Enzyme microgels in packed-bed bioreactors with downstream amperometric detection using microfabricated interdigitated microsensor electrode arrays.

In this article, we describe the use of pH- responsive hydrogels as matrices for the immobilization of two enzymes, glucose oxidase (GOx) and glutamate oxidase (GlutOx). Spherical hydrogel beads were prepared by inverse suspension polymerization and the enzymes were immobilized by either physical entrapment or covalent immobilization within or on the hydrogel surface. Packed-bed bioreactors were prepared containing the bioactive hydrogels and these incorporated into flow injection (FI) systems for the quantitation of glucose and monosodium glutamate (MSG) respectively. The FI amperometric detector comprised a microfabricated interdigitated array within a thin-layer flow cell. For the FI manifold incorporating immobilized GOx, glucose response curves were found to be linear over the concentration range 1.8-280 mg dL(-1) (0.1-15.5 mM) with a detection limit of 1.4 mg dL(-1) (0.08 mM). Up to 20 samples can be manually analyzed per hour, with the hydrogel-GOx bioreactor exhibiting good within-day (0.19%) precision. The optimized FI manifold for MSG quantitation yielded a linear response range of up to 135 mg dL(-1) (8 mM) with a detection limit of 3.38 mg dL(-1) (0.2 mM) and a throughput of 30 samples h(-1). Analysis of commercially produced soup samples gave a within-day precision of 3.6%. Bioreactors containing these two physically entrapped enzymes retained > 60% of their initial activities after a storage period of up to 1 year.     

A Study of Protein Electrochemistry on a Supported Membrane Electrode

Protein electrochemistry offers a direct method to identify and characterize biological electron transfer processes, potentially leading to commercial applications such as biosensors and diagnostic tools. However, establishing a biocompatible electrode interface that maintains the native state of the redox protein involves several challenges. In general, membrane proteins require the presence of a phospholipid bilayer to maintain their biological activity. Synthetic `biomimetic’ membranes are widely used to characterize membrane proteins, however they have seldom been applied to measurements of protein redox activity in electrochemical cells due to their inherent insulating property. In this study we demonstrate the use of the phospholipids: PC, PC/PG and PC/PG/cholesterol membrane mixtures on chemically modified (supported) gold electrode surfaces for direct protein electrochemistry. We compare the electrochemical activity of a relatively small, redox active “test protein”, cytochrome c, in the presence and absence of phospholipid on a gold electrode modified with thiol self assembled monolayers, to explore the effect of chain length and composition of the thiol on the charge coupling. Three thiols were investigated as self assembled monolayers on a gold electrode: octanethiol, mercaptopropionic and mercaptoundecanoic acid. We demonstrate here that the charge transfer efficiency of cytochrome c is better in the presence of the membrane and in addition, a superior redox response is obtained with surfaces modified with a thiol functionalised with a carboxylic acid.On leave from: Research Group on Laser Physics of the Hungarian Academy of Sciences, University of Szeged, Szeged, Hungary.Australian Peptide Conference Issue.     

Influence of chlorophyll concentrations on the reliability of measurements with an oxygen electrode when studying photosynthetic efficiency in bioassays using microalgae

Euglena gracilis and Chlamydomonas reinhardtii were used as model organisms to establish the best conditions for studying photosynthetic efficiency using the Light Pipette – experimental system, which enables sensitive detection of changes in oxygen evolution/consumption and an easy collection and digitalisation of data. Chlorophyll concentrations of 0.005, 0.025, 0.050 and 0.075 mgmL^-1 were investigated using different light regimes. Cultures of E. gracilis at the same chlorophyll concentration absorbed more light(measured at 580 μmol m^-2 s^-1) than those of C. reinhardtii. Cell density had a considerable effect on the reliability of measurements. Chlorophyll concentrations between 0.025 mg_Chl ml^-1 and 0.050 mg_Chlml^-1 can be recommended when applying the Light Pipette system in bioassays using microalgae. This revised version was published online in August 2006 with corrections to the Cover Date.     

