Reusable surface confined semi-conducting metal-TCNQ and metal-TCNQF4 catalysts for electron transfer reactions

The synthesis of organic semiconducting materials based on silver and copper-TCNQ (TCNQ = 7,7,8,8-tetracyanoquinodimethane) and their fluorinated analogues has received a significant amount of attention due to their potential use in organic electronic applications. However, there is a scarcity in the identification of different applications for which these interesting materials may be suitable candidates. In this work, we address this by investigating the catalytic properties of such materials for the electron transfer reaction between ferricyanide and thiosulphate ions in aqueous solution, which to date has been almost solely limited to metallic nanomaterials. Significantly it was found that all the materials investigated, namely CuTCNQ, AgTCNQ, CuTCNQF4 and AgTCNQF4, were catalytically active and, interestingly, the fluorinated analogues were superior. AgTCNQF4 demonstrated the highest activity and was tested for its stability and re-usability for up to 50 cycles without degradation in performance. The catalytic reaction was monitored via UV-vis spectroscopy and open circuit potential versus time measurements, as well as an investigation of the transport properties of the films via electrochemical impedance spectroscopy. It is suggested that morphology and bulk conductivity are not the limiting factors, but rather the balance between the accumulated surface charge from electron injection via thiosulphate ions on the catalyst surface and transfer to the ferricyanide ions which controls the reaction rate. The facile fabrication of re-usable surface confined organic materials that are catalytically active may have important uses for many more electron transfer reactions.

Robust nanostructured silver and copper fabrics with localized surface plasmon resonance property for effective visible light induced reductive catalysis

Inspired by high porosity, absorbency, wettability and hierarchical ordering on the micrometer and nanometer scale of cotton fabrics, a facile strategy is developed to coat visible light active metal nanostructures of copper and silver on cotton fabric substrates. The fabrication of nanostructured Ag and Cu onto interwoven threads of a cotton fabric by electroless deposition creates metal nanostructures that show a localized surface plasmon resonance (LSPR) effect. The micro/nanoscale hierarchical ordering of the cotton fabrics allows access to catalytically active sites to participate in heterogeneous catalysis with high efficiency. The ability of metals to absorb visible light through LSPR further enhances the catalytic reaction rates under photoexcitation conditions. Understanding the mode of electron transfer during visible light illumination in Ag@Cotton and Cu@Cotton through electrochemical measurements provides mechanistic evidence on the influence of light in promoting electron transfer during heterogeneous catalysis for the first time. The outcomes presented in this work will be helpful in designing new multifunctional fabrics with the ability to absorb visible light and thereby enhance light-activated catalytic processes.

Proton translocation coupled to electron transfer reactions in terminal oxidases

Terminal oxidases are the final proteins of the respiratory chain in eukaryotes and some bacteria. They catalyze most of the biological oxygen consumption on Earth done by aerobic organisms. During the catalytic reaction terminal oxidases reduce dioxygen to water and use the energy released in this process to maintain the electrochemical proton gradient by functioning as a redox-driven proton pump. This membrane gradient of protons is extremely important for cells as it is used for many cellular processes, such as transportation of substrates and ATP synthesis. Even though the structures of several terminal oxidases are known, they are not sufficient in themselves to explain the molecular mechanism of proton pumping. In this work we have applied a complex approach using a variety of different techniques to address the properties and the mechanism of proton translocation by the terminal oxidases. The combination of direct measurements of pH changes during catalytic turnover, time-resolved potentiometric electrometry and optical spectroscopy, made it possible to obtain valuable information about various aspects of oxidase functioning. We compared oxygen binding properties of terminal oxidases from the distinct heme-copper (CcO) and cytochrome bd families and found that cytochrome bd has a high affinity for oxygen, which is 3 orders of magnitude higher than that of CcO. Interestingly, the difference between CcO and cytochrome bd is not only in higher affinity of the latter to oxygen, but also in the way that each of these enzymes traps oxygen during catalysis. CcO traps oxygen kinetically - the molecule of bound dioxygen is rapidly reduced before it can dissociate. Alternatively, cytochrome bd employs an alternative mechanism of oxygen trapping - part of the redox energy is invested into tight oxygen binding, and the price paid for this is the lack of proton pumping. A single cycle of oxygen reduction to water is characterized by translocation of four protons across the membrane. Our results make it possible to assign the pumping steps to discrete transitions of the catalytic cycle and indicate that during in vivo turnover of the oxidase these four protons are transferred, one at a time, during the P→F, F→OH, Oh→Eh, and Eh→R transitions. At the same time, each individual proton translocation step in the catalytic cycle is not just a single reaction catalyzed by CcO, but rather a complicated sequence of interdependent electron and proton transfers. We assume that each single proton translocation cycle of CcO is assured by internal proton transfer from the conserved Glu-278 to an as yet unidentified pump site above the hemes. Delivery of a proton to the pump site serves as a driving reaction that forces the proton translocation cycle to continue.

