Scientific Reports
·
2015
·Vol. 5
·期 1
·1-12
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IF 5.578
Yang Yang,Zhang Xinxin,Yin Qiang,Fang Wei,Fang Zemin,Wang Xiaotang,Zhang Xuecheng,Xiao Yazhong
β-Glucosidases are enzymes that hydrolyze β-glycosidic bonds to release non-reducing terminal glucosyl residues from glycosides and oligosaccharides and thus have significant application potential in industries. However, most β-glucosidases are feedback inhibited by the glucose product, which restricts their application. Remarkably, some β-glucosidases of the glycoside hydrolase (GH) 1 family are tolerant to or even stimulated by glucose. Elucidation of the mechanisms of glucose tolerance and stimulation of the GH1 β-glucosidases will be crucial to improve their application through enzyme engineering. In this study, by comparing the primary and tertiary structures of two GH1 β-glucosidases with distinct glucose dependence, some putative glucose-dependence relevant sites were mutated to investigate their exact roles. Both biochemical and structural characterization of the mutants suggested that some sites at the entrance and middle of the substrate channel regulate the effects of glucose and the relative binding affinity/preference of these sites to glucose modulates the glucose dependence. A mechanism was therefore proposed to interpret the glucose dependence of GH1 β-glucosidases. This research provides fresh insight into our current understanding of the properties and mechanisms of GH1 β-glycosidases and related enzymes that modulate their activity via feedback control mechanism.
研究领域:
生物化学
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酶学
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生物技术
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2018
·Vol. 410
·期 7
·2011-2018
·
IF 3.307
Yu Lingling,Wen Chao,Li Xing,Fang Shiqi,Yang Lichuan,Wang Tony,Hu Kaifeng
Quantification of endogenous and exogenous plasma glucose can help more comprehensively evaluate the glucose metabolic status. A ratio-based approach using isotope dilution liquid chromatography tandem mass spectrometry (ID LC-MS/MS) with indirect multiple reaction monitoring (MRM) of the derivative tag was developed to simultaneously quantify endo-/exogenous plasma glucose. Using diluted D-[ 13 C 6 ] glucose as tracer of exogenous glucose, 12 C 6 / 13 C 6 glucoses were first derivatized and then data were acquired in MRM mode. The metabolism of exogenous glucose can be tracked and the concentration ratio of endo/exo-genous glucose can be measured by calculating the endo-/exo-genous glucose concentrations from peak area ratio of specific daughter ions. Joint application of selective derivatization and MRM analysis not only improves the sensitivity but also minimizes the interference from the background of plasma, which warrants the accuracy and reproducibility. Good agreement between the theoretical and calculated concentration ratios was obtained with a linear correlation coefficient ( R ) of 0.9969 in the range of D-glucose from 0.5 to 20.0 mM, which covers the healthy and diabetic physiological scenarios. Satisfactory reproducibility was obtained by evaluation of the intra- and inter-day precisions with relative standard deviations (RSDs) less than 5.16%, and relative recoveries of 85.96 to 95.92% were obtained at low, medium, and high concentration, respectively. The method was successfully applied to simultaneous determination of the endo-/exogenous glucose concentration in plasma of non-diabetic and type II diabetic cynomolgus monkeys.
Graphical Abstract The scheme of the proposed ratio-based approach using isotope dilution LC-MS/MS with indirect MRM of the derivative tag for simultaneous quantification of endogenous and exogenous plasma glucose.
研究领域:
分析化学
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代谢组学
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生物医学科学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
·
2021
·
IF 8.128
Huijuan Yang,ShengBao Wang,Xingpu Wang,Pengyang Zhang,Cheng Yan,Yangyang Luo,Lina Chen,Mengjiao Li,Fan Fan,Zhiyou Zhou,Xifei Li
Glucose oxidation reaction (GOR) plays a significant role in glucose fuel cells anode and glucose sensors. Therefore, optimizing the GOR catalyst nanostructure is auxiliary to their efficient operation. In this study, we present a cascade-assembled strategy to prepare CuO nanobundles (CuO-NB) with high-density and homogenous grain boundaries (GBs). The essence of activity in GOR that depended on GBs are thoroughly investigated. The increased glucose diffusion coefficient of CuO-NB means that GBs has a faster glucose mass transfer, which is attributed to the terraces in GBs dislocation surface. Furthermore, the accumulation of electrons on GBs makes the glucose adsorption increased and the free energy of dehydrogenation step decreased, leading to a lower glucose oxidation barrier. Therefore, CuO-NB is appropriate for non-invasive glucose detection and glucose fuel cells. This study sheds new light on the GBs effect in GOR and paves the way for developing high-efficiency electrocatalysts.
