Syngas Fermentation Research Articles

Direct Medium-Chain Carboxylic Acid Oil Separation from a Bioreactor by an Electrodialysis/Phase Separation Cell.

Medium-chain carboxylic acids (MCCAs) are valuable platform chemicals and can be produced from waste biomass sources or syngas fermentation effluent through microbial chain elongation. We have previously demonstrated successful approaches to separate >90% purity oil with different MCCAs (MCCA oil) by integrating the anaerobic bioprocess with membrane-based liquid-liquid extraction (pertraction) and membrane electrolysis. However, two-compartment membrane electrolysis unit without pertraction was not able to separate MCCA oil. Therefore, we developed a five-compartment electrodialysis/phase separation cell (ED/PS). First, we tested an ED/PS cell in series with pertraction and achieved a maximum MCCA-oil flux of 1.7 × 103 g d-1 per projected area (m2) (19 mL oil d-1) and MCCA-oil transfer efficiency [100% × moles MCCA-oil moles electrons-1] of 74% at 15 A m-2. This extraction system at 15 A m-2 demonstrated a ∼10 times lower electric-power consumption (1.1 kWh kg-1 MCCA oil) than membrane electrolysis in series with pertraction (9.9 kWh kg-1 MCCA oil). Second, we evaluated our ED/PS as a stand-alone unit when integrated with the anaerobic bioprocess and demonstrated that we can selectively extract and separate MCCA oil directly from chain-elongating bioreactor broth with just an abiotic electrochemical cell. However, the electric-power consumption increased considerably due to the lower MCCA concentrations in the bioreactor broth compared to the pertraction broth.

Methanol supply speeds up synthesis gas fermentation by methylotrophic-acetogenic bacterium, Eubacterium limosum KIST612

This study analyzed the effect of methanol on the metabolism of syngas components (i.e., H2 and CO) by the syngas fermenting acetogenic strain E. limosum KIST612. The culture characteristics and relevant proteomic expressions (as fold changes) were carefully analyzed under CO/CO2 and H2/CO2 conditions with and without methanol addition, as well as, under methanol/CO2 conditions. The culture characteristics (specific growth rate and H2 consumption rate) under H2/CO2 conditions were greatly enhanced in the presence of methanol, by 4.0 and 2.7 times, respectively. However, the promoting effect of methanol was not significant under CO/CO2 conditions. Proteomic fold changes in most enzyme expression levels in the Wood-Ljungdahl pathway and chemiosmotic energy conservation also exhibited high correspondence between methanol and H2/CO2 but not between methanol and CO/CO2. These findings suggest the advantages of methanol addition to H2/CO2 for biomass enhancement and faster consumption of gaseous substrates during syngas fermentation.

Syngas Derived from Lignocellulosic Biomass Gasification as an Alternative Resource for Innovative Bioprocesses

A hybrid system based on lignocellulosic biomass gasification and syngas fermentation represents a second-generation biorefinery approach that is currently in the development phase. Lignocellulosic biomass can be gasified to produce syngas, which is a gas mixture consisting mainly of H2, CO, and CO2. The major challenge of biomass gasification is the syngas’s final quality. Consequently, the development of effective syngas clean-up technologies has gained increased interest in recent years. Furthermore, the bioconversion of syngas components has been intensively studied using acetogenic bacteria and their Wood–Ljungdahl pathway to produce, among others, acetate, ethanol, butyrate, butanol, caproate, hexanol, 2,3-butanediol, and lactate. Nowadays, syngas fermentation appears to be a promising alternative for producing commodity chemicals in comparison to fossil-based processes. Research studies on syngas fermentation have been focused on process design and optimization, investigating the medium composition, operating parameters, and bioreactor design. Moreover, metabolic engineering efforts have been made to develop genetically modified strains with improved production. In 2018, for the first time, a syngas fermentation pilot plant from biomass gasification was built by LanzaTech Inc. in cooperation with Aemetis, Inc. Future research will focus on coupling syngas fermentation with additional bioprocesses and/or on identifying new non-acetogenic microorganisms to produce high-value chemicals beyond acetate and ethanol.

Influence of Liquid-to-Gas Ratio on the Syngas Fermentation Efficiency: An Experimental Approach.