Oxidation of catechol in plants : III. Purification and properties of the diphenylene dioxide 2,3-quinone-forming enzyme system from Tecoma leaves

In attempting to determine the nature of the enzyme system mediating the conversion of catechol to diphenylenedioxide 2,3-quinone, in Tecoma leaves, further purification of the enzyme was undertaken. The crude enzyme from Tecoma leaves was processed further by protamine sulfate precipitation, positive adsorption on tricalcium phosphate gel, and elution and chromatography on DEAE-Sephadex. This procedure yielded a 120-fold purified enzyme which stoichiometrically converted catechol to diphenylenedioxide 2,3-quinone. The purity of the enzyme system was assessed by polyacrylamide gel electrophoresis. The approximate molecular weight of the enzyme was assessed as 200,000 by gel filtration on Sephadex G-150. The enzyme functioned optimally at pH 7.1 and at 35 °C. The Km for catechol was determined as 4 × 10−4 Image . The enzyme did not oxidize o-dihydric phenols other than catechol and it did not exhibit any activity toward monohydric and trihydric phenols and flavonoids. Copper-chelating agents did not inhibit the enzyme activity. Copper could not be detected in the purified enzyme preparations. The purified enzyme was not affected by extensive dialysis against copper-complexing agents. It did not show any peroxidase activity and it was not inhibited by catalase. Hydrogen peroxide formation could not be detected during the catalytic reaction. The enzymatic conversion of catechol to diphenylenedioxide 2,3-quinone by the purified Tecoma leaf enzyme was suppressed by such reducing agents as GSH and cysteamine. The purified enzyme was not sensitive to carbon monoxide. It was not inhibited by thiol inhibitors. The Tecoma leaf was found to be localized in the soluble fraction of the cell. Treatment of the purified enzyme with acid, alkali, and urea led to the progressive denaturation of the enzyme.

Endonuclease Active Site Plasticity Allows DNA Cleavage with Diverse Alkaline Earth and Transition Metal Ions

A majority of enzymes show a high degree of specificity toward a particular metal ion in their catalytic reaction. However, Type II restriction endonuclease (REase) R.KpnI, which is the first member of the HNH superfamily of REases, exhibits extraordinary diversity in metal ion dependent DNA cleavage. Several alkaline earth and transition group metal ions induce high fidelity and promiscuous cleavage or inhibition depending upon their concentration. The metal ions having different ionic radii and co-ordination geometries readily replace each other from the enzyme's active site, revealing its plasticity. Ability of R KpnI to cleave DNA with both alkaline earth and transition group metal ions having varied ionic radii could imply utilization of different catalytic site(s). However, mutation of the invariant His residue of the HNH motif caused abolition of the enzyme activity with all of the cofactors, indicating that the enzyme follows a single metal ion catalytic mechanism for DNA cleavage. Indispensability of His in nucleophile activation together with broad cofactor tolerance of the enzyme indicates electrostatic stabilization function of metal ions during catalysis. Nevertheless, a second metal ion is recruited at higher concentrations to either induce promiscuity or inhibit the DNA cleavage. Regulation of the endonuclease activity and fidelity by a second metal ion binding is a unique feature of R.KpnI among REases and HNH nucleases. The active site plasticity of R.KpnI opens up avenues for redesigning cofactor specificities and generation of mutants specific to a particular metal ion.