研究领域:
材料科学
·
电化学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2025
·Vol. 26
·期 7
·3118
·
IF 4.9
Guotao Mao,Ming Song,Hao Li,Junhan Lin,Kai Wang,Qian Liu,Zengping Su,Hongsen Zhang,Lijuan Su,Hui Xie,Andong Song
The enzymatic hydrolysis of lignocellulose is often hindered by the glucose-mediated inhibition of β-glucosidases, a major bottleneck in industrial cellulose degradation. Identifying novel glucose-tolerant β-glucosidases is essential for enhancing saccharification efficiency. In this study, we cloned and heterologously expressed a novel β-glucosidase, RpBgl8, from the termiteReticulitermes perilucifugusinEscherichia coli. Sequence and structural analyses classified RpBgl8 as a glycoside hydrolase family 1 enzyme. The purified enzyme exhibited optimal activity at 45 °C and pH 7.0, with broad stability across pH 4.0–8.0. Notably, RpBgl8 demonstrated high tolerance to lignocellulose-derived inhibitors and organic solvents, maintaining 100% activity in 15% ethanol. Furthermore, RpBgl8 exhibited outstanding glucose tolerance, retaining 100% activity at 2.5 M glucose and 82% activity at 4.0 M glucose—outperforming most previously reported β-glucosidases. A structural analysis revealed a narrow, hydrophobic substrate pocket, with residue F124 at the glycone-binding site critical for minimizing glucose accumulation. The F124W mutation significantly reduced glucose tolerance, confirming that hydrophobic interactions at the active site mitigate inhibition. These findings establish RpBgl8 as a promising candidate for high-solid biomass processing and simultaneous saccharification and fermentation applications, highlighting termites as underexplored sources of biocatalysts with unique industrial potential.
研究领域:
生物化学
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工业酶学
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结构生物学
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产品关联
DOI
PubMed
高影响因子
Microbial Cell Factories
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2022
·Vol. 21
·期 1
·1-14
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IF 6.352
Guo Fei,Zhang Xingwang,You Cai,Zhang Chengjie,Li Fengwei,Li Nan,Xia Yuwei,Liu Mingyu,Qiu Zetian,Zheng Xianliang,Ma Li,Zhang Gang,Luo Lianzhong,Cao Fei,Feng Yingang,Zhao Guang-Rong,Zhang Wei,Li Shengying,Du Lei
Background Glucoside natural products have been showing great medicinal values and potentials. However, the production of glucosides by plant extraction, chemical synthesis, and traditional biotransformation is insufficient to meet the fast-growing pharmaceutical demands. Microbial synthetic biology offers promising strategies for synthesis and diversification of plant glycosides.Results In this study, the two efficient UDP-glucosyltransferases (UGTs) (UGT85A1 and RrUGT3) of plant origin, that are capable of recognizing phenolic aglycons, are characterized in vitro. The two UGTs show complementary regioselectivity towards the alcoholic and phenolic hydroxyl groups on phenolic substrates. By combining a developed alkylphenol bio-oxidation system and these UGTs, twenty-four phenolic glucosides are enzymatically synthesized from readily accessible alkylphenol substrates. Based on the bio-oxidation and glycosylation systems, a number of microbial cell factories are constructed and applied to biotransformation, giving rise to a variety of plant and plant-like O-glucosides. Remarkably, several unnatural O-glucosides prepared by the two UGTs demonstrate better prolyl endopeptidase inhibitory and/or anti-inflammatory activities than those of the clinically used glucosidic drugs including gastrodin, salidroside and helicid. Furthermore, the two UGTs are also able to catalyze the formation of N- and S-glucosidic bonds to produce N- and S-glucosides.Conclusion sTwo highly efficient UGTs, UGT85A1 and RrUGT3, with distinct regioselectivity were characterized in this study. A group of plant and plant-like glucosides were efficiently synthesized by cell-based biotransformation using a developed alkylphenol bio-oxidation system and these two UGTs. Many of the O-glucosides exhibited better PEP inhibitory or anti-inflammatory activities than plant-origin glucoside drugs, showing significant potentials for new glucosidic drug development.
