MICROCHIMICA ACTA
·
2017
·Vol. 184
·期 8
·2941-2948
·
IF 4.58
Han Cong,Li Rongguo,Li Hui,Liu Su,Xu Chenggong,Wang Jingfeng,Wang Yu,Huang Jiadong
The authors describe a label-free electrochemical aptasensor for ultrasensitive and highly specific detection of the antibiotic based on cascade enzymatic recycling couple with DNAzyme amplification. The assay involves two sequential reactions: the first reaction is a λ exonuclease-assisted cyclic digestion reaction triggered by target-aptamer binding. The second reaction is a nicking endonuclease-aided cyclic nicking reaction, which produces a large amount of G-rich nucleic acid segments. These form a G-quadruplex/hemin complex in the presence of K(I) ions and hemin. Because the G-quadruplex/hemin complex acts as a horseradish peroxidase-mimicking DNAzyme with excellent redox activity, the electrochemical signal transduction is accomplished due to the electroreduction of H 2 O 2 . It appears that this work is the first example that cascade enzymatic recycling coupled to DNAzyme amplification is used for antibiotic detection. The aptasensor was applied to the quantitation of kanamycin and gave a response that is linear in the 1 pM to 10 nM kanamycin concentration range, with a detection limit as low as 0.5 pM. The working voltage (vs. Ag/AgCl) at which data can be acquired best is −0.35 V. The assay offers the advantages of remarkably improved sensitivity, use of affordable instrumentation, and simplified operation without the need for electrochemical labeling or addition of labile reagents. Thus, cascade enzymatic recycling coupled to DNAzyme amplification represents a versatile platform for highly sensitive and specific antibiotics detection.
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研究领域:
分析化学
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生物医学工程
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分子生物学
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高影响因子
Energies
·
2022
·Vol. 15
·期 12
·4242
·
IF 3.252
Xue-Yong Li,Jia-Ying Huang,Qiang Zhou,Yuan-Yuan Xu,Ponnandy Prabhu,Ye-Wang Zhang
Acetoin, a four-carbon hydroxyl-keto compound, is used in the food, pharmaceutical, and chemical industries. The cascade enzymatic production is considered a promising and efficient method to produce acetoin. However, the stability and compatibility of the enzymes under the same catalytic conditions are challenges that need to be resolved. In this work, alcohol dehydrogenase, acetaldehyde lyase, and NADH oxidase were selected to work at the same conditions to efficiently convert ethanol into acetoin. These three enzymes were immobilized on epoxy-modified magnetic nanomaterials to obtain highly stable biocatalysts. The stability and the immobilization conditions, including temperature, pH, enzyme–carrier ratio, and immobilization time, were optimized to obtain the immobilized enzymes with a high catalytic activity. The cascade reactions catalyzed by the immobilized enzymes yielded a high conversion of 90%, suggesting that the use of immobilized enzymes is a promising way to produce acetoin.Keywords:immobilization;NADH oxidase;acetaldehyde lyase;alcohol dehydrogenase;acetoin;cascade enzymatic reactions
研究领域:
生物技术
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酶学
·
化学工程
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DOI
高影响因子
TALANTA
·
2025
·
IF 6.1
Kai Liu,Yuru Wang,Haibing Zhu,Feng Shi,Zijun Lai,Yan Long,Chuanli Ren,Juan Li,Zhanjun Yang
The metal-organic frameworks (MOFs)-derived nanozymes in air atmosphere have gained great attention in biosensing fields. Nevertheless, this derivative pattern may result in the destabilization of the MOF framework and the aggregation of active sites, consequently diminishing its catalytic activity. Herein, we reported an inert-remodeling strategy to build bimetal-confined nitrogen-doped carbon nanozyme for dual-mode cascade enzyme biosensing. The strategy was easily achieved by pyrolysis of MOFs (CoNi-ZIF-67 as model) precursor in argon atmosphere, leading to the formation of CoNi bimetallic nanoparticles uniformly confined nitrogen-doped carbon (CoNi–CN) nanozyme. This derivative nanozyme exhibits significantly enhanced peroxidase (POD)-like activity, which is 4 times higher than that of NiCo 2 O 4 nanozyme (CoNi-ZIF-67 derivative in air atmosphere) and 54 times higher than that of CoNi-ZIF-67 precursor. The excellent POD-like activity of CoNi–CN nanozyme is ascribed to the following facts: i) integrate structure with uniformly dispersed CoNi bimetal active sites; ii) confinement effect of CoNi bimetal encapsulated in CN architecture. Integrating with glucose oxidase (GOx) to prepare cascade enzyme of CoNi–CN@GOx, colorimetric-chemiluminescent imaging sensor based on CoNi–CN@GOx cascade system was developed for glucose detection. Glucose was assayed in wide linear ranges of 0.08–15 mM (colorimetric) and 0.1–30 mM (CL imaging). This research provides a promising inert-remodeling strategy to construct high-performance nanozyme for dual mode biosensing applications.
