Highly stable enzyme precipitate coatings and their electrochemical applications

This paper describes highly stable enzyme precipitate coatings (EPCs) on electrospun polymer nanofibers and carbon nanotubes (CNTs), and their potential applications in the development of highly sensitive biosensors and high-powered biofuel cells. EPCs of glucose oxidase (GOx) were prepared by precipitating GOx molecules in the presence of ammonium sulfate, then cross-linking the precipitated GOx aggregates on covalently attached enzyme molecules on the surface of nanomaterials. EPCs-GOx not only improved enzyme loading, but also retained high enzyme stability. For example, EPC-GOx on CNTs showed a 50 times higher activity per unit weight of CNTs than the conventional approach of covalent attachment, and its initial activity was maintained with negligible loss for 200 days. EPC-GOx on CNTs was entrapped by Nafion to prepare enzyme electrodes for glucose sensors and biofuel cells. The EPC-GOx electrode showed a higher sensitivity and a lower detection limit than an electrode prepared with covalently attached GOx (CA-GOx). The CA-GOx electrode showed an 80% drop in sensitivity after thermal treatment at 50°C for 4 h, while the EPC-GOx electrode maintained its high sensitivity with negligible decrease under the same conditions. The use of EPC-GOx as the anode of a biofuel cell improved the power density, which was also stable even after thermal treatment of the enzyme anode at 50°C. The excellent stability of the EPC-GOx electrode together with its high current output create new potential for the practical applications of enzyme-based glucose sensors and biofuel cells.     

Citric acid cycle biomimic on a carbon electrode

The citric acid cycle is one of the main metabolic pathways living cells utilize to completely oxidize biofuels to carbon dioxide and water. The overall goal of this research is to mimic the citric acid cycle at the carbon surface of an electrode in order to achieve complete oxidation of ethanol at a bioanode to increase biofuel cell energy density. In order to mimic this process, dehydrogenase enzymes (known to be the electron or energy producing enzymes of the citric acid cycle) are immobilized in cascades at an electrode surface along with non-energy producing enzymes necessary for the cycle to progress. Six enzymatic schemes were investigated each containing an additional dehydrogenase enzyme involved in the complete oxidation of ethanol. An increase in current density is observed along with an increase in power density with each additional dehydrogenase immobilized on an electrode, reflecting increased electron production at the bioanode with deeper oxidation of the ethanol biofuel. By mimicking the complete citric acid cycle on a carbon electrode, power density was increased 8.71-fold compared to a single enzyme (alcohol dehydrogenase)-based ethanol/air biofuel cell.     

Enzyme precipitate coatings of glucose oxidase onto carbon paper for biofuel cell applications

Enzymatic biofuel cells (BFC) have a great potential as a small power source, but their practical applications are being hampered by short lifetime and low power density. This study describes the direct immobilization of glucose oxidase (GOx) onto the carbon paper in the form of highly stable and active enzyme precipitation coatings (EPCs), which can improve the lifetime and power density of BFCs. EPCs were fabricated directly onto the carbon paper via a three-step process: covalent attachment (CA), enzyme precipitation, and chemical crosslinking. GOx-immobilized carbon papers via the CA and EPC approaches were used as an enzyme anode and their electrochemical activities were tested under the BFC-operating mode. The BFCs with CA and EPC enzyme anodes produced the maximum power densities of 50 and 250 μW/cm(2) , respectively. The BFC with the EPC enzyme anode showed a stable current density output of >700 μA/cm(2) at 0.18 V under continuous operation for over 45 h. When a maple syrup was used as a fuel under ambient conditions, it also produced a stable current density of >10 μA/cm(2) at 0.18 V for over 25 h. It is anticipated that the direct immobilization of EPC on hierarchical-structured electrodes with a large surface area would further improve the power density of BFCs that can make their applications more feasible.     