Splicing Functions and Global Dependency on Fission Yeast Slu7 Reveal Diversity in Spliceosome Assembly

The multiple short introns in Schizosaccharomyces pombe genes with degenerate cis sequences and atypically positioned polypyrimidine tracts make an interesting model to investigate canonical and alternative roles for conserved splicing factors. Here we report functions and interactions of the S. pombe slu7(+) (spslu7(+)) gene product, known from Saccharomyces cerevisiae and human in vitro reactions to assemble into spliceosomes after the first catalytic reaction and to dictate 3' splice site choice during the second reaction. By using a missense mutant of this essential S. pombe factor, we detected a range of global splicing derangements that were validated in assays for the splicing status of diverse candidate introns. We ascribe widespread, intron-specific SpSlu7 functions and have deduced several features, including the branch nucleotide-to-3' splice site distance, intron length, and the impact of its A/U content at the 5' end on the intron's dependence on SpSlu7. The data imply dynamic substrate-splicing factor relationships in multiintron transcripts. Interestingly, the unexpected early splicing arrest in spslu7-2 revealed a role before catalysis. We detected a salt-stable association with U5 snRNP and observed genetic interactions with spprp1(+), a homolog of human U5-102k factor. These observations together point to an altered recruitment and dependence on SpSlu7, suggesting its role in facilitating transitions that promote catalysis, and highlight the diversity in spliceosome assembly.

Splicing Functions and Global Dependency on Fission Yeast Slu7 Reveal Diversity in Spliceosome Assembly

The multiple short introns in Schizosaccharomyces pombe genes with degenerate cis sequences and atypically positioned polypyrimidine tracts make an interesting model to investigate canonical and alternative roles for conserved splicing factors. Here we report functions and interactions of the S. pombe slu7(+) (spslu7(+)) gene product, known from Saccharomyces cerevisiae and human in vitro reactions to assemble into spliceosomes after the first catalytic reaction and to dictate 3' splice site choice during the second reaction. By using a missense mutant of this essential S. pombe factor, we detected a range of global splicing derangements that were validated in assays for the splicing status of diverse candidate introns. We ascribe widespread, intron-specific SpSlu7 functions and have deduced several features, including the branch nucleotide-to-3' splice site distance, intron length, and the impact of its A/U content at the 5' end on the intron's dependence on SpSlu7. The data imply dynamic substrate-splicing factor relationships in multiintron transcripts. Interestingly, the unexpected early splicing arrest in spslu7-2 revealed a role before catalysis. We detected a salt-stable association with U5 snRNP and observed genetic interactions with spprp1(+), a homolog of human U5-102k factor. These observations together point to an altered recruitment and dependence on SpSlu7, suggesting its role in facilitating transitions that promote catalysis, and highlight the diversity in spliceosome assembly.

Developing acetylcholinesterase-based inhibition assay by modulated synthesis of silver nanoparticles: applications for sensing of organophosphorus pesticides

A novel and highly sensitive sensing strategy for the detection of organophosphorus compounds (OPs) based on the catalytic reaction of acetylcholinesterase (AChE) and acetylcholine (ATCh) during the modulated synthesis of silver nanoparticles (AgNPs) has been developed. The enzymatic hydrolysis of ATCh by AChE yields thiocholine (TCh), which induces the aggregation of AgNPs during synthesis, and the absorption peak at 382 nm corresponding to AgNPs decreases. The enzymatic reaction can be regulated by OPs, which can covalently bind to the active site of AChE and decrease the TCh formation, thereby decreasing the aggregation and significantly enhancing the absorption peak at 382 nm. The proposed system achieved good linearity and limits of detection of 0.078 nM and 2.402 nM for trichlorfon and malathion, respectively, by UV-visible spectroscopy. Further, the sensitivity of the proposed system was demonstrated through the determination of OPs in different spiked real samples. The described work shows the potential application for further development of a colorimetric sensor for other OP pesticide detection during the synthesis of AgNPs using enzyme-based assays.

