Biosensors for the evaluation of lipase activity

In the review a comprehensive analysis is given of instrumental analytical approaches to control the activity of lipolytic enzymes and their substrates. A special attention is attached to the methods based on the biosensor technology. A number of electrochemical, optical and mechanical biosensors that were developed in several laboratories are analyzed in detail. In addition, perspectives are evaluated for the development of these biosensors and their practical application. It was concluded that biosensors may provide all practical demands which are in enzymology field at the express determination of lipase activity and triacylglycerol levels in biological objects.     

Electrochemical biosensing based on universal affinity biocomposite platforms

Rigid conducting biocomposites are versatile and effective transducing materials for the construction of a wide range of amperometric biosensors such as immunosensors, genosensors and enzymosensors, particularly if the transducer is bulk-modified with universal affinity biomolecules. The strept(avidin)-graphite-epoxy biocomposite could be considered as an universal immobilization platform whereon biotinylated DNAs, oligonucleotides, enzymes or antibodies can be captured by means of the highly affinity (strept)avidin-biotin reaction. Universal affinity biocomposite-based biosensors offer many potential advantages compared to more traditional electrochemical biosensors commonly based on a biologically surface-modified transducer. The integration of many materials into one matrix is their main advantage. As biological bulk-modified materials, the conducting biocomposites act not only as transducers, but also as reservoir for the biomaterial. After its use, the electrode surface can be renewed by a simple polishing procedure, establishing a clear advantage of these approaches relative to classical biosensors and other common biological assays. Moreover, the same material is useful for the analysis of many molecules whose determinations are based on genetic, enzymatic or immunological reactions. The different strategies for electrochemical genosensing, immunosensing and enzymosensing, all of them being dependent on the presence of a redox enzyme marker for the generation of the electrochemical signal, based on this universal affinity biocomposite platform are all presented and discussed.     

Progress and challenges for biosensors using plant tissue materials

Biocatalytic sensors using plant tissue materials in conjunction with electrochemical elements offer an alternative to biosensors based on isolated enzymes. In favorable cases, such plant based sensors show attractive analytical properties in addition to low cost, simplicity of construction, and reduced co-factor requirements. A review of the current state of the art is provided along with some previously unpublished examples of new biosensors using plant tissue materials.     

Application of ionic liquids in electrochemical sensing systems

Since 1992, when the room temperature ionic liquids (ILs) based on the 1-alkyl-3-methylimidazolium cation were reported to provide an attractive combination of an electrochemical solvent and electrolyte, ILs have been widely used in electrodeposition, electrosynthesis, electrocatalysis, electrochemical capacitor, and lithium batteries. However, it has only been in the last few years that electrochemical biosensors based on carbon ionic liquid electrodes (CILEs) and IL-modified macrodisk electrodes have been reported. However, there are still a lot of challenges in achieving IL-based sensitive, selective, and reproducible biosensors for high speed analysis of biological and environmental compounds of interest. This review discusses the principles of operation of electrochemical biosensors based on CILEs and IL/composite-modified macrodisk electrodes. Subsequently, recent developments and major strategies for enhancing sensing performance are discussed. Key challenges and opportunities of IL-based biosensors to further development and use are considered. Emphasis is given to direct electron-transfer reaction and electrocatalysis of hemeproteins and enzyme-modified composite electrodes.     

微阵列电化学生物传感器研究进展

本文简要介绍了微阵列电化学生物传感器的基本原理和分类,评述了微阵列电化学生物传感器的研究进展。     

Amperometric ATP biosensor based on polymer entrapped enzymes

A dual enzyme electrode for the detection of adenosine-5'-triphosphate (ATP) at physiologically relevant pH levels was developed by co-immobilization of the enzymes glucose oxidase (GOD) and hexokinase (HEX) using pH-shift induced deposition of enzyme containing polymer films. Application of a simple electrochemical procedure for the co-immobilization of the enzymes at electrode surfaces exhibits a major improvement of sensitivity, response time, reproducibility, and ease of fabrication of ATP biosensors. Competition between glucose oxidase and hexokinase for the substrate glucose involving ATP as a co-substrate allows the determination of ATP concentrations. Notable control on the immobilization process enables fabrication of micro biosensors with a diameter of 25 microm. The presented concept provides the technological basis for a new generation of fast responding, sensitive, and robust biosensors for the detection of ATP at physiological pH values with a detection limit of 10 nmol l(-1).     

