Electrochemical characteristics of glucose oxidase adsorbed at carbon nanotubes modified electrode with ionic liquid as binder

Multiwall carbon nanotubes (CNTs)-modified electrode has been prepared by using ionic liquid (IL) as the binder. The as-prepared CNTs-IL composite modified electrode has good biocompatibility and is a suitable matrix to immobilize biomolecules. Glucose oxidase (GOx), containing flavin adenine dinucleotide as active site, stably adsorbed on modified electrode surface has resulted in the direct electron transfer. The electron transfer rate of 9.08 s(-1) obtained is much higher than that of GOx adsorbed on the CNTs papers (1.7 s(-1)), and the process is more reversible with small redox peak separation of 23 mV This may be due to the synergetic promotion of CNTs and IL to electron transfer of the protein, especially the IL as the binder, showing better electrochemical properties than that of chitosan and Nafion. Furthermore, GOx adsorbed at the modified electrode exhibits good stability and keeps good electrocatalytic activity to glucose with broad linear range up to 20 mM. Besides, the simple preparation procedure and easy renewability make the system a basis to investigate the electron transfer kinetics and biocatalytic performance of GOx and provide a promising platform for the development of biosensors.

Study on the lifetime of M-intermediate of bacteriorhodopsin film chemically modified with CeO2 nanoparticles

The characteristics of intermediates of bacteriorhodopsin (bR) can be verified by chemical modification of its surroundings. CeO2 nanoparticles, which were obtained using water-in-oil (W/O) microemulsion and calcined at various temperatures, were used as chemical additive for the modification of bR. X-ray diffraction (XRD) shows that the mean particle sizes for the samples calcined at 500 and 800 degrees C are approximately 10 and 30 nm, respectively. We prepared CeO2 nanoparticle modified poly(vinyl alcohol) (bR-PVA) films with an optical density of about 1.5 at the ground state. It is observed that the lifetime of the Wintermediate for the modified films is prolonged compared with that of the unmodified ones, and the lifetime increases with decreasing particle size. A probable mechanism, which is likely to involve effective molecular interactions between the CeO2 nanoparticles and the bR molecules, is discussed. The hydroxyl groups, which might arise from the interaction between the nanoparticles and the surrounding water molecules, help to lower the ability of the Schiff base of uptaking protons in the Wintermediate. The results indicate that controlling the interactions between biomolecules and various nanomaterials would enlarge the functionality and the range of the application of nanoparticles.

Carbon nanotubes and glucose oxidase bionanocomposite bridged by ionic liquid-like unit: Preparation and electrochemical properties

For their biocompatibility and potential bionanoelectronic applications, integration of carbon nanotubes (CNTs) with biomolecules such as redox enzyme is highly anticipated. Therein, CNTs are expected to act not only as an electron transfer promoter, but also as immobilizing substrate for biomolecules. In this report, a novel method for immobilization of biomolecules on CNTs was proposed based on ionic interaction, which is of universality and widespread use in biological system. As illustrated, glucose oxidase (GOD) and single-walled carbon nanotubes (SWNTs) were integrated into a unitary bionanocomposite by means of ionic liquid-like unit on functionalized SWNTs. The resulted bionanocomposite illustrated better redox response of immobilized GOD in comparison of that prepared by weak physical absorption without ionic interaction. As a potential application of concept, the electrochemical detection of glucose was exemplified based on this novel bionanocomposite.

Quantifying the kinetic paths of flexible biomolecular recognition

Biomolecular recognition often involves large conformational changes, sometimes even local unfolding. The identification of kinetic pathways has become a central issue in understanding the nature of binding. A new approach is proposed here to study the dynamics of this binding-folding process through the establishment of a path-integral framework on the underlying energy landscape. The dominant kinetic paths of binding and folding can be determined and quantified. The significant coupling between the binding and folding of biomolecules often exists in many important cellular processes. In this case, the corresponding kinetic paths of binding are shown to be intimately correlated with those of folding and the dynamics becomes quite cooperative. This implies that binding and folding happen concurrently. When the coupling between binding and folding is weak (strong), the kinetic process usually starts with significant folding (binding) first, with the binding (folding) later proceeding to the end. The kinetic rate can be obtained through the contributions from the dominant paths. The rate is shown to have a bell-shaped dependence on temperature in the concentration-saturated regime consistent with experiment. The changes of the kinetics that occur upon changing the parameters of the underlying binding-folding energy landscape are studied.

Electrochemical biosensors based on advanced bioimmobilization matrices

Biosensors have experienced rapid, extensive development. To maintain the bioactivity of biomolecules and to give the electrochemical output signal required, appropriate bioimmobilization matrices for biomolecules are critical.In this review, we describe some advanced membrane materials (including hydrogels, sol-gel-derived organic-inorganic composites and lipid membranes), introduce electrochemical biosensors based on bioimmobilization materials and describe their performance.Biosensors operating in extreme conditions and displaying direct electron transfer with electrodes based on these advanced membrane materials are attractive. Recent developments in nanomaterials include biosensors, so we emphasize the intersection of nanomaterials with advanced membrane materials in biosensors.