Syngas fermentation by methanogens is a novel process to purify biogas. Methanogens are able to ferment non-desirable CO2, H2, and CO to methane. However, to use methanogens on an industrial scale, more research has to be done. There are studies that discuss the growth of methanogens on syngas in combination with acetate. In this research, growth of methanogens on syngas as sole carbon source is discussed. Effluent of an anaerobic fed-batch was selectively cultivated with syngas in 400 mL Eppendorf© bioreactors. After a period of 7 days, fifteen 120 mL flasks were filled with three different liquid-to-gas ratios (1:1, 1:3, 1:5). Results showed that different liquid-to-gas ratios change the metabolic preference of the anaerobic microbial community. Moreover, complete conversion in a four-to-eight-day period, via the carboxidotrophic pathway, was observed in all three liquid-to-gas ratios.

Effect of temperature and surfactant on biomass growth and higher-alcohol production during syngas fermentation by Clostridium carboxidivorans P7

Hexanol–butanol–ethanol fermentation from syngas by Clostridium carboxidivorans P7 is a promising route for biofuel production. However, bacterial agglomeration in the culture of 37 °C severely hampers the accumulation of biomass and products. To investigate the effect of culture temperature on biomass growth and higher-alcohol production, C. carboxidivorans P7 was cultivated at both constant and two-step temperatures in the range from 25 to 37 °C. Meanwhile, Tween-80 and saponin were screened out from eight surfactants to alleviate agglomeration at 37 °C. The results showed that enhanced higher-alcohol production was contributed mainly by the application of two-step temperature culture rather than the addition of anti-agglomeration surfactants. Furthermore, comparative transcriptome revealed that although 37 °C promoted high expression of genes involved in the Wood–Ljungdahl pathway, genes encoding enzymes catalyzing acyl-condensation reactions associated with higher-alcohol production were highly expressed at 25 °C. This study gained greater insight into temperature-effect mechanism on syngas fermentation by C. carboxidivorans P7.

Biochar and activated carbon enhance ethanol conversion and selectivity to caproic acid by Clostridium kluyveri

Syngas from biomass or steel mills can be fermented into a dilute stream of ethanol and acetic acid, which requires energy intensive distillation for product recovery. This can be circumvented by selective secondary fermentation of the syngas fermentation effluent to caproic acid as easier recoverable platform chemical with Clostridium kluyveri. Here, we explore the impact of biochar and activated carbon on this process. Changes during the fermentation with biochar or activated carbon were monitored, different doses were tested and the recyclability of biochar and activated carbon was assessed. Biochar decreased the lag phase and increased the caproic acid production rate (up to 0.50 g·L-1·h-1). Upon recycling for subsequent fermentation, biochar retained this property largely. Activated carbon addition, especially at high dose, could potentially increase the conversion and selectivity towards caproic acid to 14.15 g·L-1 (control: 11.01 g·L-1) and 92% (control: 84%), respectively.

Online measurement of dissolved carbon monoxide concentrations reveals critical operating conditions in gas fermentation experiments

Syngas fermentation is one possible contributor to the reduction of greenhouse gas emissions. The conversion of industrial waste gas streams containing CO or H2 , which are usually combusted, directly reduces the emission of CO2 into the atmosphere. Additionally, other carbon-containing waste streams can be gasified, making them accessible for microbial conversion into platform chemicals. However, there is still a lack of detailed process understanding, as online monitoring of dissolved gas concentrations is currently not possible. Several studies have demonstrated growth inhibition of Clostridium ljungdahlii at high CO concentrations in the headspace. However, growth is not inhibited by the CO concentration in the headspace, but by the dissolved carbon monoxide tension (DCOT). The DCOT depends on the CO concentration in the headspace, CO transfer rate, and biomass concentration. Hence, the measurement of the DCOT is a superior method to investigate the toxic effects of CO on microbial fermentation. Since CO is a component of syngas, a detailed understanding is crucial. In this study, a newly developed measurement setup is presented that allows sterile online measurement of the DCOT. In an abiotic experiment, the functionality of the measurement principle was demonstrated for various CO concentrations in the gas supply (0%-40%) and various agitation rates (300-1100 min-1 ). In continuous stirred tank reactor fermentation experiments, the measurement showed reliable results. The production of ethanol and 2,3-butanediol increased with increasing DCOT. Moreover, a critical DCOT was identified, leading to the inhibition of the culture. Thus, the reported online measurement method is beneficial for process understanding. In future processes, it can be used for closed-loop fermentation control.