Development of Dopant‐Free Organic Hole Transporting Materials for Perovskite Solar Cells

There has been considerable progress over the last decade in development of the perovskite solar cells (PSCs), with reported performances now surpassing 25.2% power conversion efficiency. Both long‐term stability and component costs of PSCs remain to be addressed by the research community, using hole transporting materials (HTMs) such as 2,2′,7,7′‐tetrakis(N,N′‐di‐pmethoxyphenylamino)‐9,9′‐spirbiuorene(Spiro‐OMeTAD) and poly[bis(4‐phenyl)(2,4,6‐trimethylphenyl)amine] (PTAA). HTMs are essential for high‐performance PSC devices. Although effective, these materials require a relatively high degree of doping with additives to improve charge mobility and interlayer/substrate compatibility, introducing doping‐induced stability issues with these HTMs, and further, additional costs and experimental complexity associated with using these doped materials. This article reviews dopant‐free organic HTMs for PSCs, outlining reports of structures with promising properties toward achieving low‐cost, effective, and scalable materials for devices with long‐term stability. It summarizes recent literature reports on non‐doped, alternative, and more stable HTMs used in PSCs as essential components for high‐efficiency cells, categorizing HTMs as reported for different PSC architectures in addition to use of dopant‐free small molecular and polymeric HTMs. Finally, an outlook and critical assessment of dopant‐free organic HTMs toward commercial application and insight into the development of stable PSC devices is provided.     

Development of a novel glucose enzyme fuel cell system employing protein engineered PQQ glucose dehydrogenase

Glucose dehydrogenase harboring pyrroloquinoline quinone as the prosthetic group (PQQGDH) from Acinetobacter calcoaceticus is an ideal enzyme for the anode of biofuel cell, because of its oxygen insensitivity and high catalytic efficiency. However, the application of PQQGDH for the bioanode is inherently limited because of its instability. Using Ser415Cys mutant whose stability was greatly improved, we constructed the biofuel cell system employing the engineered PQQGDH as the bioanode enzyme and bilirubin oxidase (BOD) as the biocathode, and compared the stability of the biofuel cell with that employing wild-type PQQGDH. The maximum power density was 17.6 microW/cm2 at an external optimal load of 200 k omega. Using Ser415Cys mutant, the lifetime of the biofuel cell system was greatly extended to 152 h, more than six times as that of the biofuel cell employing the wild-type.     

Enzyme-based biofuel cells

Enzyme-based biofuel cells possess several positive attributes for energy conversion, including renewable catalysts, flexibility of fuels (including renewables), and the ability to operate at room temperature. However, enzyme-based biofuel cells remain limited by short lifetimes, low power densities and inefficient oxidation of fuels. Recent advances in biofuel cell technology have addressed these deficiencies and include methods to increase lifetime and environmental stability.     

Application of metagenomic techniques in mining enzymes from microbial communities for biofuel synthesis

Feedstock for biofuel synthesis is transitioning to lignocelluosic biomass to address criticism over competition between first generation biofuels and food production. As microbial catalysis is increasingly applied for the conversion of biomass to biofuels, increased import has been placed on the development of novel enzymes. With revolutionary advances in sequencer technology and metagenomic sequencing, mining enzymes from microbial communities for biofuel synthesis is becoming more and more practical. The present article highlights the latest research progress on the special characteristics of metagenomic sequencing, which has been a powerful tool for new enzyme discovery and gene functional analysis in the biomass energy field. Critical enzymes recently developed for the pretreatment and conversion of lignocellulosic materials are evaluated with respect to their activity and stability, with additional explorations into xylanase, laccase, amylase, chitinase, and lipolytic biocatalysts for other biomass feedstocks.     

A mediated glucose/oxygen enzymatic fuel cell based on printed carbon inks containing aldose dehydrogenase and laccase as anode and cathode

Enzyme electrodes show great potential for many applications, as biosensors and more recently as anodes and cathodes in biocatalytic fuel cells for power generation. Enzymes have advantages over metal catalysts, as they provide high specificity and reaction rates, while operating under mild conditions. Here we report on studies related to development of mass-producible, completely enzymatic printed glucose/oxygen biofuel cells. The cells are based on filter paper coated with conducting carbon inks containing mediators and laccase, for reduction of oxygen, or aldose dehydrogenase, for oxidation of glucose. Mediator performance in these printed formats is compared to relative rate constants for the enzyme-mediator reaction in solution, for a range of anode and cathode mediators. The power output and stability of fuels cells using an acidophilic laccase isolated from Trametes hirsuta is greater, at pH 5, than that for cells based on Melanocarpus albomyces laccase, that shows optimal activity closer to neutral pH, at pH 6. Highest power output, although of limited stability, was observed for ThL/ABTS cathodes, providing a maximum power density of 3.5 μWcm(-2) at 0.34 V, when coupled to an ALDH glucose anode mediated by an osmium complex. The stability of cell voltage above a threshold of 200 mV under a moderate 75 kΩ load is used to benchmark printed fuel cell performance. Highest stability was obtained for a printed fuel cell using osmium complexes as mediators of glucose oxidation by aldose dehydrogenase, and oxygen reduction by T. hirsuta laccase, maintaining cell voltage above 200 mV for 137 h at pH 5. These results provide promising directions for further development of mass-producible, completely enzymatic, printed biofuel cells.     

