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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DOI
高影响因子
Nanomaterials
·
2021
·Vol. 11
·期 9
·2171
·
IF 5.076
Le Wang,Pengxue Sun,Yiyu Yang,Hanzhen Qiao,Hailong Tian,Dapeng Wu,Shuoye Yang,Qipeng Yuan,Jinshui Wang
The field of enzyme cascades in limited microscale or nanoscale environments has undergone a quick growth and attracted increasing interests in the field of rapid development of systems chemistry. In this study, alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and mesoporous silica nanoparticles (MSN) immobilized nicotinamide adenine dinucleotide (NAD+) were successfully immobilized on the zeolitic imidazolate frameworks (ZIFs). This immobilized product was named ZIF@ADH/NAD-MSN/LDH, and the effect of the multi-enzyme cascade was studied by measuring the catalytic synthesis of lactic acid. The loading efficiency of the enzyme in the in-situ co-immobilization method reached 92.65%. The synthesis rate of lactic acid was increased to 70.10%, which was about 2.82 times that of the free enzyme under the optimal conditions (40 °C, pH = 8). Additionally, ZIF@ADH/NAD-MSN/LDH had experimental stability (71.67% relative activity after four experiments) and storage stability (93.45% relative activity after three weeks of storage at 4 °C; 76.89% relative activity after incubation in acetonitrile-aqueous solution for 1 h; 27.42% relative activity after incubation in 15% N, N-Dimethylformamide (DMF) solution for 1 h). In summary, in this paper, the cyclic regeneration of coenzymes was achieved, and the reaction efficiency of the multi-enzyme biocatalytic cascade was improved due to the reduction of substrate diffusion.Keywords:confinement effect;multi-enzyme catalytic cascade;in-situ synthesis;coenzyme recycling;co-immobilization enzyme
研究领域:
生物化学
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纳米生物学
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系统化学
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DOI
PubMed
高影响因子
Nanomaterials
·
2018
·Vol. 8
·期 2
·127
·
IF 3.504
Sen Lin,Shiyong Sun,Ke Wang,Kexuan Shen,Biaobiao Ma,Yuquan Ren,Xiaoyu Fan
The bioinspired design and construction of enzyme@capsule microreactors with specific cell-like functionality has generated tremendous interest in recent years. Inspired by their fascinating complexity, scientists have endeavored to understand the essential aspects of a natural cell and create biomimicking microreactors so as to immobilize enzymes within the hierarchical structure of a microcapsule. In this study, simultaneous encapsulation of alcohol dehydrogenase (ADH) was achieved during the preparation of microcapsules by the Pickering emulsion method using amphiphilic modified TiO2nanoparticles (NPs) as building blocks for assembling the photocatalytic microcapsule membrane. The ADH@TiO2NP microreactors exhibited dual catalytic functions, i.e., spatially confined enzymatic catalysis and the membrane-associated photocatalytic oxidation under visible light. The sustainable cycling of nicotinamide adenine dinucleotide (NAD) coenzyme between NADH and NAD+was realized by enzymatic regeneration of NADH from NAD+reduction, and was provided in a form that enabled further photocatalytic oxidation to NAD+under visible light. This bioinspired ADH@TiO2NP microreactor allowed the linking of a semiconductor mineral-based inorganic photosystem to enzymatic reactions. This is a first step toward the realization of sustainable biological cycling of NAD+/NADH coenzyme in synthetic functional microsystems operating under visible light irradiation.Keywords:enzyme encapsulation;bioinspired microreactors;nicotinamide coenzymes cycling;alcohol dehydrogenase;TiO2nanoparticles
研究领域:
生物化学
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材料科学
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光催化
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DOI
PubMed
高影响因子
BIOTECHNOLOGY AND BIOENGINEERING
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2024
·
IF 3.5
Han-Yu Liu,Pan Ning,Feng Qian,Yao-Wu Wang,Hai-Min Zhang,Pu Wang
Flavin-dependent halogenase (FDH) is highly prized in pharmaceutical and chemical industries for its exceptional capacity to produce halogenated aromatic compounds with precise regioselectivity. This study has devised a multi-enzyme self-assembly strategy to construct an effective and reliable in vitro coenzyme cycling system tailored for FDHs. Initially, tri-enzyme self-assembling nanoclusters (TESNCs) were developed, comprising glucose dehydrogenase (GDH), flavin reductase (FR) and FDH. The TESNCs exhibited enhanced thermal stability and conversion efficiency compared to free triple enzyme mixtures during the conversion of L -Trp to 6-Cl- L -Trp, resulting in a 2.1-fold increase in yield. Subsequently, an ordered co-immobilization of GDH, FR, and FDH was established, further amplifying the stability and catalytic efficiency of the FDH coenzyme cycle system. Compared to the free TESNCs, the immobilized TESNCs demonstrated a 4.2-fold increase in catalytic efficiency in a 5 mL reaction system. This research provides an effective strategy for developing a robust and efficient coenzyme recycling system for FDHs.