Gas Fermentation Expands the Scope of a Process Network for Material Conversion

AbstractBiotechnological fermentation is a well‐established process, however, it is far from being fully understood and exploited. A new area of fermentation technology that has evolved over the recent decades is gas fermentation. Many microorganisms have been reported in literature to be capable of utilizing a variety of gases such as CO, CO2, H2, and CH4 under anaerobic or aerobic conditions as their main carbon and/or energy source. Mostly waste stream gases from industrial plants or those that can be produced via the gasification of solids are investigated. This review focuses on the currently available scientific knowledge about gas fermentation processes, particularly anaerobic syngas fermentation and aerobic methane fermentation. Gas fermentation processes are compared with aerobic and anaerobic fermentation processes based on dissolved solid substrates. Also, the potential of gas fermentation when integrated into a biotechnological network of processes is outlined.

Combination of Trace Metal to Improve Solventogenesis of Clostridium carboxidivorans P7 in Syngas Fermentation.

Higher alcohols such as butanol (C4 alcohol) and hexanol (C6 alcohol) are superior biofuels compared to ethanol. Clostridium carboxidivorans P7 is a typical acetogen capable of producing C4 and C6 alcohols natively. In this study, the composition of trace metals in culture medium was adjusted, and the effects of these adjustments on artificial syngas fermentation by C. carboxidivorans P7 were investigated. Nickel and ferrous ions were essential for growth and metabolite synthesis during syngas fermentation by P7. However, a decreased dose of molybdate improved alcohol fermentation performance by stimulating carbon fixation and solventogenesis. In response to the modified trace metal composition, cells grew to a maximum OD600nm of 1.6 and accumulated ethanol and butanol to maximum concentrations of 2.0 and 1.0 g/L, respectively, in serum bottles. These yields were ten-fold higher than the yields generated using the original composition of trace metals. Furthermore, 0.5 g/L of hexanol was detected at the end of fermentation. The results from gene expression experiments examining genes related to carbon fixation and organic acid and solvent synthesis pathways revealed a dramatic up-regulation of the Wood–Ljungdahl pathway (WLP) gene cluster, the bcs gene cluster, and a putative CoA transferase and butanol dehydrogenase, thereby indicating that both de novo synthesis and acid re-assimilation contributed to the significantly elevated accumulation of higher alcohols. The bdh35 gene was speculated to be the key target for butanol synthesis during solventogenesis.

Improved robustness of microbial electrosynthesis by adaptation of a strict anaerobic microbial catalyst to molecular oxygen

Microbial electrosynthesis (MES) and other bioprocesses such as syngas fermentation developed for energy storage and the conversion of carbon dioxide into valuable chemicals often employs acetogens as microbial catalysts. Acetogens are sensitive to molecular oxygen, which means that bioproduction reactors must be maintained under strict anaerobic conditions. This requirement increases cost and does not eliminate the possibility of O2 leakage. For MES, the risk is even greater since the system generates O2 when water splitting is the anodic reaction. Here, we show that O2 from the anode of a MES reactor diffuses into the cathode chamber where strict anaerobes reduce CO2. To overcome this drawback, a stepwise adaptive laboratory evolution (ALE) strategy is used to develop the O2 tolerance of the acetogen Sporomusa ovata. Two heavily-mutated S. ovata strains growing well autotrophically in the presence of 0.5 to 5% O2 were obtained. The adapted strains were more performant in the MES system than the wild type converting electrical energy and CO2 into acetate 1.5 fold faster. This study shows that the O2 tolerance of acetogens can be increased, which leads to improvement of the performance and robustness of energy-storage bioprocesses such as MES where O2 is an inhibitor.

