Microbial Catalysts Research Articles

The decisive role of acidophilic bacteria in concrete sewer networks: A new model for fast progressing microbial concrete corrosion

This study introduces a novel approach intertwining analytics of spatial microbial distribution with chemical, mineralogical and (micro)structural related aspects in corroded concrete sewer environments. Samples containing up to 4cm thick corrosion layers were collected from concrete manholes and analysed using hydro-geochemical, microbiological, biochemical and mineralogical methods. Opposed to the current opinion DNA and RNA indicating microbial activity were found throughout the entire deterioration layer down to the corrosion front. Elemental distributions of corresponding areas revealed a dynamic pH- and diffusion-controlled system in which a distinct succession of elemental accumulations was unequivocally correlated with responding pH levels, associated dissolution and precipitation of solids, as well as with the spatially resolved presence of microbes. Microbial activity further coincided with massive iron deposition zones, within the inner anoxic to anaerobic corrosion layers. As a possible microbial catalyst for iron oxidation and in-situ acid production in this zone, we propose Acidithiobacillus ferrooxidans which were isolated from the deteriorated concrete. Based on the data we propose a new model in which biogenic induced in-situ acid production is a decisive factor, steering high concrete corrosion rates of >1cmyr−1.

Continuous biogenic hydrogen production from dilute acid pretreated algal hydrolysate using hybrid immobilized mixed consortia

This study investigated the bioconversion of dilute acid (2% H2SO4) pretreated red algae (Gelidium amansii) hydrolysate into H2 by anaerobic fermentation in a continuous stirred tank reactor under mesophilic conditions using hybrid immobilized cells as microbial catalyst. Two different hydraulic retention times (HRT) of 24 h and 16 h with a feed concentration of 15 g/L hexose equivalent have been investigated over 85 days of operation to evaluate H2 production performance and stability of the continuous system. The highest hydrogen production rate (HPR) and hydrogen yield (HY) of 2.7 L/L/d and 1.3 mol/mol substrate hexoseadded was achieved at 24 h HRT, while further operation at 16 h HRT led to a significant drop in the hydrogen production with a HPR and HY values of 1.8 L/L/d and 0.7 mol/mol substrate hexoseadded, respectively. The bacterial community analysis characterized by 454 pyrosequencing revealed that the changes in HRT significantly influence the composition of the dominant microflora. At longer HRT (24 h), the phyla Firmicutes was abundant over 98%, whereas at shorter HRT (16 h), Proteobacteria being the dominant populations with 84%. These outcomes suggested that controlling appropriate HRT is prerequisite for efficient hydrogen production.

Microbial Electrosynthesis I: Pure and Defined Mixed Culture Engineering.

In the past 6years, microbial bioelectrochemistry has strongly increased in attraction and audience when expanding from mainly environmental technology applications to biotechnology. In particular, the promise to combine electrosynthesis with microbial catalysis opens attractive approaches for new sustainable redox-cofactor recycling, redox-balancing, or even biosynthesis processes. Much of this promise is still not fulfilled, but it has opened and fueled entirely new research areas in this discipline. Activities in designing, tailoring, and applying specific microbial catalysts as pure or defined co-cultures for defined target bioproductions are greatly accelerating. This chapter gives an overview of the current progress as well as the emerging trends in molecular and ecological engineering of defined microbial biocatalysts to prepare them for evolving microbial electrosynthesis processes. In addition, the multitude of microbial electrosynthetic processes with complex undefined mixed cultures is covered by ter Heijne et al. (Adv Biochem Eng Biotechnol. https://doi.org/10.1007/10_2017_15 , 2017). Graphical Abstract.

The effect of anode potential on bioelectrochemical and electrochemical tetrathionate degradation

The effect of poised anode potential on electricity production and tetrathionate degradation was studied in two-chamber flow-through electrochemical (ES) and bioelectrochemical systems (BES). The minimum anode potential (vs. Ag/AgCl) for positive current generation was 0.3V in BES and 0.5V in the abiotic ES. The anode potential required to obtain average current density above 70mAm−2 was 0.4V in BES and above 0.7V in ES. ES provided higher coulombic efficiency, but the average tetrathionate degradation rate remained significantly higher in BES (above 110mgL−1d−1) than in the abiotic ES (below 35mgL−1d−1). This study shows that at anode potentials below 0.7V, the electrochemical tetrathionate degradation is only efficient with microbial catalyst and that significantly higher tetrathionate degradation rates can be obtained with bioelectrochemical systems than with electrochemical systems at the tested anode potentials.

