Influence of artificial mediators on yeast-based fuel cell performance

Soluble artificial mediators are often applied to enhance the electron transfer from living cells to an anode in microbial fuel cells. Recently, we have demonstrated that the Candida melibiosica 2491 yeast strain possesses electrogenic properties and can be used as a biocatalyst in yeast-based fuel cells even in the absence of artificial mediators. To enhance the generated electrical power, the potential application of several organic compounds as mediators in a C. melibiosica-based fuel cell was examined in this study. The choice of compounds was based upon observed cyclic voltammetry reversible electrochemical behavior at potentials appropriate for mediated electron transfer. Among the studied mediators, methylene blue, methyl orange, methyl red and neutral red significantly increased the current and power outputs in comparison with those obtained with a mediatorless yeast-based fuel cell.     

Affinity of microbial fuel cell biofilm for the anodic potential

In analogy to the well established dependency of microbial reactions on the redox potential of the terminal electron acceptor, the dependency of the microbial activity in a highly active microbial fuel cell on the potential of the electron-accepting electrode (anode) in a microbial fuel cell (MFC) is investigated. An acetate-fed, pH-controlled MFC was operated for over 200 days to establish a highly active MFC anodic biofilm using ferricyanide as the catholyte and granular graphite as electrode material. From the Coulombic efficiency of 83% of the MFC the microbial activity could be recorded by online monitoring of the current. Our results suggest that (1) in analogy to the Michaelis-Menten kinetics a half-saturation anodic potential (here termed k(AP) value) could be established at which the microbial metabolic rate reached half its maximum rate. This k(AP) value was about -455 mV (vs Ag/AgCl) for our acetate-driven MFC and independent of the oxidation capacity of the cathodic half-cell; (2) a critical AP (here termed AP(crit)) of about -420 mV (vs Ag/AgCl) was established that characterizes the bacterial saturation by the electron-accepting system. This critical potential appeared to characterize the maximum power output of the MFC. This information would be useful for modeling and optimization of microbial fuel cells and the relative comparison of different microbial consortia at the anode.     

Microbial fuel cell using anaerobic respiration as an anodic reaction and biomineralized manganese as a cathodic reactant

We have operated a microbial fuel cell in which glucose was oxidized by Klebsiella pneumoniae in the anodic compartment, and biomineralized manganese oxides, deposited by Leptothrix discophora, were electrochemically reduced in the cathodic compartment. In the anodic compartment, to facilitate the electron transfer from glucose to the graphite electrode, we added a redox mediator, 2-hydroxy-1,4-naphthoquinone. We did not add any redox mediator to the cathodic compartment because the biomineralized manganese oxides were deposited on the surface of a graphite electrode and were reduced directly by electrons from the electrode. We have demonstrated that biomineralized manganese oxides are superiorto oxygen when used as cathodic reactants in microbial fuel cells. The current density delivered by using biomineralized manganese oxides as the cathodic reactant was almost 2 orders of magnitude higher than that delivered using oxygen. Several fuel cells were operated for 500 h, reaching anodic potentials of -441.5 +/- 31 mVscE and cathodic potentials of +384.5 +/- 64 mVscE. When the electrodes were connected by a 50 Ohms resistor, the fuel cell delivered the peak power density of 126.7 +/- 31.5 mW/m2.     

The role of soil microbes in the global carbon cycle: tracking the below‐ground microbial processing of plant‐derived carbon for manipulating carbon dynamics in agricultural systems

It is well known that atmospheric concentrations of carbon dioxide (CO₂) (and other greenhouse gases) have increased markedly as a result of human activity since the industrial revolution. It is perhaps less appreciated that natural and managed soils are an important source and sink for atmospheric CO₂ and that, primarily as a result of the activities of soil microorganisms, there is a soil‐derived respiratory flux of CO₂ to the atmosphere that overshadows by tenfold the annual CO₂ flux from fossil fuel emissions. Therefore small changes in the soil carbon cycle could have large impacts on atmospheric CO₂ concentrations. Here we discuss the role of soil microbes in the global carbon cycle and review the main methods that have been used to identify the microorganisms responsible for the processing of plant photosynthetic carbon inputs to soil. We discuss whether application of these techniques can provide the information required to underpin the management of agro‐ecosystems for carbon sequestration and increased agricultural sustainability. We conclude that, although crucial in enabling the identification of plant‐derived carbon‐utilising microbes, current technologies lack the high‐throughput ability to quantitatively apportion carbon use by phylogentic groups and its use efficiency and destination within the microbial metabolome. It is this information that is required to inform rational manipulation of the plant–soil system to favour organisms or physiologies most important for promoting soil carbon storage in agricultural soil. © 2014 The Authors. Journal of the Science of Food and Agriculture published by JohnWiley & Sons Ltd on behalf of Society of Chemical Industry.     

