Cao,Yan Guo,Xiaoyong Zhao,Yilong Liu,Changqing Zhu,Xiaoying Li,Hongxia Xu,Junwei Chen,Kunsong Chen,Xian Li
Flavonol glycosides are secondary metabolites important for plant development and stress defense such as UV-B irradiation. UDP-glycosyltransferase (UGT) is the last enzyme step that catalyzes flavonol glycosides biosynthesis. Eriobotrya japonica is abundant in flavonol glycosides, but UGTs responsible for accumulation of flavonol glycosides remain unknown. Here, 13 flavonol glycosides including monoglycosides and diglycosides were characterized in different tissues of loquat by LC-MS/MS. UV-B irradiation significantly increased the accumulation of four quercetin glycosides and two kaempferol glycosides in loquat fruit. Based on UGT gene family analysis, transcriptome analysis, enzyme assays of recombinant proteins as well as transient overexpression assays in Nicotiana benthamiana , three UGTs were identified, i.e. EjUGT78T4 as flavonol 3- O -galactosyltransferase, EjUGT78S3 as flavonol 3- O -glucosyltransferase, and EjUGT91AK7 as flavonol 1→6 rhamnosyltransferase. This work elucidates the formation of flavonol glycosides in loquat through UGT-mediated glycosylation.
研究领域:
植物学
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分子生物学
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代谢组学
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产品关联
DOI
PubMed
高影响因子
AICHE JOURNAL
·
2024
·Vol. 70
·期 9
·e18512
·
IF 3.5
Hanchi Chen,Lingjun Ju,Yangyang Dong,Shijie Lu,Yingling Bao,Linjiang Zhu,Xiaolong Chen
Cyclodextrin glycosyltransferase (CGTase) catalyzes intermolecular transglycosylation through either disproportionation or cyclization-coupling pathway. Kinetics analysis reveals that the hesperidin glycosylation process catalyzed by a CGTase variant (M1) is primarily accomplished through the disproportionation pathway. The cyclization-coupling pathway exhibits a lower reaction rate and competitively consumes glycosyl donor and yield byproducts that impair disproportionation. Under the guidance of reaction kinetics, mutagenesis was targeted at residues in the −3, +1, and +2 subsites, known to control the selectivity between disproportionation and cyclization. A quadruple variant was identified with 2.9 times hesperidin glycosylation activity compared to M1, and 20.3 times compared to the wild-type. Kinetic analysis reveals a fourfold improvement of k cat / K mA for disproportionation and an 85.5% reduction in k cat / K m for cyclization after mutagenesis. Binding free energy analysis further confirms that the mutagenesis favors the binding of hesperidin, and destabilizes the binding of cyclodextrin.
研究领域:
生物化学
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酶工程
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产品关联
DOI
高影响因子
RSC Advances
·
2018
·Vol. 8
·期 63
·35870-35877
·
IF 2.936
Guoying Su,Lin Li,Di Zhao,Bing Li,Xia Zhang
The digestion of dietary advanced glycation end products (AGEs) largely determines their absorption in humans. To help elucidate the health effects of dietary AGEs, changes in the digestive behavior of bovine serum albumin (BSA, dietary protein) caused by glycation derived from glyoxal (GO, an important precursor of AGEs) in a simulated food heating system have been investigated. The hydrothermal aggregation of BSA was suppressed by GO derived glycation, generating glycated aggregates of loose and branched structures, according to dynamic light scattering (DLS), circular dichroism (CD) spectroscopy, free sulfhydryl group, transmission electron microscopy (TEM) and small angle X-ray scattering (SAXS) results. Analysis of protein digests showed that glycation reduced the gastric and gastrointestinal digestibility of BSA and the bioavailability of all seven detected amino acids. A comparative analysis of the distribution of CML and lysine in glycated BSA digests with different molecular weights showed that carboxymethylation directly blocked the action of proteases on Lys residues.