研究领域:
生物化学
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合成生物学
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酶工程
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DOI
PubMed
高影响因子
Nanoscale
·
2019
·Vol. 11
·期 36
·16860-16867
·
IF 6.97
Jing-Wen Gao,Miao-Miao Chen,Wei Wen,Xiuhua Zhang,Shengfu Wang,Wei-Hua Huang
Electrochemiluminescence (ECL) nanomaterials are usually deposited compactly on the surface of electrodes, which may cause poor mass transfer of reactants, thereby resulting in low ECL efficiency. In this work, we developed a novel kind of luminescent material denoted as C–Au-luminol nanospheres (C–Au-Lum NSs) by high dispersion of luminophores on porous carbon nanospheres (PCNSs). C–Au-Lum NSs were facilely prepared by the in situ reduction of chloroauric acid with the luminescent reagent luminol (Lum) on the nano-pores of PCNSs. Plenty of luminescent Au-Lum NPs were dispersedly concentrated inside the numerous pores and hollow interiors of PCNSs, effectively increasing the mass transfer of reagents and accelerating the electron transport inside the porous nanospheres. This greatly improved the availability of luminophores and endowed C–Au-Lum NSs with excellent ECL emission. After further integrating with enzymatic circulation and strand displacement, an ultrasensitive ECL biosensor was achieved for the ultrasensitive detection of an important tumor biomarker, mucin1. The logarithmically linear range from 0.1 pg mL−1 to 1 ng mL−1 with the detection limit of 47.6 fg mL−1 (S/N = 3) was achieved, demonstrating the superior performance of C–Au-Lum NSs. This work would provide new ideas for the construction of high-performance ECL sensing platforms for diverse applications.
研究领域:
纳米技术
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生物传感器
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分析化学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
Catalysts
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2025
·Vol. 15
·期 12
·
IF 4
Yang Yang,Kangkang Shang,Xiaorui Gao,Xingmiao Zhu,Mengying Ling,Pu Zheng,Shichao Xu,Pengcheng Chen
Raspberry ketone (RK) is the primary aromatic compound in raspberry fruit, which is widely utilized in perfume, cosmetics, and food additive industries. Currently, RK is predominantly produced chemically. RK biosynthesis through enzyme or whole cell has garnered significant attention due to the mild reaction conditions and the process being regarded as ‘natural’. This study proposed a ‘dual-microorganism, two-phase’ stepwise cascade strategy to produce RK from an economical precursor, 4-hydroxybenzaldehyde (4-HBD). An acetone-tolerant deoxyribose-phosphate aldolase DERAEc(S238D) mutant was obtained through a site-specific rigidification strategy for converting 4-HBD to 4-hydroxybenzylaceton (4-HBA). Then, an engineeredE. colico-expressing isocitrate dehydrogenase and raspberry ketone synthaseRiRZS1 with a citrate-sodium citrate buffer to recycle nicotinamide adenine dinucleotide phosphate (NADPH) was constructed for the conversion of 4-HBA to RK. The final concentration of RK was 50.00 ± 1.92 mmol·L−1with a yield of 86.96%. This strategy provides a scalable coenzyme self-recycling and two-phase catalysis platform for high-value phenolic compounds.
