The Biological Activities of Polyether Ionophore Antibiotic Routiennocin is Independent of Absolute Stereochemistry.

Carboxylic polyether ionophores (CPIs) are among the most prevalent agricultural antibiotics (notably in the US) and these compounds have been in use for decades. The potential to reposition CPIs beyond veterinary use, e. g. through chemical modifications to enhance their selectivity window, is an exciting challenge and opportunity, considering their general resilience towards resistance development. Given the very large societal impact of these somewhat controversial compounds, it is surprising that many aspects of their mechanisms and activities in cells remain unclear. Here, we report comparative biological activities of the CPI routiennocin and two stereoisomers, including its enantiomer. We used an efficient convergent synthesis strategy to access the compounds and conducted a broad survey of antibacterial activities against planktonic cells and biofilms as well as the compounds' effects on mammalian cells, the latter assessed both via standard cell viability assays and broad morphological profiling. Interestingly, similar bioactivity of the enantiomeric pair was observed across all assays, strongly suggesting that chiral interactions do not play a decisive role in the mode of action. Overall, our findings are consistent with a mechanistic model involving highly dynamic behaviour of CPIs in biological membranes.

A Self-Powered Biosensor for the Detection of Glutathione.

Glutathione is an important biological molecule which can be an indicator of numerous diseases. A method for self-powered detection of glutathione levels in solution has been developed using an enzymatic biofuel cell. The device consists of a glucose oxidase anode and a bilirubin oxidase cathode. For the detection of glutathione, the inhibition of bilirubin oxidase leads to a measurable decrease in current and power output. The reported method has a detection limit of 0.043 mM and a linear range up to 1.7 mM. Being able to detect a range of concentrations can be useful in evaluating a patient's health. This method has the potential to be implemented as a quick, low-cost alternative to previously reported methods.

Why Is Direct Glycosylation with N-Acetylglucosamine Donors Such a Poor Reaction and What Can Be Done about It?

The monosaccharide N-acetyl-d-glucosamine (GlcNAc) is an abundant building block in naturally occurring oligosaccharides, but its incorporation by chemical glycosylation is challenging since direct reactions are low yielding. This issue, generally agreed upon to be caused by an intermediate 1,2-oxazoline, is often bypassed by introducing extra synthetic steps to avoid the presence of the NHAc functional group during glycosylation. The present paper describes new fundamental mechanistic insights into the inherent challenges of performing direct glycosylation with GlcNAc. These results show that controlling the balance of oxazoline formation and glycosylation is key to achieving acceptable chemical yields. By applying this line of reasoning to direct glycosylation with a traditional thioglycoside donor of GlcNAc, which otherwise affords poor glycosylation yields, one may obtain useful glycosylation results.

Enzyme Immobilization and Mediation with Osmium Redox Polymers.

Enzymatic electrodes are becoming increasingly common for energy production and sensing applications. Research over the past several decades has addressed a major issue that can occur when using these biocatalysts, i.e., slow heterogeneous electron transfer, by incorporation of a redox active species to act as an electron shuttle. There are several advantages to immobilizing both the enzyme and mediator at the enzyme surface, including increased electron transfer rates, decreased enzyme leaching, and minimized diffusion limitations. Redox polymers consisting of a redox active center attached to a polymer backbone are a particularly attractive option because they have high self-exchange rates for electron transfer and tunable redox potential. Osmium (Os) polymers are the most well studied of this type of polymer for bioelectrocatalysis. Here, we describe the methods to synthesize one of the most common Os redox polymers and how it can be used to fabricate glucose oxidase electrodes. Procedures are also outlined for evaluating the enzymatic electrodes.

Enzymatic biofuel cells: 30 years of critical advancements.

Enzymatic biofuel cells are bioelectronic devices that utilize oxidoreductase enzymes to catalyze the conversion of chemical energy into electrical energy. This review details the advancements in the field of enzymatic biofuel cells over the last 30 years. These advancements include strategies for improving operational stability and electrochemical performance, as well as device fabrication for a variety of applications, including implantable biofuel cells and self-powered sensors. It also discusses the current scientific and engineering challenges in the field that will need to be addressed in the future for commercial viability of the technology.

Long-term arsenic monitoring with an Enterobacter cloacae microbial fuel cell.

A microbial fuel cell was constructed with biofilms of Enterobacter cloacae grown on the anode. Bioelectrocatalysis was observed when the biofilm was grown in media containing sucrose as the carbon source and methylene blue as the mediator. The presence of arsenic caused a decrease in bioelectrocatalytic current. Biofilm growth in the presence of arsenic resulted in lower power outputs whereas addition of arsenic showed no immediate result in power output due to the short term arsenic resistance of the bacteria and slow transport of arsenic across cellular membranes to metabolic enzymes. Calibration curves plotted from the maximum current and maximum power of power curves after growth show that this system is able to quantify both arsenate and arsenate with low detection limits (46 µM for arsenate and 4.4 µM for arsenite). This system could be implemented as a method for long-term monitoring of arsenic concentration in environments where arsenic contamination could occur and alter the metabolism of the organisms resulting in a decrease in power output of the self-powered sensor.