Comparative analysis of the fermentation performance of high‐quality milk beer strains (Kluyveromyces marxianus) and optimisation of medium formula for high‐density fermentation

Milk beer is a new type of dairy beverage in China and increasingly popular with consumers. The strain FXJ1 isolated in our laboratory was an excellent strain for the production of milk beer with alcohol content of 0.48%. Our research verified that the strain FXJ1 was superior to the commercial strain MJ for milk beer production and got the optimal medium formula (67.37 g/L sucrose, 29.7 g/L yeast extract, 15.61 g/L corn steep liquor, 4.13 g/L KH₂PO₄ and 0.3 g/L MgSO₄) which allowed the biomass production of strain FXJ1 to achieve 8.88 ± 0.08 g/L and increase 2.94‐fold as compared with yeast extract peptone dextrose base medium.     

Efficient very‐high‐gravity fermentation of sugarcane molasses by industrial yeast strains

In this study industrial strains were inoculated, in successive cycles, at high cell density into a sugarcane‐based juice containing normal (22%, w/v) and very high sucrose (30%, w/v) levels and supplemented with peptone as a nitrogen source. At 30 °C, in shaken cultures, in the normal gravity situation, efficient sucrose utilization occurred in both the supplemented and unsupplemented medium. At higher sugar concentrations, supplementation with peptone induced a more efficient fermentation compared with the unsupplemented medium, with higher biomass accumulation and maintenance of cell viability, and ethanol levels as high as 16% (v/v). Trehalose was also high during the cycles, probably as a necessary response of the yeast to the high stress fermentation conditions. This suggests that it is possible to increase ethanol production by using very‐high‐fermentation technology and that nutritional supplementation would have a positive effect on the fermentation, allowing for efficient sugar consumption and cell viability maintenance. Copyright © 2016 The Institute of Brewing & Distilling     

Isolation of obligate anaerobic rumen bacteria capable of degrading the neurotoxin β‐ODAP (β‐N‐oxalyl‐L‐α,β‐diaminopropionic acid) as evaluated by a liquid chromatography/biosensor analysis system

Six pure strains of obligate anaerobes capable of degrading the toxin β‐N‐oxalyl‐L ‐α, β‐diaminopropionic acid (β‐ODAP) contained in grass pea (Lathyrus sativus) have been isolated from cow rumen. The new isolates were identified as Megasphaera elsdenii (five different genotypes) and Clostridium bifermentans using 16S rDNA analysis. The β‐ODAP degrading efficiency of the isolates was evaluated by measuring the amount of β‐ODAP in the growth medium, which contained β‐ODAP as the only carbon source, before and after incubation with the microbes. The method of analysis was liquid chromatography employing bioelectrochemical detection. The biosensor is based on co‐immobilising two enzymes, glutamate oxidase (GlOx) and horseradish peroxidase (HRP), on the end of a spectrographic graphite electrode. β‐ODAP is oxidised by GlOx to form H₂O₂, which in turn is bioelectrocatalytically reduced by HRP through a mediated reaction using a polymeric mediator incorporating Os^{2 + /3+} functionalities rapidly shuttling electrons with the electrode_giving rise to the analytical signal. On the basis of this analysis system, the new isolates are capable of utilising β‐ODAP as sole carbon source to a maximum of 90–95% within 5 days with concomitant increase in cell protein. Copyright © 2005 Society of Chemical Industry     

Comparison of the performances of different fermentation strategies on cell growth and bacteriocin production by Lactobacillus curvatus CWBI‐B28