Circumventing Unintended Impacts of Waste N95 Facemask Generated during the COVID-19 Pandemic: A Conceptual Design Approach

The global crisis arising from the current COVID-19 pandemic has resulted in a surge in the magnitude of global waste from used Personal Protective Equipment with special emphasis on waste N95 facemask. Creative approaches are therefore required to resolve the surging facemask waste disposal issue in an economical and environmentally friendly manner. In an attempt to resolve the evolving global waste challenge, the present study has assessed the economic and environmental performances of converting N95 facemasks to steam and electricity via a combined heat and power plant, to ethanol via a syngas fermentation process, and to an energy-dense gasoline-like oil product via a hydrothermal liquefaction process. These processes were assessed using “conceptual” process models developed using ASPEN plus as the process simulation tool. Economic and environment assessments were undertaken using net present values (NPVs) and the rate of potential environmental impacts (PEIs) respectively, as sufficient performance measures. Therefore, the present study was able to establish that the conversion of waste N95 facemask to syngas prior to a fermentation process for ethanol production constituted the least economical and least environmental friendly process with a negative NPV and the highest rate of PEI (1.59 PEI/h) value calculated. The NPV values calculated for N95 facemask waste conversion to steam and electricity and energy-dense oil processes were US$ 36.6 × 106 and US$ 53 × 106 respectively, suggesting the preference for the production of a valuable energy-dense oil product. Furthermore, it was observed that when the environmental performance of both processes was considered, rates of PEIs of 1.20 and 0.28 PEI/h were estimated for the energy-dense oil production process and the steam and electricity generation process, respectively. Therefore, the study was able to establish that the utilisation of waste N95 facemask for steam and electricity generation and for generating an energy-dense oil product are both promising approaches that could aid in the resolution of the waste issue if both environmental and economic performances constitute crucial considerations.

Batch Syngas Fermentation by Clostridium carboxidivorans for Production of Acids and Alcohols

Syngas (CO, CO2, and H2) has attracted special attention due to the double benefit of syngas fermentation for carbon sequestration (pollution reduction), while generating energy. Syngas can be either produced by gasification of biomasses or as a by-product of industrial processes. Only few microorganisms, mainly clostridia, were identified as capable of using syngas as a substrate to produce medium chain acids, or alcohols (such as butyric acid, butanol, hexanoic acid, and hexanol). Since CO plays a critical role in the availability of reducing equivalents and carbon conversion, this work assessed the effects of constant CO partial pressure (PCO), ranging from 0.5 to 2.5 atm, on cell growth, acid production, and solvent production, using Clostridium carboxidivorans. Moreover, this work focused on the effect of the liquid to gas volume ratio (VL/VG) on fermentation performances; in particular, two VL/VG were considered (0.28 and 0.92). The main results included—(a) PCO affected the growth kinetics of the microorganism; indeed, C. carboxidivorans growth rate was characterized by CO inhibition within the investigated range of CO concentration, and the optimal PCO was 1.1 atm (corresponding to a dissolved CO concentration of about 25 mg/L) for both VL/VG used; (b) growth differences were observed when the gas-to-liquid volume ratio changed; mass transport phenomena did not control the CO uptake for VL/VG = 0.28; on the contrary, the experimental CO depletion rate was about equal to the transport rate in the case of VL/VG = 0.92.

Immobilization of trophic anaerobic acetogen for semi-continuous syngas fermentation

Abstract The massive consumption of fossil energy forces people to find new sources of energy. Syngas fermentation has become a hot research field as its high potential in renewable energy production and sustainable development. In this study, trophic anaerobic acetogen Morella thermoacetica was successfully immobilized by calcium alginate embedding method. The ability of the immobilized cells on production of acetic acid through syngas fermentation was compared in both airlift and bubble column bioreactors. The bubble column bioreactor was selected as the better type of bioreactor. The production of acetic acid reached 32.3 g·L−1 in bubble column bioreactor with a space–time yield of 2.13 g·L−1·d−1. The immobilized acetogen could be efficiently reused without significant lag period, even if exposed to air for a short time. A semi-continuous syngas fermentation was performed using immobilized cells, with an average space–time acetic acid yield of 3.20 g·L−1·d−1. After 30 days of fermentation, no significant decrease of the acetic acid production rate was observed.