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DOI
高影响因子
ANALYST
·
2023
·
IF 4.2
Li He,Ciping Chen,Yongge Liu,Hong Hai,Jianping Li
A novel electrochemiluminescence(ECL) aptamer biosensor with high sensitivity and selectivity for the detection of tumor biomarker carbohydrate antigen CA125 was constructed, and a strategy of triple amplification of signals was proposed by using exonuclease cyclic cleavage aptamer, combined with rolling ring amplification technologies, generating multi-branched dendritic double-stranded DNA to load a large number of probes through "strand self-growth". The double-stranded DNA, which abbreviated as CP/CA dsDNA, formed by hybridizing the single strand of capture DNA (CP DNA) with the single strand DNA of the CA125 aptamer(CA Apt) was modified on the Fe3O4@Au. When the carbohydrate antigen CA125 was added, CP/CA dsDNA were unwound, and CA125 specifically combined with CA Apt to form a protein-aptamer complex, leaving only CP DNA on the surface of the Fe3O4@Au. RecJf exonuclease cleaved the aptamer in the protein–aptamer complex and released CA125, which recombined to other CA125 aptamers, to form a cycle that produces more CP DNA on the Fe3O4@Au. Three ssDNA (H1, H2, H3) were introduced and hybridized with CP DNA to form a dsDNA with "+" configuration structure. Then phi29 DNA polymerase, T4 DNA ligase, deoxy-ribonucleoside triphosphate (dNTPs) and padlock probes were added to form a large number of complementary strands of padlock probes (CS padlock probes) by rolling cyclic amplification. CS padlock probes were linked to the "+" type dsDNA; then the ssDNA H4 was added and hybridized with CS padlock probe to form multi-branched dendritic dsDNA. A large number of Tris(2,2'-bipyridyl)ruthenium(II) probes were embedded in the double strands, resulting in an extremely strong ECL signal in the presence of the co-reactant tri-n-propylamine (TPA). There is a linear relationship between the ECL signals and CA125 concentrations in the range of 1.0×10-15~1.0×10-8 mg/mL, and the detection limit was 2.38 × 10-16 mg/mL. It has been used for the determination of CA125 in serum samples.
研究领域:
生物传感器
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分析化学
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癌症标志物
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Bioresources and Bioprocessing
·
2022
·Vol. 9
·期 1
·1-9
·
IF 4.983
Ma Yue,Li Tianzhen,Tan Zijian,Ma Long,Liu Haifeng,Zhu Leilei
Catalytic valorization of raw glycerol derived from biodiesel into high-value chemicals has attracted great attention. Here, we report chemoenzymatic cascade reactions that convert glycerol to lactic acid and glycolic acid. In the enzymatic step, a coenzyme recycling system was developed to convert glycerol into 1,3-dihydroxyacetone (DHA) with a yield of 92.3% in potassium phosphate buffer (300 mM, pH 7.1) containing 100 mM glycerol, 2 mM NAD + , 242 U/mL glycerol dehydrogenase-GldA and NADH oxidase-SpNox K184R at 30 °C. Subsequently, NaOH or NaClO 2 catalyzes the formation of lactic acid and glycolic acid from DHA. The high yield of lactic acid (72.3%) and glycolic acid (78.2%) verify the benefit of the chemoenzymatic approaches. Graphical Abstract
研究领域:
化学工程
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生物技术
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催化
·
生化
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DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Xiuhua Zhang,Heqin Liu,Wanrong Hong,Yumin Wang,Jin-Wen Liu,Yingfeng Qin
In this work, a novel fluorescence nanoprobe based on MnO 2 nanosheets and DNAzyme cyclic amplification was developed for label-free detection of GSH. In our design, the DNA@MnO 2 nanoprobe was facilely prepared through the adsorption of DNA probes on the surface of MnO 2 nanosheets. Importantly, MnO 2 nanosheets in the DNA@MnO 2 nanoprobe can act as the recognizer of analytical target GSH. In the presence of GSH, the DNA@MnO 2 nanoprobe are decomposed owing to redox-responsive reaction between GSH and MnO 2 nanosheets, resulting in the generation of abundant Mn 2+ and the desorption of DNA probes. Subsequently, the Mn 2+ -dependent DNAzyme recycling amplification was rapidly initiated and a large number of the blocked G-rich sequences in DNA probes was released. The G-rich sequences were efficiently folded into G-quadruplex structures, which can remarkably light up the fluorescence intensity of thioflavin T (ThT). Experimental data demonstrated that the DNA@MnO 2 nanoprobes for GSH detection have a wide linear range from 0 to 2000 µM. Moreover, this fluorescence sensing strategy was successfully applied for the detection of GSH in complex biological media with satisfying results, providing a simple, facile and cost-efficient nanoplatform for GSH-related clinical disease diagnosis.