研究领域:
生物传感
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材料科学
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分析化学
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DOI
PubMed
高影响因子
ANALYST
·
2019
·Vol. 144
·期 16
·4995-5002
·
IF 4.019
Rufeng Zhang,Yu Wang,Xiaonan Qu,Shasha Li,Yihan Zhao,Fenfen Zhang,Su Liu,Jiadong Huang,Jinghua Yu
Herein, a split G-quadruplex DNAzyme as a signal reporter was integrated into an electrochemical sensing platform for the detection of antibiotics with specificity and sensitivity. To improve the signal-to-noise ratio, two G-rich oligonucleotide sequences (G1 and G2) were blocked into two different hairpin probes, preventing the two segments from assembling into a spilt G-quadruplex structure. Moreover, we designed a double-arch probe, consisting of an aptamer as the recognition element and two-step enzymatic signal amplification. Concretely, the first is the Nt.BbvCI-assisted nicking cyclic reaction activated by target-aptamer binding, and the second is exonuclease III-aided cyclic amplification for generating abundant G1 and G2. The modified capture probe on the electrode was used to combine G1 and G2 to form the spilt G-quadruplex/hemin when K+ and hemin were present. This complex plays the role of DNAzyme with superior horseradish peroxidase activity in catalyzing the decomposition of H2O2. Under optimal conditions, this biosensor showed an excellent performance for sensing kanamycin with a detection limit of 83 fM for kanamycin concentrations ranging from 100 fM to 1 nM. Hence, the proposed strategy has potential as an efficient and actual platform for small molecule analysis.
研究领域:
生物化学
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分析化学
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生物技术
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DOI
PubMed
高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
·
2025
·
IF 9.7
Dan Xia,Tingting Shi,Wuxiu Cao,Baoe Li,Donghui Wang,Chunyong Liang,MingDong Dong
Bacterial infection, hypoxia and oxidative stress caused by high blood glucose are the main problems in diabetic wound healing. The glucose-activated cascade reaction could fundamentally solve these problems in diabetic wounds by consuming glucose and eliminating bacteria. In this paper, glucose oxidase (GOx) was immobilized on polyethylenimine (PEI) adsorbed molybdenum disulfide (MoS 2 ) to prepare MoS 2 -PEI-GOx (MPG) composite particles, which were doped into the self-healing hydrogel of polyvinyl alcohol (PVA) and dextran (Dex) for preparing PD@MPG hydrogel. GOx can decompose excessive glucose into H 2 O 2 and gluconic acid, thereby reducing the pH value and improving the microenvironment of the wound. Low pH value enables MoS 2 exert a similar role as peroxidase, catalyzing H 2 O 2 to produce hydroxyl radicals ( OH) to kill bacteria. After reducing the blood glucose of the wound, MoS 2 can play a similar role as catalase, catalyzing the excess H 2 O 2 at the wound to convert to O 2 , thereby alleviating hypoxia and oxidative stress. In addition, the wound healing ability of PD@MPG has been evaluated by in vivo study. The self-healing hydrogels developed in this study hold promise as an innovative dressing for diabetic wound healing.
研究领域:
生物材料学
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医学
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生物化学
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DOI
PubMed
高影响因子
TALANTA
·
2016
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IF 4.035
Yan Zhao,Ying Wang,Xiaobin Zhang,Rongmei Kong,Lian Xia,Fengli Qu
The ultrasensitive monitoring of glucose with a fast and accurate method is significant in potential therapeutics and optimizes protein biosynthesis . Incorporation of enzyme into matrix is considered as promising candidates for constructing highly sensitive glucose-responsive systems. In this study, three-dimensional poly(acrylic acid) brushes-nanospherical silica (PAA-nano silica) with high amplification capability and stability were used to covalently immobilize bienzymes for cascade enzymatic catalysis. The major advantages of PAA-nano silica-bienzyme co-incorporation is that the enzymes are proximity distribution, and such close confinement both minimized the diffusion of intermediates among the enzymes in the consecutive reaction and improve the utilization efficiency of enzymes, thereby enhancing the overall reaction efficiency and specificity. Thus, this present bienzymatic biosensor shows robust signal amplification and ultrasensitivity of glucose-responsive properties with a detection limit of 0.04 μM.