研究领域:
食品科学
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营养学
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生物化学
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产品关联
DOI
PubMed
高影响因子
PLANT JOURNAL
·
2021
·Vol. 106
·期 2
·351-365
·
IF 6.486
Cheng-Yong Feng,Shan-Shan Li,Goro Taguchi,Qian Wu,Dan-Dan Yin,Zhao-Yu Gu,Jie Wu,Wen-Zhong Xu,Cheng Liu,Liang-Sheng Wang
SUMMARY Lotus plumule, the embryo of the seed of the sacred lotus ( Nelumbo nucifera ), contains a high accumulation of secondary metabolites including flavonoids and possesses important pharmaceutical value. Flavonoid C -glycosides, which accumulate exclusively in lotus plumule, have attracted considerable attention in recent decades due to their unique chemical structure and special bioactivities. As well as mono- C -glycosides, lotus plumule also accumulates various kinds of di- C -glycosides by mechanisms which are as yet unclear. In this study we identified two C -glycosyltransferase (CGT) genes by mining sacred lotus genome data and provide in vitro and in planta evidence that these two enzymes (NnCGT1 and NnCGT2, also designated as UGT708N1 and UGT708N2, respectively) exhibit CGT activity. Recombinant UGT708N1 and UGT708N2 can C -glycosylate 2-hydroxyflavanones and 2-hydroxynaringenin C -glucoside, forming flavone mono- C -glycosides and di- C -glycosides, respectively, after dehydration. In addition, the above reactions were successfully catalysed by cell-free extracts from tobacco leaves transiently expressing NnCGT1 or NnCGT2. Finally, enzyme assays using cell-free extracts of lotus plumule suggested that flavone di- C -glycosides (vicenin-1, vicenin-3, schaftoside and isoschaftoside) are biosynthesized through sequentially C -glucosylating and C -arabinosylating/ C -xylosylating 2-hydroxynaringenin. Taken together, our results provide novel insights into the biosynthesis of flavonoid di- C -glycosides by proposing a new biosynthetic pathway for flavone C -glycosides in N. nucifera and identifying a novel uridine diphosphate-glycosyltransferase (UGT708N2) that specifically catalyses the second glycsosylation, C -arabinosylating and C -xylosylating 2-hydroxynaringenin C -glucoside.
研究领域:
植物生物化学
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分子生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
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2024
·
IF 6
He Li,Dan Wang,Chen-Yu Liu,Hu-Tan-Xian Zhang,Feng-Ru Lv,Jia-Tong He,Zheng-Yang Zhou,Tian-Yu Hou,Hui-Zhen Li
Protein glycosylation is an effective way to enhance the properties of proteins in food systems. However, the negative effects of glycosylation, such as generation of advanced glycation end products (AGEs), have received less attention; these effects are closely related to the occurrence of various chronic metabolic diseases. In this study, a perilla seed meal protein (PSMP)-fructose/dextran (DEX)-caffeic acid (CA) ternary emulsifying system was constructed. Then, AGEs formation and emulsifying properties were characterized, and the optimal emulsifying system for β-carotene (BC) delivery was investigated. The resulting complex was examined using a high-performance liquid chromatography (HPLC)-ultraviolet (UV) detector, ultra-HPLC-tandem mass spectrometry (UHPLC-MS/MS) system, and nanoparticle size analyzer. The results indicated that CA conjugated with glycosylated PSMP significantly reduced the formation of fructosamine (13.5%), methylglyoxal (33.26%), N ε -carboxymethyl lysine, and N ε -carboxyethyl lysine (>80%). Compared with other conjugates, the PSMP-DEX-CA ternary conjugate showed the lowest AGEs content, smaller particle size, better emulsifying active index, lower polydispersity index, and acceptable ζ-potential. The emulsifying system constructed using abovementioned ternary conjugate revealed better UV and thermal stability in BC delivery, which slowed BC degradation. This study provides a novel approach for constructing a low AGEs emulsion system to deliver BC via PSMP with different modifications.
