ANALYTICAL CHEMISTRY
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2017
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IF 6.32
Juan Zhang,Xiaonan Wang,Tingjun Chen,Chang Feng,Genxi Li
In this work, a new strategy for electrochemical analysis of enzyme has been proposed based on a self-assembled lipid bilayer on an electrode surface mediated by hydrazone chemistry. Taking aldolase as an example, the enzyme can catalyze the formation of products containing carbonyl groups. These groups can react with hydrazine groups of the functional lipid derivative, resulting in the self-assembly of a lipid bilayer on a guanidinium modified electrode surface. The lipid bilayer will then prevent the movement of hydrophilic electrochemical probes. Consequently, the catalytic reaction of the enzyme may result in the change of the obtained electrochemical peak current. Experimental results reveal that aldolase activity can be analyzed over a widely linear detection range from 5 mU/L to 100 U/L with a low detection limit of 1 mU/L. Meanwhile, the method can exhibit good precision and reproducibility and it can be applied for real sample analysis. What is more, because the lipid bilayer is the universal basis for cell-membrane structure, while hydrazone chemistry is popular in nature, this work may also provide a new insight for the development of electrochemical analysis and electrochemical biosensors.
研究领域:
生物化学
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分析化学
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DOI
PubMed
高影响因子
INTERNATIONAL BIODETERIORATION & BIODEGRADATION
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2015
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IF 2.131
Lin-Na Du,Gang Li,Yu-Hua Zhao,He-Kun Xu,Yang Wang,Ying Zhou,Liang Wang
We investigated biodegradation of the azo dye methyl orange (MO) by Aeromonas sp. DH-6. Nearly 100% decolorization of MO (100 mg l −1 ) by the strain was observed at pH 3.0–7.0 and a broad temperature range (5–45 °C) after incubation for 12 h. Most of the tested carbon and nitrogen sources could significantly enhance MO decolorization, but of the metal ions , only Mg 2+ , Mn 2+ , and 1.5 mM of Ca 2+ could increase the decolorization. The strain still showed a high capacity for decolorizing MO even at a concentration of up to 1000 mg l −1 . Based on the gas chromatography-mass spectrometry analysis (GC-MS) and high performance liquid chromatography analysis (HPLC) data, N,N-dimethyl- p -phenylenediamine and 4-aminobenzenesulfonic acid were identified as the main intermediates of MO biodegradation, and a possible degradation pathway was proposed. Moreover, results of enzyme analysis indicated that laccase, NADH-DCIP reductase, and azo reductase were responsible for the degradation. Overall, high biodegradation efficiency at a broad range of temperatures and dye concentrations suggested that strain DH-6 would have high potential in the bioremediation of azo dyes.
研究领域:
环境科学
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微生物学
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染料降解
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
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2022
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IF 8.008
Rui Liu,Gaoyi Yi,Baian Ji,Xiuqiong Liu,Yuanqi Gui,Zhining Xia,Qifeng Fu
Enzyme assays are important for studying enzyme-mediated biochemical reactions and for clinical diagnosis and drug development. The technique of an immobilized enzyme microreactor (IMER) integrated with capillary electrophoresis (CE) has been frequently utilized in online enzyme assays. However, the traditional approaches for IMER-CE enzyme analysis have some defects such as low loading capacity and poor stability. Herein, metal–organic frameworks (MOFs), which have enormous potential in the fields of enzyme immobilization and capillary electrochromatographic (CEC) separation, were first explored as novel support materials with good enzyme immobilization performance and stationary phases with excellent separation abilities to construct an integrated MOFs-IMER-CEC microanalysis system for a high-efficiency online enzyme assay. As a proof-of-concept demonstration, acetylcholinesterase (AChE) was immobilized on a densely packed UiO-66-NH2 nanocrystal coating on a capillary inner surface with abundant intercrystalline mesoporosity and was employed as a highly effective and robust IMER for CEC-integrated online enzyme analysis. The excellent separation performance of the UiO-66-NH2-modified capillary was verified by high-efficiency separation of three types of neutral, acidic, and basic compounds. The Michaelis–Menten constant and enzyme inhibition kinetics of UiO-66-NH2–IMER were systematically assessed, exhibiting distinct advantages such as remarkably increased enzyme loadability, superior affinity for substrates, and greatly improved stability and repeatability compared to CE-integrated IMERs prepared by the traditional covalent bonding method. Furthermore, the developed method was successfully utilized for detecting organophosphorus pesticides in leguminous vegetable samples, demonstrating its strong practicality. The study not only proposed a novel support material and construction strategy for a high-performance microchannel-based IMER but also can be widely used in bioanalysis and biosensing research.
研究领域:
分析化学
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生物化学
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生物技术
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环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2015
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IF 5.636
Lihua Lin,Shengquan Liu,Zhou Nie,Yingzhuang Chen,Chunyang Lei,Zhen Wang,Chao Yin,Huiping Hu,Yan Huang,Shouzhuo Yao
Nowadays, large-scale screening for enzyme discovery, engineering, and drug discovery processes require simple, fast, and sensitive enzyme activity assay platforms with high integration and potential for high-throughput detection. Herein, a novel automatic and integrated micro-enzyme assay (AIμEA) platform was proposed based on a unique microreaction system fabricated by a engineered green fluorescence protein (GFP)-functionalized monolithic capillary column, with thrombin as an example. The recombinant GFP probe was rationally engineered to possess a His-tag and a substrate sequence of thrombin, which enable it to be immobilized on the monolith via metal affinity binding, and to be released after thrombin digestion. Combined with capillary electrophoresis-laser-induced fluorescence (CE-LIF), all the procedures, including thrombin injection, online enzymatic digestion in the microreaction system, and label-free detection of the released GFP, were integrated in a single electrophoretic process. By taking advantage of the ultrahigh loading capacity of the AIμEA platform and the CE automatic programming setup, one microreaction column was sufficient for many times digestion without replacement. The novel microreaction system showed significantly enhanced catalytic efficiency, about 30 fold higher than that of the equivalent bulk reaction. Accordingly, the AIμEA platform was highly sensitive with a limit of detection down to 1 pM of thrombin. Moreover, the AIμEA platform was robust and reliable to detect thrombin in human serum samples and its inhibition by hirudin. Hence, this AIμEA platform exhibits great potential for high-throughput analysis in future biological application, disease diagnostics, and drug screening.
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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医学诊断
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药学
✓ 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）