Bioelectrocatalysis-based application of quinoproteins and quinoprotein-containing bacterial cells in biosensors and biofuel cells

Electrochemical studies on the applied aspects of quinoproteins are briefly reviewed. Catalytic reactions of quinoprotein enzymes can be connected to electrochemical reactions directly or by the mediation of molecules functioning as electron acceptors of the enzymes. Such an enzyme-electrochemical reaction is called bioelectrocatalysis. It provides a novel method of kinetic analysis of enzyme catalysis and even whole bacterial cell catalysis. The principle of bioelectrocatalysis is first described, then, the bioelectrocatalysis-based application of quinoproteins in biosensors is mentioned. Characteristics and performance of this type of biosensor is explained by citing our own work. Possible application in bioreactors and biofuel cells is also mentioned.     

Effect of dimethyl sulfoxide on the structure and the functional properties of horseradish peroxidase as observed by spectroscopy and cyclic voltammetry

Electrochemical biosensors have found wide application in food and clinical areas, as well as in environmental field. A large number of articles focused on horseradish peroxidase (HRP)-based biosensors have been published in the last decade, due to the capability of HRP to quantitatively detect the presence of hydrogen peroxide produced in a reaction. At present a large body of multi-enzymatic amperometric biosensors are realized by entrapping HRP together with other enzymes into a polymeric matrix; these systems represent a promising example of simple, low-cost electrochemical tools for the analysis of bioanalytes in solution, such as glucose, choline and cholesterol. Due to the fact that polymers used for HRP entrapping are soluble in organic solvents and that many solvents are strong denaturants of aquo-soluble proteins, in this paper we investigate (in particular, by circular dichroism and electron paramagnetic spectroscopies) the effect of dimethyl sulfoxide, one of the organic solvents employed for polymer solubilization, on the structure and the functionality of HRP, in order to determine the effect induced by the solvent concentration on the structure and activity of the hemoprotein. This is relevant for basic and applied biochemistry, HRP being largely employed in bioinorganic chemistry and sensor area.     

Electrochemical characteristics of a gold nanoparticle-modified controlled enzyme–electrode contact junction electrode

Biodevices in which biomolecules such as enzymes and antibodies are immobilized on the surface of electrode materials are capable of converting chemical energy into electrical energy, and are expected to contribute to solving energy problems and developing medical measurements especially as biobatteries and biosensors. Device performance depends on the interface formed between the biomolecule layer and electrode material, and the interface is required to simultaneously achieve a highly efficient enzymatic reaction and electron transfer. However, when enzymes were immobilized on a material surface, the enzymes undergoes a structural change due to the interaction between the enzyme and the electrode surface, making it difficult to maximize the function of the enzyme molecule on the material surface. In this study, we postulate that the structural change of the enzyme would be reduced and the electrochemical performance improved by making the contact area between the enzyme and the electrode extremely small and adsorbing it as a point. Therefore, we aimed to develop a high-power biodevice that retains enzyme structure and activity by interposing gold nanoparticles (AuNPs) between the enzyme and the electrode. The enzymatic and electrochemical properties of pyrroloquinoline quinone-dependent glucose dehydrogenase adsorbed on AuNPs of 5–40 nm diameter were investigated. We found that the characteristics differed among the particles, and the enzyme adsorbed on 20 nm AuNPs showed the best electrochemical characteristics.

     

纳米粒子标记DNA 探针在电化学DNA 生物传感器中的应用

介绍了纳米电化学DNA生物传感器的基本概念和分类,并介绍了用于DNA标记的纳米粒子的六种类型及其三大检测方法,在此基础上对纳米电化学DNA生物传感器在基因检测、疾病诊断、DNA检测等方面的最新进展进行了综述与讨论。     

Sensor and biosensor preparation, optimisation and applications of Prussian Blue modified electrodes

