INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2017
·
IF 3.671
Xiuting Hu,Yu Wang,Chengmei Liu,Zhengyu Jin,Yaoqi Tian
Uricase was conjugated with dextrin to improve its stability. Firstly, dextrin was succinylated into dextrin monosuccinate with the functional group, carboxylic acid, and dextrin monosuccinates with different degree of substitution (DS) were prepared. Secondly, dextrin-uricase conjugate was synthesized by uricase and dextrin monosuccinate, which was verified by size-exclusion chromatography and anion exchange chromatography . Thirdly, it was found that the conjugate degree of the dextrin-uricase conjugate was positively related to the DS of dextrin monosuccinate, but the activity was seriously lost and difficult to recover by α-amylase. When the molar ratio of dextrin monosuccinate (DS = 0.283) to uricase was 30:1, the conjugate degree of dextrin-uricase conjugate reached 22.1% and the activity was decreased to 40.4%. After triggered by α-amylase, the activity was recovered to 83.4%. Finally, the enzymatic properties of this dextrin-uricase conjugate were investigated and compared with those of free uricase. The optimal pH and temperature of the dextrin-uricase conjugate was 9.0 and 45 °C, respectively, whereas the optimal pH and temperature of free uricase was 8.5 and 45 °C, respectively. Furthermore, dextrin-uricase conjugate was more resistant to simulated physiological conditions and trypsin. These results suggested that the stability of uricase could be ameliorated by conjugation with dextrin.
研究领域:
生物化学
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医学
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高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2021
·Vol. 413
·期 26
·6571-6580
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IF 4.157
Liu Donghao,Yang Ping,Wang Fangqi,Wang Cong,Chen Li,Ye Sijing,Dramou Pierre,Chen
Jianhua,He Hua
Nanozymes were the novel research field to replace natural enzymes because of stability and low cost. However, the research on nanozymes was mainly focused on peroxidase, and there was little research about nanozymes with oxidase-like activity, especially mimic oxidase of small molecules related to human physiology. High levels of uric acid (UA) in the body can cause hyperuricemia and gout. And natural uricase cured this disease because it could oxidize UA. The oxidase-like activity of mixed valence state metal organic frameworks with cerium (MVSM) had been studied, but MVSM was found to have uricase-like activity in this article. The catalytic process of UA with MVSM was studied by a variety of analytical methods, which was similar to the natural uricase except for further oxidation of H 2 O 2 . The catalytic activity constants of MVSM were acquired by the Michaelis-Menten equation. MVSM had a better ability to catalyze UA in in vivo and in vitro experiments. An enzyme-free analysis–based mimic uricase for UA was established. All the experimental results proved that MVSM had a good prospect to replace the natural uricase. Graphical abstract
A nanomaterial, mixed valence state Ce-MOF (MVSM), with uricase-like activity has been found in vivo and in vitro. This material has potential to be a fluorescent analysis for detecting uric acid without uricase.
研究领域:
生物化学
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医学诊断
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材料科学
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高影响因子
Nature Communications
·
2025
·Vol. 16
·期 1
·1-15
·
IF 15.7
Liu Lu,Li Xiurong,Chen Yongxian,Gao Junbin,Jiang Yuejun,Ye Yicheng,Wang Peng,Peng Fei,Tu Yingfeng
Uricase (UOx)-based gout treatments are generally limited due in part to the accumulation of H 2 O 2 in an arthrosis environment characterized by a sluggish metabolism. Here we develop a self-cascade nanomotor with high efficiency toward simultaneous uric acid (UA) degradation and H 2 O 2 elimination on the basis of UOx and sodium citrate loading in amine functionalized hollow mesoporous silica nanoparticles (AHMSNs). Due to the inherent asymmetry of AHMSNs, the developed nanomotors can be actuated by ionic diffusiophoresis induced by the enzymatic UA degradation, thus enlarging the diffusion range within the joint cavity. H 2 O 2 generated during UA decomposition can be simultaneously eliminated by the loaded sodium citrate, resulting in a considerable improvement in mammalian cell viability. In vivo studies display powerful therapeutic effect in gouty models using this self-cascade nanomotor system. These results present the potential of nanomotors as active therapeutic agents for gout, providing a perspective on the application of enzyme-driven micro/nanomotors.
