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PubMed2006

Making glucose oxidase fit for biofuel cell applications by directed protein evolution.

Progress in miniature chip-design raises demands for implantable power sources in health care applications such as continuous glucose monitoring of diabetic patients. Pioneered by Adam Heller, miniaturized enzymatic biofuel cells (mBCs) convert blood sugars into electrical energy by employing for example glucose oxidase (GOx) on the anode and bilirubin oxidase on the cathode. To match application demands it is crucial to increase lifetime and power output of mBCs. The power output has been limited by the performance of GOx on the anode. We developed a glucose oxidase detection assay (GODA) as medium-throughput screening system for improving GOx properties by directed protein evolution. GODA is a reaction product detection assay based on coupled enzymatic reactions leading to NADPH formation which is recorded at 340 nm. The main advantage of the assay is that it detects the production of d-gluconolactone instead of the side-product hydrogen peroxide and enables to improve bioelectrochemical properties of GOx. For validating the screening system, a mutagenic library of GOx from Aspergillus niger (EC 1.1.3.4) was generated and screened for improved activity using Saccharomyces cerevisiae as host. Directed evolution resulted in a GOx mutant I115V with 1.4-1.5-fold improved activity for beta-d-glucose (Vmax from 7.94 to 10.81 micromol min(-1) mg(-1); Km approximately 19-21 mM) and oxygen consumption kinetics correlate well [Vmax (O2) from 5.94 to 8.34 micromol min(-1) mg(-1); Km (O2) from 700 to 474 microM]. The developed mutagenic protocol and GODA represent a proof-of-principle that GOx can be evolved by directed evolution in S. cerevisiae for putative use in biofuel cells.

Bioelectric Energy Sources↗
PubMed2001

Emission factors of carbon monoxide and size-resolved aerosols from biofuel combustion.

This study reports emission factors of carbon monoxide and size-resolved aerosols from combustion of wood, dung cake, and biofuel briquette in traditional and improved stoves in India. Wood was the cleanest burning fuel, with higher emissions of CO from dung cake and particulate matter from both dung cake and briquette fuels. Combustion of dung cake, especially in an improved metal stove, resulted in extremely high pollutant emissions. Instead, biogas from anaerobic dung digestion should be promoted as a cooking fuel for public health protection. Pollutant emissions increased with increasing stove thermal efficiency, implying that thermal efficiency enhancement in the improved stoves was mainly from design features leading to increased heat transfer but not combustion efficiency. Compared to the traditional stove, the improved stoves resulted in the lower pollutant emissions on a kW h-1 basis from wood combustion but in similar emissions from briquette and dung cake. Stove designs are needed with good emissions performance across multiple fuels. Unimodal aerosol size distributions were measured from biofuel combustion with mass median aerodynamic diameters of 0.5-0.8 micron, about a factor of 10 larger than those from fossil fuel combustion (e.g. diesel), with potential implications for lung deposition and health risk.

Aerosols↗
PubMed2004

Radiocarbon apportionment of fossil versus biofuel combustion sources of polycyclic aromatic hydrocarbons in the Stockholm metropolitan area.

Source-diagnostic markers and the isotopic composition of polycyclic aromatic hydrocarbons (PAHs) were examined in surface sediments from the greater Stockholm waterways to deduce the contribution from biomass sources to the environmental PAH load. The summed concentration of 20 PAHs ranged from 0.8 to 45.1 microg/g (dry weight) and exhibited a steep decline with increasing distance from the city center evidencing that sources within the metropolitan area of Stockholm dominate its PAH burden. Several diagnostic PAH ratios indicated an overwhelming predominance of pyrogenic sources over the petrogenic ones, while retene and 1,7-dimethylphenanthrene were unable to correctly evaluate the contribution from biomass combustion. The stable carbon isotope composition (delta13C) of individual PAHs ranged from -24.8 to -27.0% but also was proved inefficient to discriminate between different types of fuels due to the overlapping signals in various sources. The delta14C values of PAHs ranged between -550.4 and -934.1%, indicating a clear predominance of fossil fuel sources. By using an isotopic mass balance approach, we estimated that on average 17+/-9% of PAHs derived from biomass combustion. This radiocarbon apportionment, in conjunction with detailed energy statistics for the Stockholm region, revealed that the ambient PAH burden is roughly similar, per unit energy produced, from fossil fuels and biofuels. Societies' shifting energy policies toward a larger reliance on biofuels may thus not lead to further deterioration of air quality and respiratory ailments for the urban population.