Being one of the most commonly used electrochemical mediators for analytical applications, Prussian Blue has found a wide use in the biosensor field during the last years. Its particular characteristic of catalysing hydrogen peroxide reduction has been applied in the construction of a large number of oxidase enzyme-based biosensors for clinical, environmental and food analysis. By modifying an electrode surface with Prussian Blue, it is in fact possible to easily detect hydrogen peroxide at an applied potential around 0.0 V versus Ag/AgCl, thus making possible coupling with oxidase enzymes while also avoiding or reducing electrochemical interferences. Papers dealing with glucose, lactate, cholesterol and galactose biosensors that are based on the use of Prussian Blue have recently appeared in the most important analytical chemistry journals. Another recent trend is the use of a choline probe based on choline oxidase for pesticide determination to exploit the inhibition of acetylcholinesterase by these compounds. In addition, the use of Prussian Blue in the development of biosensors for food analysis has captured the interest of many research groups and led to improved methods for the detection of glutamate, galactose, alcohol, fructosyl amine, formate, lysine and oxalate. This review will focus on the biosensing aspects of Prussian Blue-based sensors giving a general overview of the advantages provided by such mediator as well as its drawbacks. A comprehensive bibliographic reference list is presented together with the most up to date research findings in this field and possible future applications. The commercial potential of sensors based on this mediator will also be discussed.     

Immobilization strategies to develop enzymatic biosensors

Immobilization of enzymes on the transducer surface is a necessary and critical step in the design of biosensors. An overview of the different immobilization techniques reported in the literature is given, dealing with classical adsorption, covalent bonds, entrapment, cross-linking or affinity as well as combination of them and focusing on new original methods as well as the recent introduction of promising nanomaterials such as conducting polymer nanowires, carbon nanotubes or nanoparticles. As indicated in this review, various immobilization methods have been used to develop optical, electrochemical or gravimetric enzymatic biosensors. The choice of the immobilization method is shown to represent an important parameter that affects biosensor performances, mainly in terms of sensitivity, selectivity and stability, by influencing enzyme orientation, loading, mobility, stability, structure and biological activity.     

Biosensing environmental pollution

Whole-cell biosensors are finding increasing use for the detection of environmental pollution and toxicity. These biosensors are constructed through the fusion of promoters, responsive to the relevant environmental conditions, to easily monitored reporter genes. Depending on the choice of reporter gene, expression can be monitored by the production of colour, light, fluorescence or electrochemical reactions. Recent advances in this area have included the development of biosensors of compact size that enable the on-line and in situ monitoring of a large number of environmental parameters.     

Biological and technical challenges for implementation of yeast-based biosensors

Biosensors are low-cost and low-maintenance alternatives to conventional analytical techniques for biomedical, industrial and environmental applications. Biosensors based on whole microorganisms can be genetically engineered to attain high sensitivity and specificity for the detection of selected analytes. While bacteria-based biosensors have been extensively reported, there is a recent interest in yeast-based biosensors, combining the microbial with the eukaryotic advantages, including possession of specific receptors, stability and high robustness. Here, we describe recently reported yeast-based biosensors highlighting their biological and technical features together with their status of development, that is, laboratory or prototype. Notably, most yeast-based biosensors are still in the early developmental stage, with only a few prototypes tested for real applications. Open challenges, including systematic use of advanced molecular and biotechnological tools, bioprospecting, and implementation of yeast-based biosensors in electrochemical setup, are discussed to find possible solutions for overcoming bottlenecks and promote real-world application of yeast-based biosensors.     

Electrochemical reduction of cytochrome P450 as an approach to the construction of biosensors and bioreactors

In the present review an attempt was made to present an up-to-date amount of the data on electrochemical reduction of the hemoprotein cytochrome P450. The concept and potentialities of enzyme electrodes--transducers--as the main element for construction of electrochemical biosensors were discussed. Different types of electrodes for bioelectrochemistry were analysed. New nanotechnological approaches to cytochrome P450 immobilisation were reported. It was shown that nanobiotechnology in electrochemistry has potential application in manufacturing biosensors and bioreactors for clinical medicine and pharmacology.     

Cytochrome P450 biosensors-a review

Cytochrome P450 (CYP) is a large family of enzymes containing heme as the active site. Since their discovery and the elucidation of their structure, they have attracted the interest of scientist for many years, particularly due to their catalytic abilities. Since the late 1970s attempts have concentrated on the construction and development of electrochemical sensors. Although sensors based on mediated electron transfer have also been constructed, the direct electron transfer approach has attracted most of the interest. This has enabled the investigation of the electrochemical properties of the various isoforms of CYP. Furthermore, CYP utilized to construct biosensors for the determination of substrates important in environmental monitoring, pharmaceutical industry and clinical practice.     