研究领域:
医学
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纳米技术
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生物工程
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高影响因子
Journal of Materials Chemistry B
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2013
·Vol. 1
·期 19
·2482-2488
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IF 0
Weifeng Lin,Hui Zhang,Jiang Wu,Zhen Wang,Haotian Sun,Jiang Yuan,Shengfu Chen
PEGylation of protein drugs has thus far been the most important method used in improving the stability and the circulation time, while lowering the immunogenicity of protein drugs. However, the loss of bioactivity of PEGylated protein drugs and the low recovery through the complicated separation process are the most challenging issues. To overcome these problems, a zwitterionic block copolymer poly(methyl acrylic acid-b-sulfobetaine methacrylate) (PMAA-b-PSBMA or abbreviated to PMS) has been chosen to modify a protein drug (uricase) since zwitterionic materials are superior to polyethylene glycol (PEG) in resistance to nonspecific protein adsorption, chemical stability and also low cytotoxicity. As indicated by the results, the conjugation between PMS and uricase could be achieved under 0.03 mg mL−1 of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) and 0.018 mg mL−1N-hydroxysuccinimide (NHS) through weak charge-induced adsorption of PMS on uricase, which is about 0.3% of the concentration for conventional protein–polymer conjugation. The conjugates, efficiently separated from the reaction solution and re-suspended in physiological solution through pH adjustment from pH 5.35 to 7.4, show about 133% of the original activity and better anti-trypsin digestion than the native uricase. These results demonstrate that the conjugation of a zwitterionic copolymer with a short poly(methyl acrylic acid) block could improve the stability of an enzyme without destructively affecting its bioactivity. This may open a new way for the delivery of therapeutic protein drugs.
研究领域:
生物化学
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药物化学
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材料科学
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PubMed
BIOSENSORS & BIOELECTRONICS
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2021
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IF 10.618
Xiuguang Xing,Bangben Yao,Qian Wu,Ru Zhang,Li Yao,Jianguo Xu,Guangheng Gao,Wei Chen
A convenient, fast and non-invasive portable electrochemical uricometer (PUM) assisted with the uricase-packaged nanoflowers (NFs) was constructed for continually and accurately monitoring of uric acid (UA) in urine samples at random intervals in just 20 s. Only a small amount of urine (50 μL) was needed for each test. Electrochemical deposition was adopted to modify gold nanoparticles (AuNPs) on screen-printed carbon electrodes (SPCE) and uricase-inorganic hybrid NFs (UOx-NFs) induced by calcium ions (Ca 2+ ) were introduced for UA detection with expected specificity. Cyclic voltammetry (CV) (detection limit of 8.87 μM and liner range of 0–4 mM) and amperometry (detection limit of 0.82 μM and liner range of 0–5 mM) protocols were studied for UA detection, respectively. Finally, the uric acid in urine had be successfully continually monitored from volunteers with various dietary choosing, the results of which can be adopted as the effective evidence for uric acid control.
研究领域:
医学
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生物传感器
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电化学分析
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高影响因子
Nanoscale
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2017
·Vol. 9
·期 44
·17561-17570
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IF 7.367
Xiao Liu,Wei Qi,Yuefei Wang,Rongxin Su,Zhimin He
Metal–organic frameworks (MOFs) have drawn extensive research interest as candidates for enzyme immobilization owing to their tunable porosity, high surface area, and excellent chemical/thermal stability. Herein, we report a facile and universal strategy for enzyme immobilization using highly stable hierarchically porous metal–organic frameworks (HP-MOFs). The HP-MOFs were stable over a wide pH range (pH = 2–11 for HP-DUT-5) and met the catalysis conditions of most enzymes. The as-prepared hierarchical micro/mesoporous MOFs with mesoporous defects showed a superior adsorption capacity towards enzymes. The maximum adsorption capacity of HP-DUT-5 for glucose oxidase (GOx) and uricase was 208 mg g−1 and 225 mg g−1, respectively. Furthermore, we constructed two multi-enzyme biosensors for glucose and uric acid (UA) by immobilizing GOx and uricase with horseradish peroxidase (HRP) on HP-DUT-5, respectively. These sensors were efficiently applied in the colorimetric detection of glucose and UA and showed good sensitivity, selectivity, and recyclability.