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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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2022
·
IF 9.221
Shan He,Huiting Lian,Xuegong Cao,Bin Liu,Xiaofeng Wei
Non-invasive body fluid analysis has been increasingly developed for health monitoring and disease diagnostics. However, some portable devices, as one representative non-invasive application, must require complicated multi-step reactions, expensive photoelectric components, and be difficult for large-scale fabrication. Herein, we have integrated a cascaded enzymatic reaction-mediated multicolor pixelated quantitative system on the wearable microfluidic device (McPiQ-μWAD) for non-invasive and sensitive detection of sweat glucose. This is the first application of non-invasive portable detection using multicolor pixelated quantification, and it’s also the first smartphone-assistant multicolor pixelated analysis for Localized Surface Plasmon Resonance (LSPR) of gold nanorods (Au NRs). The wearable device incorporated laser-ablated microfluidic technology and enzyme-mediated Au NRs etching system for the achievement of rapid and sensitive pixelated quantification. The McPiQ-μWAD successfully transferred sweat glucose into a visual colorful signal in the concentration range of 0.02–2.4 mM (0.36–43.2 mg/dL) by pixelated quantification with the limit of detection (LOD) of 0.02 mM (0.36 mg/dL). Compared with commercial glucose kit-based spectrometry, the detection results of real sweat samples from healthy volunteers showed desirable reliability and accuracy based on the McPiQ-μWAD. The presented McPiQ-μWAD provides the high potential to be broadly utilized for non-invasive analyses for applications in family health surveillance and point-of-care tests of emergency.
研究领域:
生物医学工程
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分析化学
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医疗诊断
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DOI
高影响因子
SOLID STATE SCIENCES
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2024
·
IF 3.4
Ying Zhu,Tongkai Ji,Xingwei Cai,Jing Wang
A novel NiO/CeO 2 −mSiO 2 nanocages with superior peroxidase−like activities due to the synergetic effect between NiO and CeO 2 were fabricated by a simple etching−precipitation method with core−shell structured Cu 2 O@mSiO 2 as a sacrificial template. Based on the peroxidase−like activity of the NiO/CeO 2 −mSiO 2 nanocages, an enzyme cascade−triggered colorimetric sensing platform was exploited for alkaline phosphatase (ALP) detection, owing to the fact that ascorbic acid (AA) that is the ALP enzymatic hydrolysis product of ascorbic acid 2−phosphate (AAP) can efficiently reduce the oxidation product of TMB (oxTMB) generated under the catalysis of the NiO/CeO 2 −mSiO 2 . For the sensing of ALP, a good linear response was observed in the range of 0.0005–1.25 mU mL −1 with the limit of detection (LOD) as low as 0.00048 mU mL −1 . Furthermore, the proposed enzyme cascade−triggered colorimetric sensing platform could be further explored for sensing of tyrosinase, according to the fact that dopamine (DA) the catalytic product of tyramine (Tyr) by tyrosinase could initiate the redox reaction with the oxTMB acquired under the catalysis of the NiO/CeO 2 −mSiO 2 . A wide linear response was found for tyrosinase detection from 0.02 mU mL −1 to 3.4 U mL −1 with a LOD as low as 0.002 U mL −1 . This work not only offers a facile and effective approach to fabricate nanozymes with excellent catalytic activity, but also provides a promising enzyme cascade−triggered colorimetric method for the ultra−sensitive detection of biological enzymes.