Electrochemical characteristics of facile prepared carbon nanotubes-ionic liquid gel modified microelectrode and application in bioelectrochemistry

The carbon nanotubes (CNTs) based microelectrode (ME) by modifying CNTs-room temperature ionic liquid (IL) gel at carbon fiber microelectrode (CFME) is easily prepared, which exhibits the typical cyclic voltammogram of ME with sigmoid shape and possesses good stability, high conductivity and enlarged current response and tunable dimension. The direct electron transfer of glucose oxidase has been greatly promoted showing reversible electrochemical behavior even at high scan rate. In addition, the CNTs based ME also exhibits effectively electrocatalytic oxidized ability to biomolecules, e.g. dopamine (DA), ascorbic acid (AA) and dihydronicotinamide adenine dinucleotide. The obvious separation of oxidized peak potential for DA and AA makes it possible to selectively determine DA in presence of AA. These phenomena show that the CNTs based ME has promising potential to detect various species in vivo and in vitro.

Manipulation, dissection, and lithography using modified tapping mode atomic force microscope

A modified tapping mode of the atomic force microscope (AFM) was introduced for manipulation, dissection, and lithography. By sufficiently decreasing the amplitude of AFM tip in the normal tapping mode and adjusting the setpoint, the tip-sample interaction can be efficiently controlled. This modified tapping mode has some characteristics of the AFM contact mode and can be used to manipulate nanoparticles, dissect biomolecules, and make lithographs on various surfaces. This method did not need any additional equipment and it can be applied to any AFM system.

Single-molecule dynamics reveals cooperative binding-folding in protein recognition

The study of associations between two biomolecules is the key to understanding molecular function and recognition. Molecular function is often thought to be determined by underlying structures. Here, combining a single-molecule study of protein binding with an energy-landscape-inspired microscopic model, we found strong evidence that biomolecular recognition is determined by flexibilities in addition to structures. Our model is based on coarse-grained molecular dynamics on the residue level with the energy function biased toward the native binding structure ( the Go model). With our model, the underlying free-energy landscape of the binding can be explored. There are two distinct conformational states at the free-energy minimum, one with partial folding of CBD itself and significant interface binding of CBD to Cdc42, and the other with native folding of CBD itself and native interface binding of CBD to Cdc42. This shows that the binding process proceeds with a significant interface binding of CBD with Cdc42 first, without a complete folding of CBD itself, and that binding and folding are then coupled to reach the native binding state.

The electrochemistry of cytochrome c at a viologen-thiol self-assembled monolayer

At the self-assembled monolayer (SAM) of a thiol-functionalized viologen modified gold electrode, cytochrome c (cyt c) exhibits a quasi-reversible electrochemical reaction. The heterogeneous electron transfer rate constant of cyt c in 0.1 mol/L phosphate buffer solution(pH 6.96) is 0.164 cm.s(-1) at 500 mV/s. The adsorbed cyt c on the viologen SAM forms a closely packed monolayer, whose average electron transfer rate is 4.85 s(-1) in the scan range of 50 to 500 mV/s. These results suggest that the SAM of viologen-thiol is a relatively stable, ordered and well-behaved monolayer from an electrochemical standpoint and it promotes the electron transfer process of biomolecules on electrode surface well.

A COMPARATIVE-STUDY ON STM IMAGING AND ELECTROCATALYTIC ACTIVITY OF DIFFERENT SURFACES MODIFIED WITH FLAVIN ADENINE-DINUCLEOTIDE

Flavin adenine dinucleotide (FAD) was modified onto the highly oriented pyrolytic graphite (hopg) and glassy carbon electrode (gee) surfaces with three methods, respectively. Corresponding image analysis for FAD-modified hopg surfaces has been performed by scanning tunnelling microscope (STM) for the first time. The molecular resolution STM image of FAD adsorbed on the freshly-cleaved hopg was obtained, the quantitative size determination suggests that the FAD molecules adsorb side lying on the substrate surface. The anodization treatment of hopg surface yields many pits, which were clearly observed under STM. These pits provide active sites on the hopg surface for modification and the treated hopg can strongly adsorb FAD molecules, the latter exhibiting an irregular cluster structure on such a surface. When FAD was electrochemically deposited on the substrate surface, a chain structure was successfully observed. The adsorbed FAD on anodized glassy carbon electrode (gee) surface can effectively catalyze the reduction of glucose oxidase, hemoglobin and myoglobin, with a large decrease in the overvoltage, whereas the deposited FAD film exhibits excellent electrocatalysis towards dioxygen reduction.