研究领域:
分析化学
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纳米技术
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生物医学工程
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生物化学
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产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Fan Xia,Jian Yang,Jingwen Chen,Ximeng Liu,Zhefan Ma,Jinlou Gu
Although hierarchically porous zeolitic imidazolate frameworks (HPZIFs) not only inherit the intrinsic architectural and chemical stabilities of their microporous counterparts but also afford open space for the efficient mass diffusion of the macromolecules involved, their rational design and construction are still challenging. Herein, HPZIFs with tailorable pore sizes ranging from 18 to 54 nm were successfully fabricated by using a newly developed soft-template-directed strategy. Our success rooted in the fact that the screened PS81–PVP44–PEO113 triblock copolymer could effectively coordinate with the metal precursor for the directed crystallization of ZIFs along surfactant assemblies. The advantages of continuous large pores and open structures in such HPZIFs were fully taken into account to serve as a bioreactor for the efficient immunoassay. The expanded large pores provided not only a significantly vast surface area to enhance the density of capture antibodies but also enough space for accommodating multiple conjugated biomolecules in one pore channel. In combination with a cascade enzyme cycle amplification strategy, a model biomarker of prostate-specific antigen (PSA) at the femtomolar level was checked with a limit of detection of 92 fM using the developed immunosensor. Specific screening on patients with prostate cancer or even benign prostatic hyperplasia was exemplified through accurately quantifying small changes of PSA concentration in clinical serum samples, prefiguring the great potential of the developed HPZIF-8 immunosensor platform for the early monitoring and diagnostics of diseases.
研究领域:
材料科学
·
分析化学
·
生物传感器
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生物医学工程
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Yingshu Guo,Yujie Wang,Guangxu Yang,Jing-Juan Xu,Hong-Yuan Chen
Here, a mass-sensitive microRNA sensing surface is developed by utilizing a probe DNA/gold nanoparticles (GNP)/dendrimer composite coupling with an enzymatic amplification process. The probe DNA/GNP/dendrimer composite is prepared via the covalent coupling between the NH 2 groups in PAMAM or DNA and the COOH group on GNP. Target microRNA binds to a stem-loop-structured DNA on maganatic NPs, forming a heteroduplex . By enzyme recycling amplification, a large number of linker sequences are produced on MNPs . Via the combination of probe DNA , the linker DNA on MNPs and a capture DNA on gold chip, the DNA/GNP/dendrimer probe could be assembled on the surface of gold chip inducing a sensitive frequency response. It presents an excellent performance in microRNA-203 quantification with a detection linear range of 1.0×10 −11 –1.0×10 −9 M. Both the success in discriminating expressing miroRNA-203 in MCF-7 cell and the well agreement with the commercial qRT-PCR detection method implied its potential application in early cancer diagnosis for future.
研究领域:
生物传感器
·
纳米技术
·
分子生物学
·
癌症诊断
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）