Recent trends in nanomaterials immobilised enzymes for biofuel production

Application of nanomaterials as novel supporting materials for enzyme immobilisation has generated incredible interest in the biotechnology community. These robust nanostructured forms, such as nanoparticles, nanofibres, nanotubes, nanoporous, nanosheets, and nanocomposites, possess a high surface area to volume ratios that can cause a high enzyme loading and facilitate reaction kinetics, thus improving biocatalytic efficiency for industrial applications. In this article, we discuss research opportunities of nanoscale materials in enzyme biotechnology and highlight recent developments in biofuel production using advanced material supports for enzyme immobilisation and stabilisation. Synthesis and functionalisation of nanomaterial forms using different methods are highlighted. Various simple and effective strategies designed to result in a stable, as well as functional protein-nanomaterial conjugates are also discussed. Analytical techniques confirming enzyme loading on nanomaterials and assessing post-immobilisation changes are discussed. The current status of versatile nanomaterial support for biofuel production employing cellulases and lipases is described in details. This report concludes with a discussion on the likely outcome that nanomaterials will become an integral part of sustainable bioenergy production.     

An ultrasonically enhanced inclined settler for microalgae harvesting

Microalgae have vast potential as a sustainable and scalable source of biofuels and bioproducts. However, algae dewatering is a critical challenge that must be addressed. Ultrasonic settling has already been exploited for concentrating various biological cells at relatively small batch volumes and/or low throughput. Typically, these designs are operated in batch or semicontinuous mode, wherein the flow is interrupted and the cells are subsequently harvested. These batch techniques are not well suited for scaleup to the throughput levels required for harvesting microalgae from the large‐scale cultivation operations necessary for a viable algal biofuel industry. This article introduces a novel device for the acoustic harvesting of microalgae. The design is based on the coupling of the acoustophoretic force, acoustic transparent materials, and inclined settling. A filtration efficiency of 70 ± 5% and a concentration factor of 11.6 ± 2.2 were achieved at a flow rate of 25 mL·min^?1 and an energy consumption of 3.6 ± 0.9 kWh·m^?3. The effects of the applied power, flow rate, inlet cell concentration, and inclination were explored. It was found that the filtration efficiency of the device is proportional to the power applied. However, the filtration efficiency experienced a plateau at 100 W L^?1 of power density applied. The filtration efficiency also increased with increasing inlet cell concentration and was inversely proportional to the flow rate. It was also found that the optimum settling angle for maximum concentration factor occurred at an angle of 50 ± 5°. At these optimum conditions, the device had higher filtration efficiency in comparison to other similar devices reported in the previous literature. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 31:414–423, 2015     

CO2生物转化关键酶固定体系构筑研究进展与挑战

酶催化CO₂还原制备高值化学品对缓解全球环境和能源危机具有重要意义,利用甲酸脱氢酶(formate dehydrogenase,FDH)或多酶级联还原CO₂制备甲酸/甲醇具有选择性高、条件温和的优势,但关键酶活性低、稳定性差和重复利用率低的问题限制了其规模化应用,酶的固定化为这些问题提供了有效解决方案。本文总结了近年来利用膜、无机材料、金属有机框架和共价有机框架等载体对酶进行固定化的研究进展,阐释了不同固定材料和固定方式的特点和优势;进一步总结了固定化酶与电催化或光催化耦联反应体系对CO₂还原的协同效果及应用,同时指出酶固定化技术和耦联反应体系目前存在的问题并对其发展前景进行了展望。     

Recent Advances in Design and Fabrication of Electrochemical Supercapacitors with High Energy Densities

In recent years, tremendous research effort has been aimed at increasing the energy density of supercapacitors without sacrificing high power capability so that they reach the levels achieved in batteries and at lowering fabrication costs. For this purpose, two important problems have to be solved: first, it is critical to develop ways to design high performance electrode materials for supercapacitors; second, it is necessary to achieve controllably assembled supercapacitor types (such as symmetric capacitors including double‐layer and pseudo‐capacitors, asymmetric capacitors, and Li‐ion capacitors). The explosive growth of research in this field makes this review timely. Recent progress in the research and development of high performance electrode materials and high‐energy supercapacitors is summarized. Several key issues for improving the energy densities of supercapacitors and some mutual relationships among various effecting parameters are reviewed, and challenges and perspectives in this exciting field are also discussed. This provides fundamental insight into supercapacitors and offers an important guideline for future design of advanced next‐generation supercapacitors for industrial and consumer applications.     