Spectral changes of C-phycocyanin with different molar ratios of SPDP

Pure C-phycocyanin was prepared from Spirulina platensis using one-step anion-exchange chromatography. The C-PC obtained was with an absorption maximum at 620 nm and a fluorescence emission maximum at 640 nm when excited by 580 nm. SPDP is an excellent heterobifunctional crosslinker for thiolating amines. Different molar ratios of SPDP have remarkable influence on the absorption and fluorescence spectra of C-phycocyanin. The absorption maximum and fluorescence emission maximum both decreased and blue-shifted from 640 run to 630 nm as the molar ratios of SPDP increased. It was found that the molar ratios of SPDP to C-phycocyanin was not more than 100 was appropriate to being conjugated with other biomolecules from the absorption and fluorescence spectra of C-phycocyanin.

The unique skeleton of siliceous sponges (Porifera; Hexactinellida and Demospongiae) that evolved first from the Urmetazoa during the Proterozoic: a review

Sponges (phylum Porifera) had been considered as an enigmatic phylum, prior to the analysis of their genetic repertoire/tool kit. Already with the isolation of the first adhesion molecule, galectin, it became clear that the sequences of sponge cell surface receptors and of molecules forming the intracellular signal transduction pathways triggered by them, share high similarity with those identified in other metazoan phyla. These studies demonstrated that all metazoan phyla, including Porifera, originate from one common ancestor, the Urmetazoa. The sponges evolved prior to the Ediacaran-Cambrian boundary (542 million years ago [myr]) during two major "snowball earth events", the Sturtian glaciation (710 to 680 myr) and the Varanger-Marinoan ice ages (605 to 585 myr). During this period the ocean was richer in silica due to the silicate weathering. The oldest sponge fossils (Hexactinellida) have been described from Australia, China and Mongolia and are thought to have existed coeval with the diverse Ediacara fauna. Only little younger are the fossils discovered in the Sansha section in Hunan (Early Cambrian; China). It has been proposed that only the sponges possessed the genetic repertoire to cope with the adverse conditions, e.g. temperature-protection molecules or proteins protecting them against ultraviolet radiation. The skeletal elements of the Hexactinellida (model organisms Monorhaphis chuni and Monorhaphis intermedia or Hyalonema sieboldi) and Demospongiae (models Suberites domuncula and Geodia cydonium), the spicules, are formed enzymatically by the anabolic enzyme silicatein and the catabolic enzyme silicase. Both, the spicules of Hexactinellida and of Demospongiae, comprise a central axial canal and an axial filament which harbors the silicatein. After intracellular formation of the first lamella around the channel and the subsequent extracellular apposition of further lamellae the spicules are completed in a net formed of collagen fibers. The data summarized here substantiate that with the finding of silicatein a new aera in the field of bio/inorganic chemistry started. For the first time strategies could be formulated and experimentally proven that allow the formation/synthesis of inorganic structures by organic molecules. These findings are not only of importance for the further understanding of basic pathways in the body plan formation of sponges but also of eminent importance for applied/commercial processes in a sustainable use of biomolecules for novel bio/inorganic materials.

Optymalizacja metod analizy Főrsterowskiego rezonansowego przeniesienia energii (FRET) pojedynczych cząsteczek

Wydział Chemii: Zakład Chemii Fizycznej

Synthesis of iminophosphine and phosphinoiminol cyclometallated Pt (II) and Pt (IV) chloro complexes and studies into their biological and photophysical properties

This thesis focuses on the synthesis and analysis of novel chloride based platinum complexes derived from iminophosphine and phosphinoamide ligands, along with studies on their reactivity towards substitution and oxidation reactions. Also explored here are the potential applications of these complexes for biological and luminescent purposes. Chapter one provides an extensive overview of platinum coordination chemistry with examples of various mixed donor ligands along with the history of platinum anticancer therapy. It also looks at metals in medicine, both for biological functions as well as for therapeutic purposes and gives a background to some other applications for platinum complexes. Chapter two outlines the design and synthetic strategies employed for the development of novel platinum (II) chloride complexes from iminophosphine and phosphinoamide ligands. Also reported is the cyclometallation of these complexes to form stable tridentate mixed donor platinum (II) compounds. In Chapter three the development of a direct method for displacing a chloride from a platinum metal centre with a desired phosphine is reported. Numerous methods for successful oxidation of the platinum (II) complexes will also be explored, leading to novel platinum (IV) complexes being reported here also. The importance of stabilisation of the displaced anion, chloride, by the solvent system will also be discussed in this chapter. Chapter four investigates the reactivity of the platinum (II) complexes towards two different biomolecules to form novel platinum bio-adducts. The potential application of the platinum (II) cyclometallates as chemotherapeutics will also be explored here using in-vitro cancer cell testing. Finally, luminescence studies are also reported here for the ligands and platinum complexes reported in chapter two and three to investigate potential applications in this field also. Chapter five provides a final conclusion and an overall summary of the entire project as well as identifying key areas for future work.