Bioelectric Energy Sources↗
PubMed2003

Characteristics of a miniature compartment-less glucose-O2 biofuel cell and its operation in a living plant.

We report the temperature, pH, glucose concentration, NaCl concentration, and operating atmosphere dependence of the power output of a compartment-less miniature glucose-O(2) biofuel cell, comprised only of two bioelectrocatalyst-coated carbon fibers, each of 7 micro m diameter and 2 cm length (Mano, N.; Mao, F.; Heller, A. J. Am. Chem. Soc. 2002, 124, 12962). The bioelectrocatalyst of the anode consists of glucose oxidase from Aspergillus niger electrically "wired" by polymer I, having a redox potential of -0.19 V vs Ag/AgCl. That of the cathode consists of bilirubin oxidase from Trachyderma tsunodae "wired" by polymer II having a redox potential of +0.36 V vs Ag/AgCl (Mano, N.; Kim, H.-H.; Zhang, Y.; Heller, A. J. Am. Chem. Soc. 2002, 124, 6480. Mano, N.; Kim, H.-H.; Heller, A. J. Phys. Chem. B 2002, 106, 8842). Implantation of the fibers in the grape leads to an operating biofuel cell producing 2.4 micro W at 0.52 V.

Bioelectric Energy Sources↗
PubMed2005

Magnetic field effects on bioelectrocatalytic reactions of surface-confined enzyme systems: enhanced performance of biofuel cells.

The effect of a constant magnetic field on bioelectrocatalytic transformations of three different enzyme assemblies linked to electrodes is examined and correlated with a theoretical magnetohydrodynamic model. The systems consist of surface-reconstituted glucose oxidase (GOx), an integrated lactate dehydrogenase/nicotinamide/pyrroloquinoline quinone assembly (LDH/NAD+ -PQQ), and a cytochrome c/cytochrome oxidase system (Cyt c/COx) linked to the electrodes. Pronounced effects of a constant magnetic field applied parallel to the electrode surface are observed for the bioelectrocatalyzed oxidation of glucose and lactate by the GOx-electrode and LDH/NAD+ -PQQ-electrode, respectively. The enhancement of the bioelectrocatalytic processes correlates nicely with the magnetohydrodynamic model, and the limiting current densities (iL) relate to B1/3 (B = magnetic flux density) and to C4/3 (C* = bulk concentration of the substrate). A small magnetic field effect is observed for the Cyt c/COx-electrode, and its origin is still questionable. The effect of the constant magnetic field on the performance of biofuel cells with different configurations is examined. For the biofuel cell consisting of LDH/NAD+ -PQQ anode and Cyt c/COx cathode, a 3-fold increase in the power output was observed at an applied magnetic field of B = 0.92 T and external load of 1.2 kOhms.

Bioelectric Energy Sources↗
PubMed2006

Soybean-derived biofuels and home heating fuels.

It is environmentally enticing to consider replacing or blending petroleum derived heating fuels with biofuels for many reasons. Major considerations include the soaring worldwide price of petroleum products, especially home heating oil, the toxicity of the petroleum-derived fuels and the environmental damage that leaking petroleum tanks afford. For these reasons, it has been suggested that domestic renewable energy sources be considered as replacements, or at the least, as blending stocks for home heating fuels. If recycled soy restaurant cooking oils could be employed for this purpose, this would represent an environmental advantage. Renewable plant sources of energy tend to be less toxic than their petroleum counterparts. This is an important consideration when tank leakage occurs. Home fuel oil storage tanks practically always contain some bottom water. This water environment has a pH value that factors into heating fuel stability. Therefore, the question is: would the biofuel help or exacerbate fuel stability and furnace maintenance issues?

Acids↗
PubMed2005

Residential biofuels in South Asia: carbonaceous aerosol emissions and climate impacts.

High concentrations of pollution particles, including "soot" or black carbon, exist over the Indian Ocean, but their sources and geographical origins are not well understood. We measured emissions from the combustion of biofuels, used widely in south Asia for cooking, and found that large amounts of carbonaceous aerosols are emitted per kilogram of fuel burnt. We calculate that biofuel combustion is the largest source of black carbon emissions in India, and we suggest that its control is central to climate change mitigation in the south Asian region.