The effect of corn syrup and whey on the conversion process of CO to ethanol using Clostridium ljungdahlii

CO is one of the toxic components of syngas, which is the major source of air pollution. Syngas fermentation technology has the ability to convert toxic gases into valuable biofuels, such as ethanol. Fermentative ethanol production is an important method that can be used to promote environmental protection. CO can be converted into ethanol, via the Wood–Ljungdahl pathway, using Clostridium ljungdahlii. The components of the growing medium––especially the trace-element solution and yeast extract––are the main reasons for the high costs associated with this process, however, and this especially impacts scaled-up operations. In this study, cheaper substitutes for these components were used in order to determine their effect on ethanol production. The study comprised three main parts––the optimization of CO concentration, and the substitution of corn syrup and whey powder in the process. The optimum volume of CO for ethanol production was found to be 10 mL. Corn syrup can be used instead of trace-element solution, but the use of yeast extract with the corn syrup was determined to be essential. Up to 1.4 g/L ethanol production was observed with the addition of 15 mL corn syrup. Whey powder had the advantage of being usable without yeast extract, with up to 2.5 g/L ethanol being produced from a 30-g/L concentration. The main finding was that either corn syrup or whey powder can be used as substitutes for expensive basal-medium components.

Simulation and Performance Analysis of Integrated Gasification–Syngas Fermentation Plant for Lignocellulosic Ethanol Production

This study presents a new simulation model developed with ASPEN Plus of waste biomass gasification integrated with syngas fermentation and product recovery units for bioethanol production from garden waste as a lignocellulosic biomass. The simulation model includes three modules: gasification, fermentation, and ethanol recovery. A parametric analysis is carried out to investigate the effect of gasification temperature (500–1500 °C) and equivalence ratio (0.2–0.6) on the gasification performance and bioethanol production yield. The results reveal that, for efficient gasification and high ethanol production, the operating temperature range should be 700–1000 °C, as well as an equivalence ratio between 0.2 and 0.4. At optimal operating conditions, the bioethanol production yield is 0.114 kg/h per 1 kg/h input garden waste with 50% moisture content. It is worth mentioning that this parameter increases to 0.217 kgbioethanol/kggarden waste under dry-based conditions.

Evaluation of Media Components and Process Parameters in a Sensitive and Robust Fed-Batch Syngas Fermentation System with Clostridium ljungdahlii

The fermentation of synthesis gas, or syngas, by acetogenic bacteria can help in transitioning from a fossil-fuel-based to a renewable bioeconomy. The main fermentation products of Clostridium ljungdahlii, one of such microorganisms, are acetate and ethanol. A sensitive, robust and reproducible system was established for C. ljungdahlii syngas fermentation, and several process parameters and medium components (pH, gas flow, cysteine and yeast extract) were investigated to assess its impact on the fermentation outcomes, as well as real time gas consumption. Moreover, a closed carbon balance could be achieved with the data obtained. This system is a valuable tool to detect changes in the behavior of the culture. It can be applied for the screening of strains, gas compositions or media components, for a better understanding of the physiology and metabolic regulation of acetogenic bacteria. Here, it was shown that neither yeast extract nor cysteine was a limiting factor for cell growth since their supplementation did not have a noticeable impact on product formation or overall gas consumption. By combining the lowering of both the pH and the gas flow after 24 h, the highest ethanol to acetate ratio was achieved, but with the caveat of lower productivity.

Medium Compositions for the Improvement of Productivity in Syngas Fermentation with Clostridium autoethanogenum

The cost of culture media represents a considerable portion of the overall cost in syngas fermentation. Yeast extract is an expensive ingredient among the medium components required to prepare the culture medium. In this study, we evaluated the effectiveness of replacing yeast extract with relatively low-cost nutrient supplements such as corn steep liquor, malt extract, and vegetable extract in improving the feasibility of the syngas fermentation process in Clostridium autoethanogenum culture. The use of corn steep liquor, malt extract, and vegetable extract as nutrient supplements at a concentration of 0.5 g/L resulted in optical densities of 1.44, 1.37, and 1.71, and produced ethanol concentrations of 2.24, 3.37, and 3.76 g/L, respectively. The microbial growth and ethanol production in syngas fermentation with yeast extract alternatives was comparable with the results of fermentation with yeast extract (optical density of 1.48 and ethanol concentration of 2.16). Moreover, the use of malt and vegetable extracts showed considerable improvement (69% and 51%, respectively) in the specific ethanol productivity compared to yeast extract.