The dynamics of cell growth and bacteriocin production by Lactobacillus curvatus CWBI‐B28 in modified De Man/Rogosa/Sharp (mMRS) broth with various concentrations of glucose and complex nitrogen source (CNS; peptone, yeast extract and meat extract) was investigated in flask fermentations and in a laboratory fermentor using batch and fed‐batch cultivations. In fed‐batch fermentation the rate of feeding of the reactor with the substrates was either maintained constant (0.12 L h^?1) or varied exponentially as a function of time. The results showed that both cell growth and bacteriocin activity were influenced by changes in the concentrations of glucose and CNS. Optimal growth and bacteriocin activity were obtained in mMRS broth containing 40 g L^?1 glucose and 40 g L^?1 CNS (mMRS_40/40). A bacteriocin titre of 4266 AU mL^?1 and a cell count of 8.7 log colony‐forming units (cfu) mL^?1 were recorded when this medium was used for cultivation. In batch fermentation using the same medium, a higher cell count (9.5 log cfu mL^?1) and twice as much bacteriocin as in flask fermentation were produced. The highest bacteriocin titre (8533 AU mL^?1) was obtained with fed‐batch fermentation at an exponentially varying rate of feeding. Bacteriocin activity and cell dry mass did not always correlate. Copyright © 2007 Society of Chemical Industry     

冷榨花生粕蛋白胨酶法制备及其在微生物增殖培养中的应用

为探讨冷榨花生粕蛋白质酶法制备蛋白胨替代商用蛋白胨的可行性,将冷榨花生粕用胃蛋白酶与碱性蛋白酶进行双酶协同水解制备冷榨花生粕蛋白水解物（也称为冷榨花生粕蛋白胨）,并分析其分子质量分布、化学组分分析及氨基酸组成。研究发现,经双酶协同水解后的冷榨花生粕蛋白胨主要由小于5.0 kDa的肽段组成,富含丙氨酸、精氨酸、天冬氨酸、半胱氨酸等氨基酸,特别是谷氨酸含量较高。微生物增殖培养实验结果表明,大肠杆菌在冷榨花生粕蛋白胨培养基中培养8 h后比牛肉膏蛋白胨中生长情况好;枯草芽孢杆菌在冷榨花生粕蛋白胨培养基中比LB培养基中有较长的滞后期,但培养8 h后,生长情况比LB培养基更好;大肠杆菌和枯草芽孢杆菌在冷榨花生粕蛋白胨培养基和对照培养基中pH值变化差异不显著（P＞0.05）;干酪乳杆菌在冷榨花生粕蛋白胨培养基和MRS培养基中生长情况有显著差异（P＜0.05）;金黄色葡萄球菌在冷榨花生粕培养基中与对照培养基生长差异不显著（P＞0.05）;酿酒酵母在冷榨花生粕培养基中与对照生长有类似的生长曲线与pH值变化趋势;米曲霉在冷榨花生粕培养基中与对照培养生长无显著差异（P＞0.05）。结果表明,冷榨花生粕蛋白胨在替代商用蛋白胨用于微生物的增殖培养应用中具有很好的潜力。     

Biological denitrification in microbial fuel cells

Microbial fuel cells (MFCs) that remove carbon as well as nitrogen compounds out of wastewater are of special interest for practice. We developed a MFC in which microorganisms in the cathode performed a complete denitrification by using electrons supplied by microorganisms oxidizing acetate in the anode. The MFC with a cation exchange membrane was designed as a tubular reactor with an internal cathode and was able to remove up to 0.146 kg NO(3-)-N m(-3) net cathodic compartment (NCC) d(-1) (0.080 kg NO(3-)-N m(-3) total cathodic compartment d(-1) (TCC)) at a current of 58 A m(-3) NCC (32 A m(-3) TCC) and a cell voltage of 0.075 V. The highest power output in the denitrification system was 8 W m(-3) NCC (4 W m(-3) TCC) with a cell voltage of 0.214 V and a current of 35 A m(-3) NCC. The denitrification rate and the power production was limited bythe cathodic microorganisms, which only denitrified significantly at a cathodic electrode potential below 0 V versus standard hydrogen electrode (SHE). This is, to our knowledge, the first study in which a MFC has both a biological anode and cathode performing simultaneous removal of an organic substrate, power production, and complete denitrification without relying on H2-formation or external added power.     