Yeast and carbon nanotube based biocatalyst developed by synergetic effects of covalent bonding and hydrophobic interaction for performance enhancement of membraneless microbial fuel cell

Membraneless microbial fuel cell (MFC) employing new microbial catalyst formed as yeast cultivated from Saccharomyces cerevisiae and carbon nanotube (yeast/CNT) is suggested. To analyze its catalytic activity and performance and stability of MFC, several characterizations are performed. According to the characterizations, the catalyst shows excellent catalytic activities by facile transfer of electrons via reactions of NAD, FAD, cytochrome c and cytochrome a3, while it induces high maximum power density (MPD) (344mW·m−2). It implies that adoption of yeast induces increases in catalytic activity and MFC performance. Furthermore, MPD is maintained to 86% of initial value even after eight days, showing excellent MFC stability.

Enhanced production of insect raw-starch-digesting alpha-amylase accompanied by high erythritol synthesis in recombinant Yarrowia lipolytica fed-batch cultures at high-cell-densities

Abstract Recently we reported on raw-starch-digesting ability of alpha-amylase from an insect Sitophilus oryzae (SoAMY) expressed in recombinant Yarrowia lipolytica cells, and demonstrated its usefulness in simultaneous saccharification and fermentation processes with industrial yeasts. In this study we applied fed-batch cultures of Y. lipolytica 4.29 strain reaching high-cell-densities (up to 70 [gDCW/L]), to enhance SoAMY production. SoAMY activity in the medium reached the peak value of 22,979.23 ± 184 [AU/L], at volumetric productivity of 121.58 ± 1.75 [AU/L/h], and yield of 71.83 ± 3.08 [AU/gglycerol], constituting roughly 160-fold improvement, compared to the best previous result. The cultivations were accompanied by high production of erythritol (83.58 [g/L]), at the marginal production of mannitol (5.46 [g/L]). Elementary analyses of media constituents, the enzyme and the yeast biomass gave better insight into carbon and nitrogen fluxes distribution. Due to application of genetic engineering and bioprocess engineering strategies, the insect-derived enzyme can be produced at the quantities competitive to microbial catalysts.

Enhanced microbial electrosynthesis with three-dimensional graphene functionalized cathodes fabricated via solvothermal synthesis

The biological reduction of CO2 into multicarbon chemicals can be driven by electrons derived from the cathode of a bioelectrochemical reactor via microbial electrosynthesis (MES). To increase MES productivity, conditions for optimal electron transfer between the cathode and the microbial catalyst must be implemented. Here, we report the development of a 3D-graphene functionalized carbon felt composite cathode enabling faster electron transfer to the microbial catalyst Sporomusa ovata in a MES reactor. Modification with 3D-graphene network increased the electrosynthesis rate of acetate from CO2 by 6.8 fold. It also significantly improved biofilm density and current consumption. A 2-fold increase in specific surface area of the 3D-graphene/carbon felt composite cathode explained in part the formation of more substantial biofilms compared to untreated control. Furthermore, in cyclic voltammetry analysis, 3D-graphene/carbon felt composite cathode exhibited higher current response. The results indicate that the development of a 3D-network cathode is an effective approach to improve microbe-electrode interactions leading to productive MES systems.

Effect of tungstate on acetate and ethanol production by the electrosynthetic bacterium Sporomusa ovata.