A protocol for metal triflate catalyzed direct glycosylations with GalNAc 1-OPiv donors.

Herein we report on the development of novel glycosylation methodology for the concise synthesis of naturally occurring glycoconjugate motifs containing N-acetylgalactosamine (GalNAc) from the cheaper and commercially available N-acetylglucosamine (GlcNAc). The stereoselective glycosylations proceed with catalytic amounts of a promoter and without the need for N-protection other than the biologically relevant N-acetyl group. Among the catalysts explored, both Bi(OTf)3 and Fe(OTf)3 were found to be highly active Lewis acids for this reaction. It was also found that other less reactive metal triflates such as those of Cu(II) and Yb(III) can be beneficial in glycosylation reactions on more demanding glycosyl acceptors. We have furthermore demonstrated that it is possible to control the anomeric stereoselectivity in the glycosylation via postglycosylation in situ anomerization to obtain good yields of α-galactosides. The present protocol was used to prepare important naturally occurring carbohydrate motifs, including a trisaccharide fragment of the naturally occurring marine sponge clarhamnoside.

Thylakoid direct photobioelectrocatalysis: utilizing stroma thylakoids to improve bio-solar cell performance.

Thylakoid membranes from spinach were separated into grana and stroma thylakoid fractions which were characterized by several methods (pigment content, protein gel electrophoresis, photosystem activities, and electron microscopy analysis) to confirm that the intact thylakoids were differentiated into the two domains. The results of photoelectrochemical experiments showed that stroma thylakoid electrodes generate photocurrents more than four times larger than grana thylakoids (51 ± 4 nA cm(-2) compared to 11 ± 1 nA cm(-2)). A similar trend was seen in a bio-solar cell configuration with stroma thylakoids giving almost twice the current (19 ± 3 µA cm(-2)) as grana thylakoids (11 ± 2 µA cm(-2)) with no change in open circuit voltage.

High performance thylakoid bio-solar cell using laccase enzymatic biocathodes.

Thylakoid membranes have previously been used for electrochemical solar energy conversion, but the current output and open circuit voltage are low, in part due to limitations of the cathode. In this paper, a thylakoid bioanode and laccase biocathode were combined in the construction of a bio-solar cell capable of light-induced generation of electrical power. This two-compartment cell showed a greater than 5-fold increase in short circuit current density and an open circuit voltage 0.275 V larger than that of a thylakoid bio-solar cell incorporating an air-breathing Pt cathode. The electrodes were then tested in several solutions of varying pH to evaluate the possibility of constructing a compartment-less bio-solar cell. This membrane-less cell, operating at pH 5.5, generated a short circuit photocurrent density of 14.0 ± 1.8 µA cm(-2) which is 25% larger than the two-compartment cell and a similar open circuit voltage of 0.720 ± 0.018 V.

An implantable biofuel cell for a live insect.

A biofuel cell incorporating a bienzymatic trehalase|glucose oxidase trehalose anode and a bilirubin oxidase dioxygen cathode using Os complexes grafted to a polymeric backbone as electron relays was designed and constructed. The specific power densities of the biofuel cell implanted in a female Blaberus discoidalis through incisions into its abdomen yielded maximum values of ca. 55 µW/cm(2) at 0.2 V that decreased by only ca. 5% after ca. 2.5 h of operation.

Bifunctional trehalose anode incorporating two covalently linked enzymes acting in series.

A bienzymatic electrode incorporating trehalase (Tre) and glucose oxidase (GOx) covalently bound to the surface of Pt through a functionalized thiol linker (Tre|GOx|Pt) has been designed and assembled and its catalytic properties examined by chemical and electrochemical methods in aqueous phosphate buffer solutions (PBS, pH 7.4). Exposure of Tre|GOx|Pt to PBS containing trehalose (Tr) and subsequent polarization at 0.6 V versus Ag/AgCl yielded after a few minutes well-defined steady-state currents ascribed to the oxidation of hydrogen peroxide generated by the GOx-mediated oxidation of glucose (Gl) produced by the Tre-mediated dissociation of Tr. Plots of the steady-state currents versus [Tr] over the range examined, i.e., 5-25 mM, were found to be linear. Implications of these results toward the development of an implantable biofuel cell as an autonomous energy conversion device for insects are discussed.