Flocculation of the yeast Candida famata (Debaryomyces hansenii): An essential role for peptone

Aggregation of Candida famata (Debaryomyces hansenii) is consistent with being a form of lectin-mediated yeast flocculation. Flocculation of C. famata is unusual in that it requires the presence of peptone, either in the growth medium or added later to harvested cells in buffer. Flocculation after peptone addition was rapid, being largely complete within 10 min. Heat-killed cells also flocculated, arguing for direct participation of peptone in the flocculation binding mechanism. Flocculent C. famata cells progressively lost the ability to flocculate when washed with EDTA. Flocculation was fully restored by peptone addition; calcium addition was without effect. C. famata cells were able to agglutinate erythrocytes in the presence or absence of peptone. Pronase E-treated yeast lost both the ability to haemagglutinate and self-flocculate. Haemagglutination was not diminished by progressive EDTA washing, suggesting that surface lectins remained present and active on the yeast cell walls. Non-flocculating C. famata cells mutually flocculated with non-flocculent Schizosaccharomyces pombe cells, shown to have surface-exposed galactose residues. Mutual flocculation was lost following treatment of C. famata with Pronase E. It was concluded that the cell wall of C. famata contains lectins enabling haemagglutination and mutual flocculation but lacks carbohydrate receptors for these lectins. This yeast self-flocculates only via bridging multi-valent carbohydrates; these being present in peptone.     

阳电极面积对微生物燃料电池产电性能的影响

微生物燃料电池（MFC）最具应用前景之一是处理废水的同时能够产生电能。以糖蜜废水作为阳极基质,以金属离子的电镀废水做阴极溶液,研究了双室微生物燃料电池不同电极面积对产电性能和COD的影响。结果发现,当外电阻为300Q时,大反应器微生物燃料电池A．（阳极面积为78．15cm^2）及小反应器微生物燃料电池～（阳极面积为76．8cm^2）最大功率密度分别为0．28mW／cm^2和0．22mW／cm^2。在前200个小时内,A：电池在第60个小时时产生最大电压71．1mV和最大电流189．5μA,A,在第190个小时时产生最大电压81．1mV和最大电流228．1μA。同时,当Zn^2＋作阴极溶液时,小反应器微生物燃料电池阳极溶液的COD去除率在1．5％到7．02％之间,大反应器微生物燃料电池阳极溶液的COD去除率在0到14．96％之间。阴极中Zn^2＋去除率A1中为28．6％,A2为21．2％。     

Electricity-assisted biological hydrogen production from acetate by Geobacter sulfurreducens

Geobacter sulfurreducens is a well-known current-producing microorganism in microbial fuel cells, and is able to use acetate and hydrogen as electron donor. We studied the functionality of G. sulfurreducens as biocatalyst for hydrogen formation at the cathode of a microbial electrolysis cell (MEC). Geobacter sulfurreducens was grown in the bioelectrode compartment of a MFC with acetate as the substrate and reduction of complexed Fe(III) at the counter electrode. After depletion of the acetate the electrode potential of the bioelectrode was decreased stepwise to -1.0 V vs Ag/AgCl reference. Production of negative current was observed, which increased in time, indicating that the bioelectrode was now acting as biocathode. Headspace analyses carried out at electrode potentials ranging from -0.8 to -1.0 V showed that hydrogen was produced, with higher rates at more negative cathode potentials. Subsequently, the metabolic properties of G. sulfurreducens for acetate oxidation at the anode and hydrogen production at the cathode were combined in one-compartment membraneless MECs operated at applied voltages of 0.8 and 0.65 V. After two days, current densities were 0.44 A m(-2) at 0.8 V applied voltage and 0.22 A m(-2) at 0.65 V, using flat-surface carbon electrodes for both anode and cathode. The cathodic hydrogen recovery ranged from 23% at 0.5 V applied voltage to 43% at 0.9 V.     