BackgroundMicrobial electrosynthesis (MES) and gas fermentation are bioenergy technologies in which a microbial catalyst reduces CO2 into organic carbon molecules with electrons from the cathode of a bioelectrochemical system or from gases such as H2. The acetogen Sporomusa ovata has the capacity of reducing CO2 into commodity chemicals by both gas fermentation and MES. Acetate is often the only product generated by S. ovata during autotrophic growth.ResultsIn this study, trace elements in S. ovata growth medium were optimized to improve MES and gas fermentation productivity. Augmenting tungstate concentration resulted in a 2.9-fold increase in ethanol production by S. ovata during H2:CO2-dependent growth. It also promoted electrosynthesis of ethanol in a S. ovata-driven MES reactor and increased acetate production 4.4-fold compared to unmodified medium. Furthermore, fatty acids propionate and butyrate were successfully converted to their corresponding alcohols 1-propanol and 1-butanol by S. ovata during gas fermentation. Increasing tungstate concentration enhanced conversion efficiency for both propionate and butyrate. Gene expression analysis suggested that tungsten-containing aldehyde ferredoxin oxidoreductases (AORs) and a tungsten-containing formate dehydrogenase (FDH) were involved in the improved biosynthesis of acetate, ethanol, 1-propanol, and 1-butanol. AORs and FDH contribute to the fatty acids re-assimilation pathway and the Wood–Ljungdahl pathway, respectively.ConclusionsThis study presented here shows that optimization of microbial catalyst growth medium can improve productivity and lead to the biosynthesis of different products by gas fermentation and MES. It also provides insights on the metabolism of biofuels production in acetogens and demonstrates that S. ovata has an important untapped metabolic potential for the production of other chemicals than acetate via CO2-converting bioprocesses including MES.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0576-0) contains supplementary material, which is available to authorized users.

Hierarchically Three-Dimensional Nanofiber Based Textile with High Conductivity and Biocompatibility As a Microbial Fuel Cell Anode.

Microbial fuel cells (MFCs) encompass complex bioelectrocatalytic reactions that converting chemical energy of organic compounds to electrical energy. Improving the anode configuration is thought to be a critical step for enhancing MFCs performance. In present study, a hierarchically structured textile polypyrrole/poly(vinyl alcohol-co-polyethylene) nanofibers/poly(ethylene terephthalate) (referred to PPy/NFs/PET) is shown to be excellent anode for MFCs. This hierarchical PPy/NFs/PET anode affords an open porous and three-dimensional interconnecting conductive scaffold with larger surface roughness, facilitating microbial colonization and electron transfer from exoelectrogens to the anode. The mediator-less MFC equipped with PPy/NFs/PET anode achieves a remarkable maximum power density of 2420 mW m(-2) with Escherichia coli as the microbial catalyst at the current density of 5500 mA m(-2), which is approximately 17 times higher compared to a reference anode PPy/PET (144 mW m(-2)). Considering the low cost, low weight, facile fabrication, and good winding, this PPy/NFs/PET textile anode promises a great potential for high-performance and cost-effective MFCs in a large scale.

Editorial: Current Challenges and Future Perspectives on Emerging Bioelectrochemical Technologies.

Editorial: Current Challenges and Future Perspectives on Emerging Bioelectrochemical Technologies.

Enhanced power generation using a novel polymer-coated nanoparticles dispersed-carbon micro-nanofibers-based air-cathode in a membrane-less single chamber microbial fuel cell

A novel polymer-metal nanoparticle-carbon micro-nanofiber-based nanocomposite was developed as an efficient air-cathode of a membrane-less single chamber microbial fuel cell (SMFC) for bio-energy production using Escherichia coli as a microbial catalyst. The polymeric material was synthesized using suspension polymerization, with polyvinyl alcohol as a monomer and poly methyl vinyl ether-alt-maleic anhydride (PMVEMA) as a cross-linking agent. The synthesized polymer composite was coated over the alumina (AA) and nickel (Ni) nanoparticles (NPs)-dispersed multi-scale web of activated carbon fiber (ACF) and carbon nanofiber (CNF). The PMVEMA polymer facilitated proton exchange from anolyte to cathode in SMFCs. AA:Ni-ACF/CNF served as an electron acceptor as well as the catalyst for atmospheric oxygen reduction at the cathode. The polarization curves determined using linear sweep voltammetry demonstrated the prepared polymer-metal-carbon nanocomposite to be an efficient air-cathode of the SMFC with a maximum electrical potential of 980 ± 10 mV and a maximum power density of 1270 ± 30 mW/m2.