研究领域:
生物化学
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材料科学
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高影响因子
Small
·
2025
·
IF 12.1
Mingchen Lv,Jiaxi Xu,Ran Chen,Wei Hu,Yuxiao Zhou,Min Sun,Zhen Fan,Jianzhong Du
Hyperuricemia, characterized by elevated uric acid levels, is the primary cause of gout. Recombinant uricase is one of the last-resort therapies but generates unwanted pro-inflammatory H 2 O 2 and anti-uricase antibodies. In this work, we developed an albumin corona-coated enzyme-loaded zeolitic imidazolate framework (UCZIF) to sustainably maintain low blood uric acid level without producing H 2 O 2 . The corona coating not only preserves loaded enzymes but also reduces macrophage phagocytosis by 73.4% compared to free uricase. In addition, the uptake level of UCZIF by dendritic cells is reduced by 74.1%, and the maturation of dendritic cells is inhibited by 35.4% compared to free uricase. Animal experiments demonstrate that albumin corona-coated UCZIF effectively lowers blood uric acid level in both acute and diet-induced chronic hyperuricemia models with significantly increasing the half-life of uricase. Furthermore, compared to the generation of anti-uricase antibodies during standalone uricase treatment, the levels of anti-uricase immunoglobulins are significantly reduced by 65.5% (immunoglobulin M) and 76.3% (immunoglobulin G) with repeated administration of albumin corona-coated UCZIF. Overall, this albumin corona-coated nanoscale metal–organic framework offers a promising approach to minimize the immunogenicity induced by exogenous enzymes and further safely reduce uric acid levels in the treatment of hyperuricemia.
研究领域:
生物医学工程
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药物递送
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免疫学
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高影响因子
PROTEIN JOURNAL
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2016
·Vol. 35
·期 4
·318-329
·
IF 1.029
Wu Jing,Yang Xiaolan,Wang Deqiang,Hu Xiaolei,Liao Juan,Rao JingJing,Pu Jun,Zhan Chang-guo,Liao Fei
Prior to the exponential decrease of activity of a uricase from Candida
sp. during storage at 37 °C, there was a plateau period of about 4 days at pH 7.4, 12 days at pH 9.2, and about 22 days in the presence of 30 μM oxonate at pH 7.4 or 9.2, but no degradation of polypeptides and no activity of resolved homodimers. To reveal determinants of the plateau period, a dissociation model involving a serial of conformation intermediates of homotetramer were proposed for kinetic analysis of the thermoinactivation process. In the dissociation model, the roles of interior noncovalent interactions essential for homotetramer integrity were reflected by an equivalent number of the artificial weakest noncovalent interaction; to avoid covariance among parameters, the rate constant for disrupting the artificial weakest noncovalent interaction was fixed at the minimum for physical significance of other parameters; among thermoinactivation curves simulated by numerical integration with different sets of parameters, the one for least-squares fitting to an experimental one gave the solution. Results found that the equivalent number of the artificial weakest noncovalent interaction primarily determined the plateau period; kinetics rather than thermodynamics for homotetramer dissociation determined the thermoinactivation process. These findings facilitated designing thermostable uricase mutants.
研究领域:
生物化学
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酶工程
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2012
·
IF 3.898
Yi Huang,Lijuan Bu,Wen Wang,Xiaoli Qin,Zou Li,Zhao Huang,Yingchun Fu,Xiaoli Su,Qingji Xie,Shouzhuo Yao
Chemical oxidation of glycoprotein-bound thiophene-3-boronic acid (TBA) monomer by relatively mild oxidant chloroplatinate in neutral aqueous solution containing uricase yielded uricase–PTBA–Pt nano bionanocomposite with high enzymatic specific activity on a platinized (Pt plate ) Au electrode, which was then covered by an outer-layer chitosan (CS) film to fabricate a CS/uricase–PTBA–Pt nano /Pt plate /Au electrode. This electrode exhibited a linear amperometric response to uric acid concentration from 5 μM to 1.2 mM with a sensitivity of 134 μA mM −1 cm −2 , a limit of detection ( S / N = 3) of 1 μM, and excellent operation/storage stability, which also worked well in serum samples.