Criação de mesoporos em zeólitas ZSM-5, mordenita e ferrierita e análise de seu efeito na reação de hidroisomerização do n-heptano

As zeólitas têm recebido grande atenção acadêmica e industrial devido às suas características ácidas e estruturais. A estrutura da zeólita pode ser utilizada para conduzir uma reação catalítica na direção do produto desejado, evitando assim reações paralelas. Porém, essa mesma estrutura cria restrições difusivas com relação ao acesso aos sítios ativos no interior dos microporos. Neste trabalho foram estudados dois métodos de criação de mesoporos (térmico e básico) com o intuito de modificar a acessibilidade aos sítios catalíticos das zeólitas. A reação de hidroisomerização do n-heptano foi selecionada para avaliar as zeólitas após a criação de mesoporosidade. O tratamento térmico (via calcinação em temperaturas elevadas) foi utilizado para as zeólitas do tipo ZSM-5, Mordenita e Ferrierita, tendo sido observado um aumento pouco significativo na mesoporosidade. Este tratamento promoveu, porém, uma significativa desaluminização das amostras, acompanhada da formação de quantidades importantes de espécies de Al extra-rede (ALER), o que se refletiu num bloqueio parcial dos mesoporos gerados e dos microporos preexistentes, e na redução na densidade de sítios ácidos das amostras. A ampliação da escala do tratamento térmico (aumento da quantidade tratada de 2 g para 30 g) não se mostrou reprodutível, gerando menos mesoporos do que o observado no preparo em pequena escala. O tratamento básico (via dessilicação por meio de NaOH), ao contrário do anterior, promoveu a formação de mesoporos gerando menos quantidade de ALER e se mostrou mais reprodutível quando da ampliação da escala. O desempenho dos catalisadores Pt/Al2O3+zeólita na reação de hidroisomerização do n-heptano foi influenciado pela densidade de sítios ácidos fortes e pela estrutura porosa da zeólita. Com relação ao efeito dos tratamentos térmico e básico sobre o desempenho dos catalisadores à base de ZSM-5, os resultados mostraram que o comportamento do catalisador submetido ao tratamento básico (Pt/Al2O3+BZSM-5/85-2) foi similar ao do tratado termicamente (Pt/Al2O3+TZSM-5/1000-2) com relação à distribuição de produtos na reação de hidroisomerização do n-heptano, particularmente com relação aos produtos leves e aos isômeros monorramificados. No entanto, a presença mais significativa de mesoporos na zeólita após tratamento básico (BZSM-5/85-2), se refletiu num leve favorecimento à formação dos isômeros birramificados

Synthesis and characterization of Prussian blue@platinum nanoparticle hybrids from a mixture solution of platinum nanocatalyst and ferric ferricyanide

In the present study, platinum nanoparticles modified with Prussian blue (PB) have been synthesized by a heterogeneous catalytic reaction. Transmission electronic microscopy (TEM) confirmed the deposition of nanoclusters around the Surfaces of platinum particles, and spectroscopic studies verified that the molecular composition of the nanoclusters was dominantly PB and a minority of platinum ferricyanide. Thus, it was shown that the platinum particles behaved not only as catalysts for the growth of PB, but also as a reactant to generate a PB analogue complex.

The electrocatalytic oxidation of ascorbic acid on polyaniline film synthesized in the presence of camphorsulfonic acid

Polyaniline-camphorsulfonic acid (PAN-CSA) composite film on platinum electrode surface has been synthesized via the electrochemical polymerization of aniline in the presence of camphorsulfonic acid (CSA). It was found that the doping of polyaniline (PAN) with CSA extends the electroactivity of PAN in neutral and even in alkaline media. The PAN-CSA composite film coated platinum electrodes are shown to be good electrocatalytic surfaces for the oxidation of ascorbic acid (AA) in phosphate buffer solution (PBS) of pH 7.0. The anodic peak potential of AA shifts from 0.63 V at the bare platinum electrode to 0.34 V at the PAN-CSA composite modified platinum electrode with a greatly enhanced current response. A linear calibration graph is obtained over the AA concentration range of 5-50 mM using cyclic voltammetry. The kinetics of the catalytic reaction are investigated using rotating disk electrode voltammetry and chronoamperometry. The results are explained using the theory of electrocatalytic reactions at chemically modified electrodes. The PAN-CSA composite on the electrode surface shows good reproducibility and stability.