Identification of Genes Conferring Tolerance to Lignocellulose-Derived Inhibitors by Functional Selections in Soil Metagenomes.

The production of fuels or chemicals from lignocellulose currently requires thermochemical pretreatment to release fermentable sugars. These harsh conditions also generate numerous small-molecule inhibitors of microbial growth and fermentation, limiting production. We applied small-insert functional metagenomic selections to discover genes that confer microbial tolerance to these inhibitors, identifying both individual genes and general biological processes associated with tolerance to multiple inhibitory compounds. Having screened over 248 Gb of DNA cloned from 16 diverse soil metagenomes, we describe gain-of-function tolerance against acid, alcohol, and aldehyde inhibitors derived from hemicellulose and lignin, demonstrating that uncultured soil microbial communities hold tremendous genetic potential to address the toxicity of pretreated lignocellulose. We recovered genes previously known to confer tolerance to lignocellulosic inhibitors as well as novel genes that confer tolerance via unknown functions. For instance, we implicated galactose metabolism in overcoming the toxicity of lignin monomers and identified a decarboxylase that confers tolerance to ferulic acid; this enzyme has been shown to catalyze the production of 4-vinyl guaiacol, a valuable precursor to vanillin production. These metagenomic tolerance genes can enable the flexible design of hardy microbial catalysts, customized to withstand inhibitors abundant in specific bioprocessing applications.

Adaptation of the autotrophic acetogen Sporomusa ovata to methanol accelerates the conversion of CO2 to organic products.

Acetogens are efficient microbial catalysts for bioprocesses converting C1 compounds into organic products. Here, an adaptive laboratory evolution approach was implemented to adapt Sporomusa ovata for faster autotrophic metabolism and CO2 conversion to organic chemicals. S. ovata was first adapted to grow quicker autotrophically with methanol, a toxic C1 compound, as the sole substrate. Better growth on different concentrations of methanol and with H2-CO2 indicated the adapted strain had a more efficient autotrophic metabolism and a higher tolerance to solvent. The growth rate on methanol was increased 5-fold. Furthermore, acetate production rate from CO2 with an electrode serving as the electron donor was increased 6.5-fold confirming that the acceleration of the autotrophic metabolism of the adapted strain is independent of the electron donor provided. Whole-genome sequencing, transcriptomic, and biochemical studies revealed that the molecular mechanisms responsible for the novel characteristics of the adapted strain were associated with the methanol oxidation pathway and the Wood-Ljungdahl pathway of acetogens along with biosynthetic pathways, cell wall components, and protein chaperones. The results demonstrate that an efficient strategy to increase rates of CO2 conversion in bioprocesses like microbial electrosynthesis is to evolve the microbial catalyst by adaptive laboratory evolution to optimize its autotrophic metabolism.

Possibilities for extremophilic microorganisms in microbial electrochemical systems.

Microbial electrochemical systems exploit the metabolism of microorganisms to generate electrical energy or a useful product. In the past couple of decades, the application of microbial electrochemical systems has increased from the use of wastewaters to produce electricity to a versatile technology that can use numerous sources for the extraction of electrons on the one hand, while on the other hand these electrons can be used to serve an ever increasing number of functions. Extremophilic microorganisms grow in environments that are hostile to most forms of life and their utilization in microbial electrochemical systems has opened new possibilities to oxidize substrates in the anode and produce novel products in the cathode. For example, extremophiles can be used to oxidize sulfur compounds in acidic pH to remediate wastewaters, generate electrical energy from marine sediment microbial fuel cells at low temperatures, desalinate wastewaters and act as biosensors of low amounts of organic carbon. In this review, we will discuss the recent advances that have been made in using microbial catalysts under extreme conditions and show possible new routes that extremophilic microorganisms open for microbial electrochemical systems.

Metabolic and microbial community dynamics during the anaerobic digestion of maize silage in a two-phase process.