Root exudates from sunflower (Helianthus annuus L.) show a strong adsorption ability toward Cd(II)

Abstract

Cd(II) adsorption of root exudates from sunflower (Helianthus annuus L.) seedling was investigated by Cd ion-selective electrode, Fourier Transform Infrared spectroscopy, and fluorescence spectroscopy. Root exudates from Helianthus annuus L. had strong adsorption ability toward Cd(II). The adsorption process was pH-dependent and the maximum adsorption capacity, 150.8 mg g^?1, was observed at pH 7.0. Root exudates had pK _a1 at 4.7 for carboxyl and pK _a2 at 9.2 for phenolic, and amino groups. The aliphatic and aromatic (C?H) groups, amide III group, and the C (=O)?O and sulfonate groups were responsible for Cd(II) adsorption. The excitation emission matrix fluorescence spectroscopy showed protein-like substances participated in Cd adsorption and formed strong complexes, with conditional stability constants of 4.70 and 4.32, which is a little lower than that determined by potentiometric methods, 5.13. The strong Cd complexing ability of root exudates implies that root exudates may significantly affect mobility, toxicity, and phytoavailability of Cd. Cd binding of root exudates may be attributed to its interaction with the proteins, polysaccharides, and phenolic compounds in root exudates.     

A Material Perspective of Rechargeable Metallic Lithium Anodes

Rechargeable lithium‐based batteries are long considered as the most promising candidates for application in various electronic devices, electric vehicles, and even electrical grids owing to their ultrahigh energy densities. However, to date, metallic lithium‐based batteries are still far from practical applications due to the low Coulombic efficiency and fast capacity decay of lithium anodes. The poor electrochemical performances of metallic lithium anodes are inherently related to random growth of lithium dendrites and infinite volume charge of lithium anodes. In this review, the failure mechanisms of metallic lithium anodes are summarized and ascribed to the unstable and inhomogeneous solid electrolyte interphase, uneven distributions of electric field, and lithium‐ion flux during the lithium plating processes. Correspondingly, efficient strategies for mitigating these problems, including surficial engineering, electric field, and lithium‐ion flux regulation are discussed from the perspective of anode materials. Finally, an outlook is proposed for the design and fabrication of next‐generation rechargeable metallic lithium anodes that aims to address the intrinsic problems of metallic lithium anodes.     

Mechanical Property Evolution of Silicon Composite Electrodes Studied by Environmental Nanoindentation

Mechanical degradation is largely responsible for the short cycle life of silicon (Si)‐based electrodes for future lithium‐ion batteries. An improved fundamental understanding of the mechanical behavior of Si electrodes, which evolves, as demonstrated in this paper, with the state of charge (SOC) and the cycle number, is a prerequisite for overcoming mechanical degradation and designing high capacity and durable Si‐based electrodes. In this study, Young's modulus (E) and hardness (H) of Si composite electrodes at different SOCs and after different cycle numbers are measured by nanoindentation under both dry and wet (liquid electrolyte) conditions. Unlike electrodes made of Si alone, E and H values of Si composite electrodes increase with increasing Li concentration. The composite electrodes under wet conditions are softer than that under dry conditions. Both E and H decrease with the cycle number. These findings highlight the effects of porosity, liquid environment, and degradation on the mechanical behavior of composite electrodes. The methods and results of this study on the mechanical property evolution of Si/polyvinylidene fluoride electrodes form a basis for exploring more effective binders for Si‐based electrodes. Furthermore, the evolving nature of the mechanical behavior of composite electrodes should be taken into consideration in future modeling efforts of porous composite electrodes.     

对吸附式电极记录装置的技术改进

本文介绍一种用吸附式电极记录合体细胞组织生物电位的改进装置。它的特点是在建造负压的注射器和吸附式电极之间设置一分离的小室。这一小室既保障了放大器与实验标本之间的电路连系,又可直接放置在实验标本附近,负压由改进的注射器经过充有空气的塑料管抽吸,注射器可以放在任何方便的位置上。该方法在记录小动物,如蜗牛、青蛙、等的心脏、消化道等组织器官的生物电位时都能获得比较理想的效果,对研究小动物合体细胞组织的正常机能及药物作用等都具有较好的适用价值,并具有定位准确、操作方便的优点。     