研究领域:
生物化学
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纳米医学
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分析化学
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DOI
高影响因子
JOURNAL OF MATERIALS SCIENCE
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2023
·Vol. 58
·期 17
·7441-7455
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IF 4.5
Wang Yijie,Li Qiulan,Xia Qinghai,Yang Yaling,Yang Dezhi
Sensitive and effective detection of xanthine oxidase (XOD) is essential for assessing human health. Herein, a dual-enzyme-based cascaded reaction system was developed for xanthine and xanthine oxidase assay based on the peroxidase (POD)-like catalytic activity of Fe 3 O 4 @MWCNTs/Hemin nanocomposite. The XOD could catalyze xanthine to produce H 2 O 2 , which is subsequently converted by Fe 3 O 4 @MWCNTs/Hemin into hydroxyl radicals (·OH) to oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) to produce a light blue. Notably, it is demonstrated that the modification of MWCNTs and Hemin could significantly enhance the POD-like activity from 9.78 × 10 −8 M s −1 ( V max value of Fe 3 O 4 ) to 23.34 10 −8 M s −1 ( V max value of Fe 3 O 4 @MWCNTs/Hemin), which makes the detection of xanthine oxidase more sensitive. By using the smartphone to capture the colorimetric signal of oxidized TMB (oxTMB), the assay can detect xanthine and XOD as low as 0.250 µM and 0.216 mU mL −1 , respectively. The reliability of the method in human serum suggested satisfactory spiked recoveries in the range of 92.350–105%. This work provided a new insight for Fe 3 O 4 -based nanozyme design and monitoring XOD level in blood. Graphical Abstract
研究领域:
生物化学
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纳米技术
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分析化学
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医学诊断
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DOI
高影响因子
Scientific Reports
·
2017
·Vol. 7
·期 1
·1-10
·
IF 4.259
Jia Xiaojing,Liu Ying,Han Yejun
Acetoin (3-hydroxy-2-butanone) is an important bio-based platform chemical with wide applications. In vitro enzyme catalysed synthesis exhibits great feasibility in the production of chemicals with high purity. In the present work, a synthetic pathway involving a two-step continuous reaction was constructed in vitro for acetoin production from pyruvate at improved temperature. Thermostable candidates, acetolactate synthase (coAHASL1 and coAHASL2 from Caldicellulosiruptor owensensis OL) and α-acetolactate decarboxylase (bsALDC from Bacillus subtilis IPE5-4) were cloned, heterologously expressed, and characterized. All the enzymes showed maximum activities at 65–70 °C and pH of 6.5. Enzyme kinetics analysis showed that coAHASL1 had a higher activity but lower affinity against pyruvate than that of coAHASL2. In addition, the activities of coAHASL1 and bsALDC were promoted by Mn 2+ and NADPH. The cascade enzymatic reaction was optimized by using coAHASL1 and bsALDC based on their kinetic properties. Under optimal conditions, a maximum concentration of 3.36 ± 0.26 mM acetoin was produced from 10 mM pyruvate after reaction for 24 h at 65 °C. The productivity of acetoin was 0.14 mM h −1 , and the yield was 67.80% compared with the theoretical value. The results confirmed the feasibility of synthesis of acetoin from pyruvate with a cell-free enzyme catalysed system at improved temperature.
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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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2025
·Vol. 193
·期 1
·31
·
IF 5.3
Zhang Yukai,Yang Mo,Liu Juan
A novel artificial mineralized bioprobe was developed through a templated strategy, enabling highly sensitive and rapid detection of neuron-specific enolase (NSE) in serum samples. This probe was based on a nitrogen-doped carbon porous hollow shell as a carrier, loaded with Fe-Cu nanoparticles exhibiting highly efficient peroxidase-like activity. The Fe-Cu nanoparticles demonstrated outstanding catalytic decomposition of hydrogen peroxide, while the unique porous structure provided an ideal confined space for immobilizing the native enzyme. The acidic microenvironment generated by the enzymatic products of anchored glucose oxidase activates adjacent Fe-Cu bimetallic nanozymes, enabling cascade catalytic reactions under near-neutral conditions. Furthermore, due to the confining effect of the porous carbon shell layer, immobilized natural enzymes showed superior stability in response to temperature changes (10–50 °C) and during storage (30 days). In the presence of NSE targets, immunocomplex formed and catalyzed the cascade decomposition of substrate glucose, which attacked the chromogenic substrate to induce color change. Under optimized conditions, the developed sensor exhibited an ultra-wide linear response range (0.1–100 ng mL − 1 ) and a low detection limit (LOD = 0.0486 ng mL − 1 ). This work provides new insights for developing highly efficient enzyme-catalyzed probes.
Section
Graphical
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PubMed
高影响因子
ACS Sustainable Chemistry & Engineering
·
2025
·
IF 7.3
Yaju Xue,Boxia Guo,Xiuling Ji,Bowen Li,Yuhong Huang
Glycine serves as a crucial element in basic metabolic processes and acts as a precursor in the production of numerous industrial chemicals. Here we developed a streamlined artificial cascade enzyme catalysis pathway for converting CO2 into glycine through improving the CO2 activation and employing direct C–C condensation. After the optimization of enzyme combinations and dose-dependent regulation, the integration of reduced nicotinamide adenine dinucleotide regeneration and ionic liquids were introduced and facilitated the construction of an artificial cascade ionozyme catalysis system. Consequently, the glycine yield reached a remarkable 17.87-fold increase compared to that of the initial reaction system. Additional molecular simulations further demonstrated that the ionozyme possessed enhanced structural stability and substrate affinity, as indicated by an increase in hydrogen bonds and a diminished interference from water molecules in protein–ligand binding. This innovative approach holds great promise for sustainable CO2 bioconversion to glycine and other valuable chemicals.
研究领域:
合成生物学
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生物催化
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二氧化碳利用
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绿色化学
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代谢工程
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DOI
高影响因子
危险品化学品经营许可证（带存储）