Improving the efficiency of enzyme utilization for sugar beet pulp hydrolysis

Sugar beet pulp (SBP) is a carbohydrate-rich residue of table sugar processing. It shows promise as a feedstock for fermentable sugar and biofuel production via enzymatic hydrolysis and microbial fermentation. This research focused on the enzymatic hydrolysis of SBP and examined the effects of solid loading (2–10?%, dry basis), enzyme preparation, and enzyme recycle on the production of fermentable sugars. The enzyme partitioning to the solid and liquid phases during SBP enzymatic hydrolysis and loss during recycling were investigated using SDS-PAGE and Zymogram analysis. Without considering product inhibition, the cellulase added initially to the SBP hydrolysis lost only 6?% filter paper activity and negligible carboxymethyl cellulose activity upon multiple cycles of SBP hydrolysis. It was found that enzyme dosage can be reduced by 50?% while maintaining similar, and in some cases higher fermentable sugar yield. The removal of hydrolysis products will further improve enzymatic hydrolysis of SBP for biofuel production.     

Barrier Designs in Perovskite Solar Cells for Long‐Term Stability

Perovskite solar cells (PSCs) have attracted much attention in the past decade and their power conversion efficiency has been rapidly increasing to 25.2%, which is comparable with commercialized solar cells. Currently, the long‐term stability of PSCs remains as a major bottleneck impeding their future commercial applications. Beyond strengthening the perovskite layer itself and developing robust external device encapsulation/packaging technology, integration of effective barriers into PSCs has been recognized to be of equal importance to improve the whole device’s long‐term stability. These barriers can not only shield the critical perovskite layer and other functional layers from external detrimental factors such as heat, light, and H₂O/O₂, but also prevent the undesired ion/molecular diffusion/volatilization from perovskite. In addition, some delicate barrier designs can simultaneously improve the efficiency and stability. In this review article, the research progress on barrier designs in PSCs for improving their long‐term stability is reviewed in terms of the barrier functions, locations in PSCs, and material characteristics. Regarding specific barriers, their preparation methods, chemical/photoelectronic/mechanical properties, and their role in device stability, are further discussed. On the basis of these accumulative efforts, predictions for the further development of effective barriers in PSCs are provided at the end of this review.     

Metabolic engineering for advanced biofuels production from Escherichia coli

Global energy and environmental problems have stimulated increasing efforts toward synthesizing liquid biofuels as transportation energy. Compared to the traditional biofuel, ethanol, advanced biofuels should offer advantages such as higher energy density, lower hygroscopicity, lower vapor pressure, and compatibility with existing transportation infrastructure. However, these fuels are not synthesized economically using native organisms. Metabolic engineering offers an alternative approach in which synthetic pathways are engineered into user-friendly hosts for the production of these fuel molecules. These hosts could be readily manipulated to improve the production efficiency. This review summarizes recent progress in the engineering of Escherichia coli to produce advanced biofuels.     

纳米材料固定化酶的研究进展

近年来,纳米技术为酶固定化提供了多种纳米级材料,纳米材料固定化酶不仅具有高的酶负载量,而且具有良好的酶稳定性。本文基于纳米材料固定化酶,对纳米材料的种类进行了总结,分析了纳米材料对固定化酶性能的影响,并介绍了纳米级固定化方法及纳米材料固定化酶在生物转化、生物传感器、生物燃料电池等领域的应用。     

Nanobiocatalysis and its potential applications

Nanobiocatalysis, in which enzymes are incorporated into nanostructured materials, has emerged as a rapidly growing area. Nanostructures, including nanoporous media, nanofibers, carbon nanotubes and nanoparticles, have manifested great efficiency in the manipulation of the nanoscale environment of the enzyme and thus promise exciting advances in many areas of enzyme technology. This review will describe these recent developments in nanobiocatalysis and their potential applications in various fields, such as trypsin digestion in proteomic analysis, antifouling, and biofuel cells.     