Two-phasic anaerobic digestion processes (hydrolysis/acidogenesis separated from acetogenesis/methanogenesis) can be used for biogas production on demand or a combined chemicals/bioenergy production. For an effective process control, detailed knowledge about the microbial catalysts and their correlation to process conditions is crucial. In this study, maize silage was digested in a two-phase process and interrelationships between process parameters and microbial communities were revealed. In the first-phase reactor, alternating metabolic periods were observed which emerged independently from the feeding frequency. During the L-period, up to 11.8gL(-1) lactic acid was produced which significantly correlated to lactic acid bacteria of the genus Lactobacillus as the most abundant community members. During the alternating G-period, the production of volatile fatty acids (up to 5.3, 4.0 and 3.1gL(-1) for propionic, n-butyric and n-caproic acid, respectively) dominated accompanied by a high gas production containing up to 28% hydrogen. The relative abundance of various Clostridiales increased during this metabolic period. In the second-phase reactor, the metabolic fluctuations of the first phase were smoothed out resulting in a stable biogas production as well as stable bacterial and methanogenic communities. However, the biogas composition followed the metabolic dynamics of the first phase: the hydrogen content increased during the L-period whereas highest CH4/CO2 ratios (up to 2.8) were reached during the G-period. Aceticlastic Methanosaeta as well as hydrogenotrophic Methanoculleus and Methanobacteriaceae were identified as dominant methanogens. Consequently, a directed control of the first-phase stabilizing desired metabolic states can lead to an enhanced productivity regarding chemicals and bioenergy.

Metabolic engineering of Escherichia coli into a versatile glycosylation platform: production of bio-active quercetin glycosides.

BackgroundFlavonoids are bio-active specialized plant metabolites which mainly occur as different glycosides. Due to the increasing market demand, various biotechnological approaches have been developed which use Escherichia coli as a microbial catalyst for the stereospecific glycosylation of flavonoids. Despite these efforts, most processes still display low production rates and titers, which render them unsuitable for large-scale applications.ResultsIn this contribution, we expanded a previously developed in vivo glucosylation platform in E. coli W, into an efficient system for selective galactosylation and rhamnosylation. The rational of the novel metabolic engineering strategy constitutes of the introduction of an alternative sucrose metabolism in the form of a sucrose phosphorylase, which cleaves sucrose into fructose and glucose 1-phosphate as precursor for UDP-glucose. To preserve these intermediates for glycosylation purposes, metabolization reactions were knocked-out. Due to the pivotal role of UDP-glucose, overexpression of the interconverting enzymes galE and MUM4 ensured the formation of both UDP-galactose and UDP-rhamnose, respectively. By additionally supplying exogenously fed quercetin and overexpressing a flavonol galactosyltransferase (F3GT) or a rhamnosyltransferase (RhaGT), 0.94 g/L hyperoside (quercetin 3-O-galactoside) and 1.12 g/L quercitrin (quercetin 3-O-rhamnoside) could be produced, respectively. In addition, both strains showed activity towards other promising dietary flavonols like kaempferol, fisetin, morin and myricetin.ConclusionsTwo E. coli W mutants were engineered that could effectively produce the bio-active flavonol glycosides hyperoside and quercitrin starting from the cheap substrates sucrose and quercetin. This novel fermentation-based glycosylation strategy will allow the economically viable production of various glycosides.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-015-0326-1) contains supplementary material, which is available to authorized users.

폐광산지역 토양 식생복원 과정 내 토양특성 및 미생물 군집 변화 분석

Mine soil contamination by high levels of metal ions that prevents the successful vegetation poses a serious problem. In the study presented here, we used the microbial biocatalyst of urease producing bacterium Sporosarcina pasteurii or plant extract based BioNeutro-GEM (BNG) agent. The ability of the biocatalysts to bioremediate contaminated soil from abandoned mine was examined by solid-state composting vegetation under field conditions. Treatment of mine soil with the 2 biocatalysts for 5 months resulted in pH increase and electric conductivity reduction compared to untreated control. Further analyses revealed that the microbial catalysts also promoted the root and shoot growth to the untreated control during the vegetation treatments. After the Sporosarcina pasteurii or plant extract based BNG treatment, the microbial community change was monitored by culture-independent pyrosequencing. These results demonstrate that the microbial biocatalysts could potentially be used in the soil bioremediation from mine-impacted area.