研究领域:
生物化学
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分析化学
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材料科学
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高影响因子
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
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2022
·
IF 3.141
YuLiang Jiao,YuYing Zhu,ShuMin Zeng,ShuFang Wang,Jing Chen,XiangHong Zhou,GuiZhen Ma
A novel uricase producing marine bacterium Priestia flexa alkaAU was isolated and identified. The 16S rDNA and the uricase coding gene were sequenced, analyzed and submitted to GenBank. The uricase from Priestia flexa alkaAU (PFU) was purified, determined to be 58.87 kDa, and conjugated with carboxymethyl chitosan (CMCS) by ionic gelation. CMCS conjugation had no effect on the optimum pH of PFU but decreased the optimum temperature by 10 °C. CMCS conjugation increased the specific activity of PFU by 53% at the human body temperature (37 °C) and small intestine’s pH (pH 6.8). Uricase thermostabilizing ability of CMCS was significant in the range of 37–80 °C but not at lower temperatures. For improvement of the pH stability of PFU, CMCS was more effective at pHs 3–5 than pHs 6–11. CMCS increased the half-life of PFU against artificial intestinal fluid by 1.5 folds, which demonstrated the potential capability of CMCS–PFU for oral administration.
研究领域:
海洋微生物学
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酶学
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生物技术
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药学
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PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Qi Sun,Shiji Miao,Wenlong Yu,En-Yu Jiang,Mixue Gong,Genyan Liu,Xiaogang Luo,Ming-Zhi Zhang
Uric acid (UA) serves as an important biochemical marker of various diseases, making the development of a novel method for its rapid and straightforward visual detection highly valuable. In this study, a uricase-based cellulose membrane biosensor ( UCMB ) was constructed by immobilizing uricase via a Schiff base reaction and nitroblue tetrazolium chloride (NBT) through adsorption. The UCMB detects UA through a mechanism in which uricase catalyzes the oxidation of UA, generation O 2 −· radicals that subsequently oxidize NBT to formazan, producing a distinctive color change from yellow to purple. The UCMB demonstrated successful visual detection of UA within 15 min, allowing for rapid naked-eye analysis. Additionally, the biosensor quantitatively detected UA over a broad linear range from 0 to 1000 μM, with a low detection limit of 3.88 μM. Most notably, the UCMB has accurately measured UA in human serum samples, comparable to the results from a commercial UA meter. These findings suggest that the UCMB can serve as a simple and reliable tool for early diagnosis of UA-related diseases.
研究领域:
分析化学
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生物化学
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生物医学工程
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高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
·
IF 4.4
Yu Xiao,Na Huang,Jiahui Wen,Dan Yang,Huanhuan Chen,Yijuan Long,Huzhi Zheng
Fluorescent bovine serum albumin-protected gold nanoclusters ( [email protected] NCs) can catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to produce blue oxTMB for its peroxidase-like activity. The two absorption peaks of oxTMB overlapped with the excitation and emission peaks of [email protected] NCs, respectively, causing efficient quenching on the fluorescence of [email protected] NCs. The quenching mechanism can be attributed to the dual inner filter effect (IFE). Based on the dual IFE, [email protected] NCs were utilized as both peroxidase mimics and fluorescent reporters for H 2 O 2 detection and further for uric acid detection with uricase. Under optimal detection conditions, the method can be used to detect H 2 O 2 ranging 0.50–50 μM with a detection limit of 0.44 μM and UA ranging 0.50–50 μM with a detection limit of 0.39 μM. The established method had been successfully utilized for the determination of UA in human urine, with massive potential in biomedical applications.
研究领域:
分析化学
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纳米技术
·
生物医学工程
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PubMed
高影响因子
危险品化学品经营许可证（带存储）