Journal Article↗
PubMed2001

An electrochemical approach to the studies of biological redox reactions and their applications to biosensors, bioreactors, and biofuel cells.

Enzymatic redox reactions for the oxidation (or reduction) of the substrates use organic dyes or metal complexes as electron acceptors (or donors), which can be regenerated by electrochemical reactions of the compounds at an electrode surface. This type of coupling of the enzymatic reactions with the electrochemical reactions is called bioelectrocatalysis and allows us to measure the enzymatic reactions by an electrochemical method. The enzyme-electrochemical method provides a new technique for investigating a variety of biological redox reactions and for applying the reactions to biosensors, bioreactors, and biofuel cells. The bioelectrocatalysis-based research works are described here. First, a new method of protein redox potential measurements and a novel electrochemical kinetic analysis of oxidoreductase reactions are described. Second, a new methodology of characterizing microbial catalytic activities is presented. In the last, applications to the constructions of biosensors, bioreactors, and biofuel cells are mentioned.

Journal Article↗
PubMed2005

A low-cost biofuel cell with pH-dependent power output based on porous carbon as matrix.

A glucose/O2 biofuel cell (BFC) possessing a pH-dependent power output was fabricated by taking porous carbon (PC) as the matrix to load glucose oxidase or fungi laccase as the catalysts. The electrolytes in the anode and cathode compartments contain ferrocene monocarboxylic acid and 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt as the mediators, respectively. The power of the BFC was enhanced significantly by using PC as the matrix, rather than glassy carbon electrode. Additionally, the power output of the BFC decreases as the pH of the solution increases from 4.0 to 7.0, which provides a simple and efficient method to achieve the required power output. More importantly, the BFC can operate at pH 6.0, and even at pH 7.0, which overcomes the requirement for cathode solutions of pH<5.0 when using fungi laccase as a catalyst. Operation of the BFC at neutral pH may provide a means to power medical devices implanted in physiological systems. The facile and low-cost fabrication of this BFC may enable its development for other applications.

Carbon↗
PubMed1996

Measurement of free glycerol in biofuels.

A method has been developed to determine the free glycerol content in esters derived from vegetable oils. After extraction of glycerol, its measurement in small quantities in biofuels (methyl or ethyl esters) is possible by high-pressure liquid chromatography combined with pulsed amperometric detection, HPLC-PAD. The method has proved to be simple, rapid and accurate. The limit of detection is 1 microg/g.

Journal Article↗
PubMed2006

Alginate/carbon composite beads for laccase and glucose oxidase encapsulation: application in biofuel cell technology.

Alginate-carbon beads were prepared in order to develop a biocompatible matrix for laccase and glucose oxidase immobilization for application in biofuel cell technology. The enzyme loading capacity was high (91%) in pure alginate beads for glucose oxidase. For laccase, the loading capacity was enhanced from 75% to 83% by introducing carbon. Desorption out of the matrix was controlled by the enzymes' diffusion and reached a plateau after 40 h for laccase and 70 h for glucose oxidase. Two-thirds of both enzymes was irreversibly retained inside the alginate beads. This proportion increased to 80% for laccase in combined alginate/carbon beads. Half-life of the adsorbed enzyme was enhanced to 74 days for laccase in carbon/alginate beads and 45 days for glucose oxidase in pure alginate as compared to 38 days and 23 days for free enzymes, respectively.

Adsorption↗
PubMed2007

Catalytic cracking of palm oil for the production of biofuels: optimization studies.

Oil palm is widely grown in Malaysia. Palm oil has attracted the attention of researchers to develop an 'environmentally friendly' and high quality fuel, free of nitrogen and sulfur. In the present study, the catalytic cracking of palm oil to biofuel was studied over REY catalyst in a transport riser reactor at atmospheric pressure. The effect of reaction temperature (400-500 degrees C), catalyst/palm oil ratio (5-10) and residence time (10-30s) was studied over the yield of bio-gasoline and gas as fuel. Design of experiments was used to study the effect of operating variables over conversion of palm oil and yield of hydrocarbon fuel. The response surface methodology was used to determine the optimum value of the operating variables for maximum yield of bio-gasoline fraction in the liquid product obtained.

Catalysis↗
PubMed2006

Plant genetic engineering to improve biomass characteristics for biofuels.