Behavior of CO-water mass transfer coefficient in membrane sparger-integrated bubble column for synthesis gas fermentation

The gas–liquid mass transfer coefficient (kLa) of O2 was investigated in a bubble column reactor (BCR) using a sintered gas filter (SF), ceramic membrane module (CMM), and hollow fiber membrane module (HFM), which have different ranges of gas supply areas. kLa was enhanced by increasing flow rate in all of the spargers. Different responses when changing the gas supply area were obtained depending on the sparger type. Average values of kLa that were 52 and 258% higher were obtained using a CMM-integrated BCR compared to SFs and HFMs. CO-water kLa was investigated using CMMs for application to gas fermentation. The CO-water kLa ranged from 28.3 to 113.7/h under the experimental conditions. Based on the experimental data from CO and O2, a model to predict kLa was constructed for CMM-integrated BCRs. A dimensionless number indicating a gas supply area of the sparger was newly defined and included in the developed model.

Bioethanol production through syngas fermentation in a tar free bioreactor using Clostridium butyricum

Biomass-generated syngas conversion through fermentation is a promising technique for bioethanol production due to increasing demand for clean and sustainable energy. However, lignocellulosic biomass is difficult to degrade wholly, and traditional pretreatment process has several drawbacks. The present study emphasizes on bioethanol production from lignocellulosic biomass-based syngas including the main composition of N2 (45.58%), CO (22.92%), CO2 (7.9%), H2 (13.05%), and CH4 (1.13%). Field emission electronic microscopic analysis was used to characterize freshly cultured Clostridium butyricum for syngas fermentation and experiment was run in a bioreactor (TFB). The obtained yield of bioethanol was analyzed by nuclear magnetic resonance and gas chromatography-mass spectrometry analyses. For this syngas fermentation, treated syngas was preferred, as most of the Clostridium butyricum grown on best fermentation conditions. The results show that except 0.03% of CO2, other gases were dissolved entirely. It is also found that extracted bioethanol was identified by corresponding NMR (1H) spectra of methyle group (CH3-), methylene group (–CH2–) and hydroxyl group (OH). The yield of bioethanol was 29.94 mmol from 1 L of syngas. Hence, this biomass-generated syngas is the appropriate renewable energy source for the meetup of future energy needs.

Production routes of advanced renewable C1 to C4 alcohols as biofuel components – a review

AbstractIn 2018, the EU's revised Renewable Energy Directive came into force, increasing renewable energy targets for all energy sectors while limiting the use of first‐generation biomass as feedstock. Fuel components like methanol, ethanol, propanols, and butanols represent promising candidates to enable the targets for transportation to be achieved because they can be used with existing infrastructure and can be further processed to give additives and substitutes. A wide variety of feedstocks and processes are available for this purpose. In this review, the thermocatalytic and biological synthesis routes for C1–C4 alcohol fuels are summarized to illustrate the many alternatives. They include biomass and waste gasification and carbon capture and utilization to obtain syngas for catalytic conversion, fermentation of sugars from lignocellulosic feedstock, and novel, less developed pathways like syngas fermentation, glycerol conversion, and biogas reforming. The current state of technology is presented by discussing the advantages and technical hurdles, and by introducing recent scaled‐up approaches. This demonstrates the need for further research and development. The assessment of techno‐economic analyses in the literature illustrates the dominant factors affecting production costs and reveals the broad range of feasibility of the various production routes. The review shows that the routes most similar to conventional, well‐established syntheses bear the highest potential to be implemented in the short and medium term. The availability of cheap and abundant feedstock also plays a crucial role. Methanol synthesis from biomass gasification and ethanol, and acetone‐butanol‐ethanol fermentation from lignocellulosic biomass, are therefore considered to be very promising. © 2020 The Authors. Biofuels, Bioproducts and Biorefining published by Society of Industrial Chemistry and John Wiley & Sons Ltd.