研究领域:
食品科学
·
营养学
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产品关联
DOI
高影响因子
Catalysts
·
2019
·Vol. 9
·期 9
·734
·
IF 3.444
Jian Zhang,Yan Shao,Weiwei Wang
Curcumin is a naturally occurring polyphenolic compound that is commonly used in both medicine and food additives, but its low aqueous solubility and poor bioavailability hinder further clinical applications. For assessing the effect of the glycosylation of curcumin on its aqueous solubility, two glycosyltransferase genes (BsGT1andBsGT2) were cloned from the genome of the strainBacillus subtilisATCC 6633 and over-expressed inEscherichia coli.Then, the two glycosyltransferases were purified, and their glycosylation capacity toward curcumin and its two analogues was verified. The results showed that bothBsGT1andBsGT2could convert curcumin and its two analogues into their glucosidic derivatives. Then, the structures of the derivatives were characterized as curcumin 4′-O-β-D-glucoside and two new curcumin analogue monoglucosides namely, curcumoid-O-α-D-glucoside (2a) and 3-pentadienone-O-α-D-glucoside (3a) by nuclear magnetic resonance (NMR) spectroscopy. Subsequently, the dissolvability of curcumin 4′-O-β-D-glucoside was measured to be 18.78 mg/L, while its aglycone could not be determined. Furthermore, the optimal catalyzing conditions and kinetic parameters ofBsGT1andBsGT2toward curcumin were determined, which showed that the Kcat value ofBsGT1was about 2.6-fold higher than that ofBsGT2, indicating that curcumin is more favored forBsGT2. Our findings effectively apply the enzymatic approach to obtain glucoside derivatives with enhanced solubility.Keywords:curcumin 4′-O-β-D-glucoside;glycosyltransferases;Bacillus subtilisATCC 6633
研究领域:
生物化学
·
酶学
·
药理学
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产品关联
DOI
高影响因子
ACS Catalysis
·
2025
·
IF 13.1
Zhuqing Wang,Jiahui Li,Xueyun Wang,Boyu Jin,Liwei Zhou,Zili Zhao,Meijia Gu,Xuemin Song,Jiahong Wang,Zixin Deng,Shuwen Wu,Zhengyu Zhang,Wenqing Chen
3′-O-β-Glucosyl purine-related nucleosides are actinobacterial natural products with intricate structures, in which the glucosyl attachment to a nucleoside scaffold is governed by a glycosyltransferase. However, the molecular logic and engineered application of the glycosyltransferase have nearly remained unexplored. Here, we report the discovery, characterization, and exploitation of the thermophilic glycosyltransferase ScaGT. We uncover that ScaGT and its homologue AvpGT indicate prominent promiscuity against both sugar donors and a variety of nucleosides. Remarkably, we have solved the ternary complex structure of AvpGT, unveiling that it employs an unpreceded “twin-tyrosine gate” mechanism for substrate recognition and promiscuity, and we have also realized directed biosynthesis of diversified purine nucleoside analogues with unexpectedly enhanced titer via introduction of the external scaGT or avpGT. Moreover, we reveal that 3′-O-β-glucosyl ribavirin exhibits significantly enhanced antiviral activities, thereof showing its promising application potentials. Finally, we have further achieved the gram-scale production of 3′-O-β-glucosyl ribavirin by both biocatalytic and fermentation-feeding strategies. These findings expand the biochemical repertoire regarding glycosyltransferase reactions and provide the basis for rapid mining and rational engineering of more related glycosyltransferases toward synthetic biology applications.
研究领域:
生物化学
·
合成生物学
·
酶学
·
抗病毒药物开发
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产品关联
DOI
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2022
·
IF 3.322
Zhiming Wang,Yuedong He,Lijing Liao,Yuhao Zhang,Yucheng Zhao,Yibei Xiao,Xuefei Jiang,Fei Qiao
A glycosyl transferase , isolated from Angelica decursiva a medical herb rich in coumarin , shows C-glycosyl transferase activity by in vitro activity assay using 5,7-dihydroxycoumarin as substrate, producing a C-glycosylated product at position C′8 along with the main product at C′6 position. Catalytic promiscuity assay shows that Ad CGT also displays O- or C-glycosylation activity to other coumarins and flavonoids . When phloretin and 2,4,6-trihydroxyacetophenone were fed as substrates, Ad CGT catalyzed the formation of di-C-glycosides. Therefore, Ad CGT is a multifunctional glycosyltransferase with a broad substrate acceptability. This work highlights the potential of Ad CGT as a catalyst for glycosylation of coumarin and reveals a new regio-selective C-glycosyltransferase, providing a basis for exploring the mechanism of coumarin glycosylation.
研究领域:
生物化学
·
生物催化
·
药物化学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）