Currently, most ethanol produced in the United States is derived from maize kernel, at levels in excess of four billion gallons per year. Plant lignocellulosic biomass is renewable, cheap and globally available at 10-50 billion tons per year. At present, plant biomass is converted to fermentable sugars for the production of biofuels using pretreatment processes that disrupt the lignocellulose and remove the lignin, thus allowing the access of microbial enzymes for cellulose deconstruction. Both the pretreatments and the production of enzymes in microbial tanks are expensive. Recent advances in plant genetic engineering could reduce biomass conversion costs by developing crop varieties with less lignin, crops that self-produce cellulase enzymes for cellulose degradation and ligninase enzymes for lignin degradation, or plants that have increased cellulose or an overall biomass yield.

Biomass↗
PubMedDate not supplied

Investigation on organic pollutants from a domestic heating system using various solid biofuels.

Various herbaceous biofuels (straw, whole plant cereals and set aside hay) and spruce wood were tested for their potential to form PCDD/F, PCPh, PCBz and PAH during combustion. The trials were conducted in an automatically charged multi-fuel furnace for domestic applications (50 kWth). Both, flue gas and the different ash fractions were analysed. CO-emission results show, that combustion conditions were relatively uniform (mean CO-level: 200 mg/m3 at 13% O2 in flue gas). Likewise, the TOC- and PAH-emissions in the fuel gas remained constantly on a relatively low level. However, for the PCDD/F, PCPh and PCBz increased emissions were detected when herbacious fuels were applied. This may be attributed to their higher chlorine concentration and the high ash content, which is responsible for increased dust emissions during combustion. Similar observations were also found for the PCDD/F-concentrations in the ashes. Combustion chamber ashes usually showed a drastically reduced contamination with highly toxical compounds, compared to the ash fraction from the heat exchanger ash or to chimney soot.

Air Pollutants↗
PubMed2002

The yield and composition of switchgrass and coastal panic grass grown as a biofuel in southern England.

Switchgrass (Panicum virgatum L.) and coastal panic grass (Panicum amarum A.S. Hitchc. & Chase) are perennial grasses indigenous to North America. Switchgrass has been shown to have good potential as a biofuel crop in both the US and Canada. In the study reported here, seven varieties of switchgrass and one panic grass were evaluated for 5 years under the temperate maritime conditions in Southern England. Both species had 0 or 60 kg N ha(-1) applied annually in spring as treatment. Yield was measured after flowering and when stems were dead in the winter. Yield increased annually for 4-5 years except for the variety Dacotah, and in the fifth year dead stem yields ranged from 8.82 to 13.97 t dm ha(-1). There was no response to N except for one variety in one year. Mineral concentration in biomass was higher at flowering than at dead stem harvest and delaying harvesting further provided more time for P, K and Cl to be leached but yield also declined.

Agriculture↗
PubMed2002

Immobilisation of enzymes on poly(aniline)-poly(anion) composite films. Preparation of bioanodes for biofuel cell applications.

Immobilisation of enzymes is important for applications such as biosensors or biofuel cells. A poly(histidine) tag had been introduced on the C terminus of a lactate dehydrogenase enzyme. This mutant enzyme was then immobilised onto poly(aniline) (PANi)-poly(anion) composite films, PANi-poly(vinylsulfonate) (PVS) or PANi-poly(acrylate) (PAA). The NADH produced by the immobilised enzyme in the presence of beta-nicotinamide adenine dinucleotide (NAD(+)) and lactate is oxidised at the poly(aniline)-coated electrode at 0.05 to 0.1 V vs. saturated calomel electrode (SCE) at 35 degrees C.

Aniline Compounds↗
PubMed2003

Bioelectrocatalysis-based application of quinoproteins and quinoprotein-containing bacterial cells in biosensors and biofuel cells.

Electrochemical studies on the applied aspects of quinoproteins are briefly reviewed. Catalytic reactions of quinoprotein enzymes can be connected to electrochemical reactions directly or by the mediation of molecules functioning as electron acceptors of the enzymes. Such an enzyme-electrochemical reaction is called bioelectrocatalysis. It provides a novel method of kinetic analysis of enzyme catalysis and even whole bacterial cell catalysis. The principle of bioelectrocatalysis is first described, then, the bioelectrocatalysis-based application of quinoproteins in biosensors is mentioned. Characteristics and performance of this type of biosensor is explained by citing our own work. Possible application in bioreactors and biofuel cells is also mentioned.

Bacteria↗