Potential‐resolved electrochemiluminescence of Ru(bpy)32+/C2O42− system on gold electrode

The electrogenerated chemiluminescence of Ru(bpy)₃²⁺/C₂O₄^2? system on a pre‐polarized Au electrode was studied using a potential‐resolved electrochemiluminescence (PRECL) method. Two anodic ECL peaks were observed at 1.22 V (vs. SCE) (EP1), 1.41 V (vs. SCE) (EP2), respectively. The effects of the concentration of oxalate and Ru(bpy)₃²⁺, adsorbed sulphur, CO₂, O₂, pH of the solution and pretreatment of the Au electrode on the two PRECL peaks were examined. The surface state of the pre‐oxidized gold electrode was also studied using the X‐ray photoelectron spectroscopy (XPS) technique. Moreover, comparative studies on i–E and I–E curves were carried out and a possible mechanism involving both the catalytic and the direct electro‐oxidation pathways was proposed for the ECL of Ru(bpy)₃²⁺/C₂O₄^2? system. EP1 is attributed to the Ru(bpy)₃^2/3+ reaction catalysed by C₂O₄^2? to generate Ru(bpy)₃^2+*. EP2 is likely because C₂O₄^2? was oxidized at the electrode to form CO₂^?·, followed by reaction with Ru(bpy)₃³⁺ to generate Ru(bpy)₃^2+*. Copyright © 2002 John Wiley & Sons, Ltd.     

Measuring the pH dependence of hydrogenase activities

The pH dependences of activities of homogenous hydrogenases of Thiocapsa roseopersicina and Desulfomicrobium baculatum in the reaction of hydrogen uptake in solution in the presence of benzyl viologen and the pH dependences of catalytic currents of hydrogen oxidation by electrodes on which these hydrogenases were immobilized were compared. Maximal activities of the hydrogenases from T. roseopersicina and D. baculatum in the reaction hydrogen uptake in solution were observed at pH 9.5 and 8.5, respectively. However, the steady-state current caused by catalytic uptake of hydrogen was maximal for the T. roseopersicina hydrogenase-containing electrode at pH 5.5-6.5 under overvoltage of 30-60 mV, whereas for electrodes with D. baculatum hydrogenase it was maximal at pH 6.0-6.5. Analysis of these data suggests that pH-dependent changes in the hydrogenase activities in solution during hydrogen uptake are due not only to the effect of proton concentration on the enzyme conformation or protonation of certain groups of the enzyme active center, but they are rather indicative of changes in free energy of the reaction accompanying changes in pH.     

Chromatic photoacclimation, photosynthetic electron transport and oxygen evolution in the Chlorophyll d-containing oxyphotobacterium Acaryochloris marina

Changes in photosynthetic pigment ratios showed that the Chlorophyll d-dominated oxyphotobacterium Acaryochloris marina was able to photoacclimate to different light regimes. Chl d per cell were higher in cultures grown under low irradiance and red or green light compared to those found when grown under high white light, but phycocyanin/Chl d and carotenoid/Chl d indices under the corresponding conditions were lower. Chl a, considered an accessory pigment in this organism, decreased respective to Chl d in low irradiance and low intensity non-white light sources. Blue diode PAM (Pulse Amplitude Modulation) fluorometry was able to be used to measure photosynthesis in Acaryochloris. Light response curves for Acaryochloris were created using both PAM and O₂ electrode. A linear relationship was found between electron transport rate (ETR), measured using a PAM fluorometer, and oxygen evolution (net and gross photosynthesis). Gross photosynthesis and ETR were directly proportional to one another. The optimum light for white light (quartz halogen) was about 206 ± 51 μmol m^− 2 s^− 1 (PAR) (Photosynthetically Active Radiation), whereas for red light (red diodes) the optimum light was lower (109 ± 27 μmol m^− 2 s^− 1 (PAR)). The maximum mean gross photosynthetic rate of Acaryochloris was 73 ± 7 μmol mg Chl d^− 1 h^− 1. The gross photosynthesis/respiration ratio (P_g/R) of Acaryochloris under optimum conditions was about 4.02 ± 1.69. The implications of our findings will be discussed in relation to how photosynthesis is regulated in Acaryochloris.