Inlaid Nd-substituted bismuth titanate nanoplates for protein immobilization and Nd-controlled electrochemical properties

Neodymium (Nd) substituted bismuth titanate (Bi(4-x)Nd(x)Ti(3)O(12), BNTO-x) nanoplates inlaid one another were prepared by sol-gel hydrothermal method, which was explored for protein immobilization and biosensor fabrication. Comparative experiments witnessed that Bi(3+) ions in bismuth titanate (Bi(4)Ti(3)O(12), BTO) were successfully substituted with Nd(3+) ions, and the electrochemical properties of the Hb-Chi-BNTO biosensors closely depended on the Nd(3+) ion content. With increasing the Nd(3+) doping content, the electrochemical performance of the Hb-Chi-BNTO-x biosensors showed regularly variable. Moreover, compared with the Hb-Chi-BTO and other Hb-Chi-BNTO-x biosensors, the Hb-Chi-BNTO-0.85 biosensor had more excellent electrochemical and electrocatalytic properties such as stronger redox peak currents (approximately three-fold), smaller peak-to-peak separation (50 mV), larger heterogeneous electron transfer rate (14.1 ± 3.8s(-1)), higher surface concentration of electroactive redox protein (about 8.16 × 10(-11)mol/cm(2)), and better reproducibility and stability. The Nd-depended electrochemical properties of the Hb-Chi-BNTO biosensors may open up a new idea for designing third-generation electrochemical biosensors, and the BNTO-0.85-based biosensor is also expected to find potential applications in many areas such as biomedical, food, and environmental detection.     

Kinetic and biochemical properties of high and low redox potential laccases from fungal and plant origin

The electrochemical studies of laccase–mediator systems are aimed at understanding the mechanism of their redox transformation and their efficiency in both homogeneous and heterogeneous reactions; this topic has paramount application spanning from bleaching of paper pulp and the enzymatic degradation of lignin to the biosensors and biofuel cell development. In this paper four different laccases from Trametes hirsuta (ThL), Trametes versicolor (TvL), Melanocarpus albomyces (r-MaL) and Rhus vernicifera (RvL) were characterized from both biochemical and electrochemical points of view. Two of them (TvL and ThL) are high redox potential and two (RvL and r-MaL) are low redox potential laccases. The outline of this work is focused on the determination of catalytic and bioelectrochemical properties of these four enzymes in homogenous solution as well as immobilized onto electrode surface in the presence of a set of different redox mediators. The results measured in the homogenous reaction system correlated well with those measured with the immobilized enzymes. In addition, they are in good agreement with those reported with reference techniques, suggesting that the electrochemical methods employed in this work can be applied well in place of the traditional techniques commonly used for the kinetic characterization of laccases. These results are also discussed in terms of the known amino acid sequences and three-dimensional (3D) structures of the laccases.     

基于酶电极的乳酸检测生物传感器

乳酸(C₃H₆O₃),又名2-羟基丙酸、丙醇酸,属于羟基酸的一种。乳酸在食品工业、临床医学、生物技术等行业具有极其重要的意义,因此如何高通量检测不同样品中的乳酸成为目前业界研究的重点。传统乳酸检测方法操作繁琐、费时费力或需要昂贵的检测设备,乳酸生物传感器可以克服这些限制,不需要样品制备,能够快速、简便、可靠地定量测定食品或血浆中的乳酸,具有广阔的应用前景。乳酸酶电极生物传感器主要有两种类型——基于L-乳酸氧化酶(L-LOD)和L-乳酸脱氢酶(L-LDH)的乳酸生物传感器。本文综述了L-LOD和L-LDH结构特征、来源及催化机理,讨论了改善基于酶电极的乳酸传感器性能的3种策略(电极材料改造策略、酶固定化策略、酶分子工程改造策略),还根据用于制造乳酸生物传感器的不同载体包括膜、透明凝胶基质、水凝胶载体、纳米颗粒等对乳酸生物传感器进行了归类分析,最后本文将目前商品化应用的酶电极乳酸生物传感器特点进行了对比总结讨论,阐述了乳酸生物传感器的未来应用方向,并对未来发展前景进行了展望。     

DNA framework-engineered electrochemical biosensors

Self-assembled DNA nanostructures have shown remarkable potential in the engineering of biosensing interfaces, which can improve the performance of various biosensors. In particular, by exploiting the structural rigidity and programmability of the framework nucleic acids with high precision, molecular recognition on the electrochemical biosensing interface has been significantly enhanced, leading to the development of highly sensitive and specific biosensors for nucleic acids, small molecules,proteins, and cells. In this review, we summarize recent advances in DNA framework-engineered biosensing interfaces and the application of corresponding electrochemical biosensors.