Enzymatic hydrolysis of cellulose coupled with electricity generation in a microbial fuel cell

Electricity can be directly generated by bacteria in microbial fuel cells (MFCs) from a variety of biodegradable substrates, including cellulose. Particulate materials have not been extensively examined for power generation in MFCs, but in general power densities are lower than those produced with soluble substrates under similar conditions likely as a result of slow hydrolysis rates of the particles. Cellulases are used to achieve rapid conversion of cellulose to sugar for ethanol production, but these enzymes have not been previously tested for their effectiveness in MFCs. It was not known if cellulases would remain active in an MFC in the presence of exoelectrogenic bacteria or if enzymes might hinder power production by adversely affecting the bacteria. Electricity generation from cellulose was therefore examined in two-chamber MFCs in the presence and absence of cellulases. The maximum power density with enzymes and cellulose was 100 +/- 7 mW/m(2) (0.6 +/- 0.04 W/m(3)), compared to only 12 +/- 0.6 mW/m(2) (0.06 +/- 0.003 W/m(3)) in the absence of the enzymes. This power density was comparable to that achieved in the same system using glucose (102 +/- 7 mW/m(2), 0.56 +/- 0.038 W/m(3)) suggesting that the enzyme successfully hydrolyzed cellulose and did not otherwise inhibit electricity production by the bacteria. The addition of the enzyme doubled the Coulombic efficiency (CE) to CE = 51% and increased COD removal to 73%, likely as a result of rapid hydrolysis of cellulose in the reactor and biodegradation of the enzyme. These results demonstrate that cellulases do not adversely affect exoelectrogenic bacteria that produce power in an MFC, and that the use of these enzymes can increase power densities and reactor performance.     

Theoretical Investigation of 2D Layered Materials as Protective Films for Lithium and Sodium Metal Anodes

Rechargeable batteries based on lithium (sodium) metal anodes have been attracting increasing attention due to their high capacity and energy density, but the implementation of lithium (sodium) metal anode still faces many challenges, such as low Coulombic efficiency and dendrites growth. Layered materials have been used experimentally as protective films (PFs) to address these issues. In this work, the authors explore using first‐principles computations the key factors that determine the properties and feasibility of various 2D layered PFs, including the defect pattern, crystalline structure, bond length, and metal proximity effect, and perform the simulations on both aspects of Li⁺ (Na⁺) ion diffusion property and mechanical stability. It is found that the introduction of defect, the increase in bond length, and the proximity effect by metal can accelerate the transfer of Li⁺ (Na⁺) ion and improve the ionic conductivity, but all of them make negative influences on the stiffness of materials against the suppression of dendrite growth and weaken both critical strains and critical stress. The results provide new insight into the interaction mechanism between Li⁺ (Na⁺) ions and PF materials at the atomic level and shed light onto exploring a variety of layered PF materials in metal anode battery systems.     

Development of nanostructured bioanodes containing dendrimers and dehydrogenases enzymes for application in ethanol biofuel cells

This paper describes the use of the electrostatic layer-by-layer (LbL) technique for the preparation of bioanodes with potential application in ethanol/O(2) biofuel cells. More specifically, the LbL technique was employed for immobilization of dehydrogenase enzymes and polyamidoamine (PAMAM) dendrimers onto carbon paper support. Both mono (anchoring only the enzyme alcohol dehydrogenase, ADH) and bi-enzymatic (anchoring both ADH and aldehyde dehydrogenase, AldDH) systems were tested. The amount of ADH deposited onto the Toray? paper was 95 ng cm(-2) per bilayer. Kinetic studies revealed that the LbL technique enables better control of enzyme disposition on the bioanode, as compared with the results obtained with the bioanodes prepared by the passive adsorption technique. The power density values achieved for the mono-enzymatic system as a function of the enzyme load ranged from 0.02 to 0.063 mW cm(-2) for the bioanode containing 36 ADH bilayers. The bioanodes containing a gas diffusion layer (GDL) displayed enhanced performance, but their mechanical stability must be improved. The bi-enzymatic system generated a power density of 0.12 mW cm(-2). In conclusion, the LbL technique is a very attractive approach for enzyme immobilization onto carbon platform, since it enables strict control of enzyme disposition on the bioanode surface with very low enzyme consumption.