研究领域:
药学
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生物技术
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合成生物学
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生物化学
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药物化学
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DOI
PubMed
高影响因子
JOURNAL OF APPLIED TOXICOLOGY
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2018
·Vol. 38
·期 6
·810-823
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IF 2.909
Zhangjian Chen,Yun Wang,Xiang Wang,Lin Zhuo,Shi Chen,Shichuan Tang,Lin Zhao,Xianguo Luan,Guang Jia
As food additives, titanium dioxide nanoparticles (TiO 2 NPs) have been widely used in various products that are usually simultaneously consumed with a high content of sugar, thus necessitating research on the effect of TiO 2 NPs on glucose homeostasis. We conducted an animal study to explore the effect of orally administrated TiO 2 NPs on glucose absorption and metabolism in rats at 0, 2, 10 and 50 mg kg –1 body weight day –1 for 30 and 90 days. The results showed that oral exposure to TiO 2 NPs caused a slight and temporary hypoglycemic effect in rats at 30 days post-exposure but recovered at 90 days post-exposure. Decreased levels of intestinal glucose absorption and increased levels of hepatic glucose metabolism may be responsible for the hypoglycemic effect. Remodeling of the villi in the small intestine that decreased the surface area available for glucose absorption and increased levels of hepatic glucose uptake, utilization and storage related to hepatocellular injury are supposed to be the mechanisms. Our results demonstrated that dietary intake of TiO 2 NPs as food additives could affect the absorption and metabolism of glucose.
研究领域:
食品科学
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毒理学
·
代谢
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2012
·
IF 4.98
Zheng-Zheng Zou,Hui-Lei Yu,Chun-Xiu Li,Xin-Wen Zhou,Chihiro Hayashi,Jie Sun,Bao-Hong Liu,Tadeyuki Imanaka,Jian-He Xu
A new β-glucosidase (DtGH) representing 40% identity with an apple seed glycosidase (ASG) was cloned from Dictyoglomus thermophilum . DtGH showed extremely high thermostability in aqueous solution, with half-lives of 533, 44, and 5 h measured at 70, 80 and 90 °C, respectively. Therefore it was used for direct glycosylation of n -octanol at 70 °C instead of 50 °C as usually. As a result, the glucose based conversion was increased by 27%, but the time spent to reach equilibrium was decreased from 7 d to 3 d. This enzyme also exhibited excellent stability under the reaction environment, retaining 70–80% of its initial activity after 7 d of incubation at 70 °C in either 1.7 M glucose solution or octanol-aqueous (85:15, v/v) system. It could retain part of synthetic activity even in boiling water. Owing to the strong glucose-tolerance and extremely high thermostability, DtGH should be promising for various glucosides synthesis.
研究领域:
生物化学
·
酶工程
✓ 已验证
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
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2025
·
IF 5.4
Aleksandr L. Kim,Egor V. Musin,Sergey A. Tikhonenko
Enzymatic analysis methods are widely used for the determination of low-molecular-weight substances in various fields; however, they have several limitations. One promising approach to addressing these limitations is the encapsulation of enzymes in polyelectrolyte microcapsules (PMCs). In this study, the development of a bienzymatic diagnostic system based on two separately encapsulated enzymes—urease and glucose oxidase (GO)—is proposed for the first time, enabling the simultaneous determination of urea and glucose concentrations in a single sample. To achieve this, the influence of reaction products and substrates of urease on the activity of GO, and vice versa, was investigated. It was demonstrated that ammonia reduces the activity of glucose oxidase, while gluconic acid and hydrogen peroxide decrease the activity of urease, with the PMC shell exhibiting protective properties for the encapsulated form of urease. Other reaction components did not affect the enzyme activities. The linear range, coefficient of variation, and reusability of this diagnostic system were determined: the linear range for urea concentration detection was found to be between 10 and 50 mM (coefficient of variation 2 %) with 7 reuses of same PMC-urease; the linear range for glucose concentration detection was between 30 and 100 mM (coefficient of variation 8 %) with 13 reuses of same PMC-GO. This diagnostic system is suitable for determining urea and glucose concentrations in industrial biofuel production and in industrial fermenters using urea as a nitrogen source.
