Functional Expression of a Mo-Cu-Dependent Carbon Monoxide Dehydrogenase (CODH) and Its Use as a Dissolved CO Bio-microsensor.

Herein, we report the heterologous expression in of a Mo-Cu-containing carbon monoxide dehydrogenase (Mo-Cu CODH) from , which resulted in an active protein catalyzing CO oxidation to CO. By supplying the growth medium with NaMoO (Mo) and CuSO (Cu), the Mo-Cu CODH metal cofactors precursors, the expressed L-subunit was found to have CO-oxidation activity even without the M- and S- subunits. This successful expression of CO-oxidizing-capable single L-subunit provides direct evidence of its role as the catalytic center of Mo-Cu CODH that has not been discovered and studied before.

Urea Biosensor Based on a CO Microsensor.

Urea sensors based on electrodes in direct contact with the medium have limited long-term stability when exposed to complex media. Here, we present a urea biosensor based on urease immobilized in an alginate polymer, buffered at pH 6, and placed in front of a newly developed fast and sensitive CO microsensor, where the electrodes are shielded by a gas-permeable membrane. The CO produced by the urease in the presence of urea diffuses into the microsensor and is reduced at a Ag cathode.

Ion Selective Amperometric Biosensors for Environmental Analysis of Nitrate, Nitrite and Sulfate.

Inorganic ions that can be redox-transformed by living cells can be sensed by biosensors, where the redox transformation gives rise to a current in a measuring circuit. Such biosensors may be based on enzymes, or they may be based on application of whole cells. In this review focus will be on biosensors for the environmentally important ions NO, NO, and SO, and for comparison alternative sensor-based detection will also be mentioned.

Fast Responding Amperometric CO Microsensor with Ionic Liquid-Aprotic Solvent Electrolytes.

Knowledge about the microscale distribution of CO is essential in many environmental and technical settings, and electrochemical CO sensing may be optimized to yield such information. The performance of a Clark-type CO sensor was greatly improved by adding 20% dimethylformamide (DMF) to the ionic liquid 1-ethyl-3-methylimidazolium dicyanamide (EMIM-DCA) previously used as an electrolyte. The addition of DMF resulted in a much faster response to increasing (95% response of about 100 s) or decreasing CO concentration, a negligible interference from low concentrations of NO, and a signal temperature dependence similar to that of O microsensors.

Controlling Voltage Reversal in Microbial Fuel Cells.

Microbial fuel cell (MFC) systems have been developed for potential use as power sources, along with several other applications, with bacteria as the prime factor enabling electrocatalytic activity. Limited voltage and current production from unit cells limit their practical applicability, so stacking multiple MFCs has been proposed as a way to increase power production. Special attention is paid to voltage reversal (VR), a common occurrence in stacked MFCs, and to identifying the mechanisms underlying this phenomenon.

Electrochemical Enzyme-Linked Sandwich Assay with a Cellulase Label for Ultrasensitive Analysis of Synthetic DNA and Cell-Isolated RNA.

Electrochemical enzyme-linked sandwich assays on magnetic beads (MBs) refer to one of the most sensitive approaches for analysis of nonamplified nucleic acid samples, with redox enzymes being routinely used as labels. Here, we report a sensitive and inexpensive electrochemical nucleic acid sandwich assay on MBs that exploits a hydrolytic enzyme cellulase as a label, while MBs are used for preconcentration and bioseparation of analyzed samples. Binding of target DNA or RNA to capture DNA-modified MB triggers sandwich assembly and its labeling with cellulase.

Electrochemical Assay for a Total Cellulase Activity with Improved Sensitivity.

Electrochemical methods allow fast and inexpensive analysis of enzymatic activities. Here, we report a simple and yet efficient electrochemical assay for the total activity of cellulase, a hydrolytic enzyme widely used in food and textiles industries, and for production of bioethanol. The assay exploits the difference in electrochemical signals from a soluble redox indicator, ferricyanide, on nitrocellulose films treated by cellulases.

Functionalized Polyacrylonitrile Nanofibrous Membranes for Covalent Immobilization of Glucose Oxidase.

Nanofibrous membrane (NFM) with uniform morphology and large surface area was prepared from 10% solution of polyacrylonitrile (PAN) in N,N-dimethylformamide by electrospinning technique. NFM was chemically modified for use as a support for the immobilization of glucose oxidase. Chemical modification of NFM was carried out by two different methods.

Graphitized-Carbon-Nanofiber Paper-Enzyme Electrode Fabrication Through Non-Covalent Modification for Enzyme Biofuel Cell Application.

Carbon nanofibers are an emerging smart material that are promising for use as a biosensor and a biofuel cell transducer material due to their morphological and electrochemical characteristics. In particular, graphitized carbon nanofibers possess unique structures of graphite-like edges within their high surface area that provide a large active site for enzyme attachment. For a specific application such as a biofuel cell, which requires highly stable electrical communication and electricity generation, non-covalent enzyme immobilization using bifunctional molecule is suggested as an appropriate approach because it does not change the carbon hybridization from sp2 to sp3 as covalent immobilization by acid treatment does.

Bioelectronic platforms for optimal bio-anode of bio-electrochemical systems: From nano- to macro scopes.

The current trend of bio-electrochemical systems is to improve strategies related to their applicability and potential for scaling-up. To date, literature has suggested strategies, but the proposal of correlations between each research field remains insufficient. This review paper provides a correlation based on platform techniques, referred to as bio-electronics platforms (BEPs).

Gated electron transfer reactions of truncated hemoglobin from Bacillus subtilis differently orientated on SAM-modified electrodes.

Electron transfer (ET) reactions of truncated hemoglobin from Bacillus subtilis (trHb-Bs) are suggested to be implicated in biological redox signalling and actuating processes that may be used in artificial environment-sensing bioelectronic devices. Here, kinetics of ET in trHb-Bs covalently attached via its surface amino acid residues either to COOH- or NH2-terminated (CH2)2-16 alkanethiol SAM assembled on gold are shown to depend on the alkanethiol length and functionalization, not being limited by electron tunnelling through the SAMs but gated by ET preceding reactions due to conformational changes in the heme active site/at the interface. ET gating was sensitive to the properties of SAMs that trHb-Bs interacted with.

Prevalence of Escherichia coli in surface waters of Southeast Asian cities.

Surface water samples were collected from rivers which fed into large urban areas within Vietnam, Indonesia, Cambodia, and Thailand and were processed to enumerate Escherichia coli. Selected isolates were further characterized using PCR to detect the presence of specific virulence genes. Analyzing the four countries together, the approximate mean cfu/100 ml for E.

High performance enzyme fuel cells using a genetically expressed FAD-dependent glucose dehydrogenase α-subunit of Burkholderia cepacia immobilized in a carbon nanotube electrode for low glucose conditions.

FAD-dependent glucose dehydrogenase (FAD-GDH) of Burkholderia cepacia was successfully expressed in Escherichia coli and subsequently purified in order to use it as an anode catalyst for enzyme fuel cells. The purified enzyme has a low Km value (high affinity) towards glucose, which is 463.8 μM, up to 2-fold exponential range lower compared to glucose oxidase.