Procedure for determining maximum sustainable power generated by microbial fuel cells

Power generated by microbial fuel cells is computed as a product of current passing through an external resistor and voltage drop across this resistor. If the applied resistance is very low, then high instantaneous power generated by the cell is measured, which is not sustainable; the cell cannot deliver that much power for long periods of time. Since using small electrical resistors leads to erroneous assessment of the capabilities of microbial fuel cells, a question arises: what resistor should be used in such measurements? To address this question, we have defined the sustainable power as the steady state of power delivery by a microbial fuel cell under a given set of conditions and the maximum sustainable power as the highest sustainable power that a microbial fuel cell can deliver under a given set of conditions. Selecting the external resistance that is associated with the maximum sustainable power in a microbial fuel cell (MFC) is difficult because the operator has limited influence on the main factors that control power generation: the rate of charge transfer at the current-limiting electrode and the potential established across the fuel cell. The internal electrical resistance of microbial fuel cells varies, and it depends on the operational conditions of the fuel cell. We have designed an empirical procedure to predict the maximum sustainable power that can be generated by a microbial fuel cell operated under a given set of conditions. Following the procedure, we change the external resistors incrementally, in steps of 500 omega every 10, 60, or 180 s and measure the anode potential, the cathode potential, and the cell current. Power generated in the microbial fuel cell that we were using was limited by the anodic current. The anodic potential was used to determine the condition where the maximum sustainable power is obtained. The procedure is simple, microbial fuel cells can be characterized within an hour, and the results of the measurements can serve many purposes, such as: (1) estimating power generation in various MFCs, (2) comparing power generation in MFCs using different electroactive reactants, (3) quantifying the effects of the operational regime on the power generation in MFCs, and finally, (4) the purpose for which the procedure was designed, optimizing the performance of existing MFCs.     

Growth optimization of thraustochytrid strain 12B for the commercial production of docosahexaenoic acid

High yield of cell, lipid, and docosahexaenoic acid (DHA) from thraustochytrid strain 12B were achieved without the use of a complex medium and at low NaCl concentration which is detrimental to avoid unnecessary corrosion of steel tank equipment during cultivation. Culture medium that contained only 0.1% NaCl and 1% MgSO₄ in an organic base solution containing 8% glucose, 1% yeast extract, and 1% peptone, referred here as NM medium, was found to be as good as or superior to the culture medium prepared from 50%(v/v) seawater with percentage lipid/dry cell weight (DCW) of 66.4%(w/w) and DHA yield up to 43.95 mg/g DCW for the thraustochytrid strain 12B. The NM medium was also applicable to the prominently high DHA-accumulating Schizochytrium limacinum SR21, and therefore this medium could probably be used for other thraustochytrid and other types of microbial strains as well.     

Screening of medium composition for the free radical‐scavenging properties by Antrodia cinnamomea

The effects of nutritional conditions on the scavenging capacity on superoxide anion radical and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) free radical of ethanolic extracts from filtrates of Antrodia cinnamomea were investigated in shake‐flask cultures. Culture medium significantly affected inhibition of superoxide anion generation and DPPH free radical scavenging activity. In contrast, both scavenging capabilities on superoxide anion radical and DPPH free radical were found to be greatly affected by varying the carbon source, the nitrogen source, the growth factors or the trace elements. The maximal inhibition of superoxide anion generation could be obtained when the culture medium compositions were: xylose, ammonium oxalate, nicotinic acid, NaH₂PO₄ and CuSO₄; while a maximal DPPH free radical scavenging activity could be achieved when the medium composition setting as: fructose, peptone, nicotinic acid, KH₂PO₄ and CaCl₂ (the scavenging effects on superoxide anion radical and DPPH free radical were increased to 96.3 ± 0.3% and 29.0 ± 1.0%, respectively). We proved that controlling the culturing conditions and modifying the composition of the medium can dramatically enhance the scavenging ability on superoxide anion radical and DPPH free radical of A. cinnamomea.     