研究领域:
生物化学
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诊断学
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产品关联
DOI
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2025
·
IF 2.5
Hongbing Mao,Huijun Wang,Yanyan Xing,Deyong Wang,Xiujun Cao,Jie Yu,Junya Yan
In this work, a boronic acid decorated mesoporous iron-based metal-organic framework (MIL-100(Fe)-B) nanozyme as an efficient immobilization matrix for glucose oxidase (GOx) was designed and prepared through a facile microwave-assisted metal-ligand-fragment co-assembly strategy. Compared with the unboric acid-modified MIL-100(Fe), boron-doped MIL-100(Fe) exhibited enhanced peroxidase-like activity, which was mainly due to the introduced B causing MIL-100(Fe) defects, accelerating mass transfer, facilitating more active sites for H2O2 decomposition. Besides, the MIL-100(Fe)-B nanozyme possessed sufficient recognition sites for immobilizing glucose oxidase with little enzyme leakage to form a cascade nanozyme with both peroxidase-like and cascade catalytic activities. A colormetric biosensor based on the integrated nanozyme reactor (GOx@MIL-100(Fe)-B) was developed for glucose detection. The proposed cascade biosensor had a linear range of 5-150 μM and a limit of detection (LOD) of 0.97 μM (3.3 δ/S). Interestingly, a portable microfluidic paper-based analytic device (μPAD) biosensor based on the cascade nanoplatform was further fabricated for visualized detection of glucose combined with a ColorPicker App on smartphone. It indicated that the integrated nanozyme GOx@MIL-100(Fe)-B was a promising candidate for glucose detection and provided a potentially analytical avenue for biosensing and other applications.
研究领域:
生物化学
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分析化学
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材料科学
✓ 已验证
产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 6
·1-7
·
IF 5.479
Qiao Juan,Liu Qianrong,Wu Han,Cai Huiwu,Qi Li
A fluorescent probe is described for the determination of serum glucose after hepatotoxin-induced liver injury. The probe is based on the use of a water-soluble polymer and has been prepared from a multi-functional azlactone polymer as the linker, amino boronic acid, and Alizarin Red as the signalling moiety. The excitation/emission peaks of the polymeric fluorescent probe are at 468/567 nm. Fluorescence is reduced on addition of glucose. Intensity drops linearly in the 0.1 mM to 14 mM glucose concentration range. The probe was applied to non-enzymatic detection of glucose in rat serum after CCl 4 -induced liver damage.
A polymer based fluorescent probe has been constructed and applied for non-enzymatic monitoring of serum glucose following hepatotoxin induced liver injury.
研究领域:
生物化学
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医学诊断
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材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2017
·
IF 3.154
Liuxi Xia,Yuxiang Bai,Wanmeng Mu,Jinpeng Wang,Xueming Xu,Zhengyu Jin
Instead of β-cyclodextrin (β-CD), branched β-CDs have been increasingly used in many aspects as they possess better solubility and higher bioadaptability. But most commercialized branched β-CDs were chemically synthesized. Thus, the glucosyl-β-cyclodextrin (G1-β-CD) prepared via enzymatic approach could be a nice substitute. However, the yield of G1-β-CD was low. Here, we reported a controlled two-step reaction to efficiently prepare G1-β-CD from maltodextrins by β-cyclodextrin glucosyltransferase (β-CGTase) and amyloglucosidase (AG). Compared to the single β-CGTase reaction, controlled two-step reaction caused a yield increase of G1-β-CD by 130%. Additionally, the percentage of G1-β-CD was enhanced from 2.4% to 24.0% and the side products α-CD and γ-CD were hydrolyzed because of the coupling activity of β-CGTase. Thus, this controlled two-step reaction might be an efficient approach for industrial production of pure G1-β-CD.
研究领域:
生物化学
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食品科学
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2020
·
IF 10.257
Zhen Liang,Jieyu Zhang,Can Wu,Xuefeng Hu,Yuhui Lu,Guangfeng Wang,Fei Yu,Xiaojun Zhang,Yunbing Wang
The commercial sensor of a continuous glucose monitoring system suffers from restricted penetration of glucose in the dense sensing coating, uncontrolled leakage of the glucose oxidase and electrocatalytic medium, and susceptibility to mechanical damage. Herein, a self-healing hydrogel based on quaternized chitosan and oxidized dextran was designed, and CeO 2 /MnO 2 hollow nanospheres were covalently linked in the hydrogel as the electrocatalytic medium. Glucose oxidase was loaded via the strong electrostatic interactions with the CeO 2 /MnO 2 hollow nanospheres. An extra covering agent was coated on the hydrogel to prevent the leakage of the glucose oxidase and electrocatalytic medium. Covalent linkage of the hydrogel on a bendable chip formed a flexible glucose sensor , which showed a wide linear range (1–111 mM), fast response (less than 3 s), and high sensitivity (176 μA mM −1 cm −2 ). The hydrogel-based sensor was self-healable, and could continuously work for over 30 days. Thus, this study provides a method to simultaneously prevent the leakage of the electrocatalytic medium, promote the sensitivity of glucose detection, and tolerate the mechanical damage, which shows great potential for continuous glucose monitoring.
研究领域:
生物医学工程
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材料科学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）