Metabolic modeling of synthesis gas fermentation in bubble column reactors.

BackgroundA promising route to renewable liquid fuels and chemicals is the fermentation of synthesis gas (syngas) streams to synthesize desired products such as ethanol and 2,3-butanediol. While commercial development of syngas fermentation technology is underway, an unmet need is the development of integrated metabolic and transport models for industrially relevant syngas bubble column reactors.ResultsWe developed and evaluated a spatiotemporal metabolic model for bubble column reactors with the syngas fermenting bacterium Clostridium ljungdahlii as the microbial catalyst. Our modeling approach involved combining a genome-scale reconstruction of C. ljungdahlii metabolism with multiphase transport equations that govern convective and dispersive processes within the spatially varying column. The reactor model was spatially discretized to yield a large set of ordinary differential equations (ODEs) in time with embedded linear programs (LPs) and solved using the MATLAB based code DFBAlab. Simulations were performed to analyze the effects of important process and cellular parameters on key measures of reactor performance including ethanol titer, ethanol-to-acetate ratio, and CO and H2 conversions.ConclusionsOur computational study demonstrated that mathematical modeling provides a complementary tool to experimentation for understanding, predicting, and optimizing syngas fermentation reactors. These model predictions could guide future cellular and process engineering efforts aimed at alleviating bottlenecks to biochemical production in syngas bubble column reactors.

Microbial electron transport and energy conservation - the foundation for optimizing bioelectrochemical systems.

Microbial electrochemical techniques describe a variety of emerging technologies that use electrode–bacteria interactions for biotechnology applications including the production of electricity, waste and wastewater treatment, bioremediation and the production of valuable products. Central in each application is the ability of the microbial catalyst to interact with external electron acceptors and/or donors and its metabolic properties that enable the combination of electron transport and carbon metabolism. And here also lies the key challenge. A wide range of microbes has been discovered to be able to exchange electrons with solid surfaces or mediators but only a few have been studied in depth. Especially electron transfer mechanisms from cathodes towards the microbial organism are poorly understood but are essential for many applications such as microbial electrosynthesis. We analyze the different electron transport chains that nature offers for organisms such as metal respiring bacteria and acetogens, but also standard biotechnological organisms currently used in bio-production. Special focus lies on the essential connection of redox and energy metabolism, which is often ignored when studying bioelectrochemical systems. The possibility of extracellular electron exchange at different points in each organism is discussed regarding required redox potentials and effect on cellular redox and energy levels. Key compounds such as electron carriers (e.g., cytochromes, ferredoxin, quinones, flavins) are identified and analyzed regarding their possible role in electrode–microbe interactions. This work summarizes our current knowledge on electron transport processes and uses a theoretical approach to predict the impact of different modes of transfer on the energy metabolism. As such it adds an important piece of fundamental understanding of microbial electron transport possibilities to the research community and will help to optimize and advance bioelectrochemical techniques.

Simplifying microbial electrosynthesis reactor design.

Microbial electrosynthesis, an artificial form of photosynthesis, can efficiently convert carbon dioxide into organic commodities; however, this process has only previously been demonstrated in reactors that have features likely to be a barrier to scale-up. Therefore, the possibility of simplifying reactor design by both eliminating potentiostatic control of the cathode and removing the membrane separating the anode and cathode was investigated with biofilms of Sporomusa ovata. S. ovata reduces carbon dioxide to acetate and acts as the microbial catalyst for plain graphite stick cathodes as the electron donor. In traditional ‘H-cell’ reactors, where the anode and cathode chambers were separated with a proton-selective membrane, the rates and columbic efficiencies of microbial electrosynthesis remained high when electron delivery at the cathode was powered with a direct current power source rather than with a potentiostat-poised cathode utilized in previous studies. A membrane-less reactor with a direct-current power source with the cathode and anode positioned to avoid oxygen exposure at the cathode, retained high rates of acetate production as well as high columbic and energetic efficiencies. The finding that microbial electrosynthesis is feasible without a membrane separating the anode from the cathode, coupled with a direct current power source supplying the energy for electron delivery, is expected to greatly simplify future reactor design and lower construction costs.