Scaling up microbial fuel cells

The goal of this study was to quantify the relation between the surface area of the current-limiting electrode of a microbial fuel cell (MFC) and the power density generated by the MFC. Shewanella oneidensis (MR-1) was grown anaerobically in the anodic compartment of an MFC utilizing lactate as the electron donor. Graphite plate electrodes of various sizes were used as anodes. Commercially available air electrodes, composed of manganese-based catalyzed carbon bonded to a current-collecting screen made of platinum mesh, were used as cathodes, and dissolved oxygen was used as the cathodic reactant. The surface area of the cathode was always significantly larger than that of the anode, to ensure that the anode was the current-limiting electrode. The power density generated by the MFC decreased as the surface area of the anode increased, which fits well with the trend we detected comparing various published results. Thus, our findings bring into question the assertion that the overall power density generated by an MFC with large electrodes can be estimated by extrapolating from an electrode with a small surface area. Our results indicate that the maximum power density generated by an MFC is not directly proportional to the surface area of the anode, but is instead proportional to the logarithm of the surface area of the anode.     

Microbial succession and metabolite changes during the fermentation of Chinese light aroma‐style liquor

A combination of culture‐dependent and culture‐independent methods and SPME–GC–MS were used to monitor changes of bacterial and yeast communities, and flavour compounds during the fermentation process of Chinese light aroma‐style liquor. Bacillus and Lactobacillus were the main bacterial genera. Pichia anomala, Saccharomyces cerevisiae and Issatchenkia orientalis were the dominant yeast species. There was a close relationship between fermentation time and the shift of microbial community. Compared with the microbiota in the fermentation of other style liquors, higher bacterial diversity and different non‐Saccharomyces composition led to a variety of metabolites. Metabolite analysis showed that esters, acids, alcohols, aromatic compounds and phenols were the main flavour components and most of them were synthesised in the latter phase of fermentation. Principal component analysis further demonstrated that Bacillus and yeast were the most influential microorganisms in the first 10 days of fermentation, and lactic acid bacteria predominated in the later phase. Lactic acid bacteria regulated the composition of other bacteria and yeast, and synthesised flavour compounds to affect the organoleptic properties of liquor. S. cerevisiae and P. anomala were two important yeast species responsible for the characteristic aroma of liquor. These results present a comprehensive understanding of microbial interaction and potential starter cultures to produce desirable liquor quality. © 2018 The Institute of Brewing & Distilling     

Overexpression of vacuolar H+‐ATPase‐related genes in bottom‐fermenting yeast enhances ethanol tolerance and fermentation rates during high‐gravity fermentation

Vacuolar H⁺‐ATPase (V‐ATPase) is thought to play a role in stress tolerance. In this study it was found that bottom‐fermenting yeast strains, in which the V‐ATPase‐related genes DBF2, VMA41/CYS4/NHS5 and RAV2 were overexpressed, exhibited stronger ethanol tolerance than the parent strain and showed increased fermentation rates in a high‐sugar medium simulating high‐gravity fermentation. Among the strains examined, the DBF2‐overexpressing bottom‐fermenting yeast strain exhibited the highest ethanol tolerance and fermentation rate in YPM20 medium. Using this strain, high‐gravity fermentation was performed by adding sugar to the wort, which led to increased fermentation rates and yeast viability compared with the parent strain. These findings indicate that V‐ATPase is a stress target in high‐gravity fermentation and suggests that enhancing the V‐ATPase activity increases the ethanol tolerance of bottom‐fermenting yeast, thereby improving the fermentation rate and cell viability under high‐gravity conditions. Copyright © 2012 The Institute of Brewing & Distilling     

Inverse Flocculation Patterns in Saccharomyces cerevisiae UOFS Y‐2330

An interesting yeast strain was uncovered which showed an inverse flocculation pattern when cultivated in chemically defined and complex media. When inoculated in a defined medium with glucose as a sole carbon source, this strain immediately flocculated strongly and lost this ability before stationary phase was reached. In a complex malt medium containing glucose, this yeast strongly flocculated throughout the exponential and stationary growth phases. This inverse pattern may be ascribed to a switch in sensitivity of the yeast to flocculate in the presence of glucose as well as pH level, which may, in turn, influence the availability of calcium ions. In both media, matured cells produced protuberances or “wrinkles” upon flocculation as observed by electron and immunofluorescence microscopy. These protuberances may be involved in cell adhesion during the flocculation process.