Search

Background: Ethyl acetate is a bulk chemical traditionally produced via energy intensive chemical esterification. Microbial production of this compound offers promise as a more sustainable alternative process. So far, efforts have focused on using sugar-based feedstocks for microbial ester production, but extension to one-carbon substrates, such as CO and CO 2 /H 2 , is desirable. Acetogens present a promising microbial platform for the production of ethyl esters from these one-carbon substrates., Results: We engineered the acetogen C. autoethanogenum to produce ethyl acetate from CO by heterologous expression of an alcohol acetyltransferase (AAT), which catalyzes the formation of ethyl acetate from acetyl-CoA and ethanol. Two AATs, Eat1 from Kluyveromyces marxianus and Atf1 from Saccharomyces cerevisiae, were expressed in C. autoethanogenum. Strains expressing Atf1 produced up to 0.2 mM ethyl acetate. Ethyl acetate production was barely detectable (< 0.01 mM) for strains expressing Eat1. Supplementation of ethanol was investigated as potential boost for ethyl acetate production but resulted only in a 1.5-fold increase (0.3 mM ethyl acetate). Besides ethyl acetate, C. autoethanogenum expressing Atf1 could produce 4.5 mM of butyl acetate when 20 mM butanol was supplemented to the growth medium., Conclusions: This work offers for the first time a proof-of-principle that autotrophic short chain ester production from C1-carbon feedstocks is possible and offers leads on how this approach can be optimized in the future., (© 2022. The Author(s).)

Biochar feedstock and production method affects its physicochemical properties and subsequent application. This study investigated the effects of biochar from switchgrass (SGB) and poultry litter (PLB) produced at 350 and 700 °C on alcohol formation using CO:CO 2 :H 2 (40:30:30) by Clostridium carboxidivorans (P7) and C. ragsdalei (P11). Fermentations were performed in 250 mL bottles with a 50 mL working volume at 37 °C. Strains P7 and P11 produced 1.2- to 2.2-fold more alcohol and consumed 1.2- to 1.9-fold more syngas using biochars made at 700 °C compared to 350 °C. Both strains also produced 1.4- to1.9-fold more alcohol with both biochars made at 700 °C compared to control without biochar. Strain P11 produced 1.1- and 1.6-fold more alcohol and fatty acids, respectively, in medium with PLB made at 700 °C compared to strain P7. These results provide guidance towards the selection of biochar type and production temperature to improve syngas fermentation., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Syngas fermentation has been successfully implemented in commercial-scale plants and can enable the biochemical conversion of the driest fractions of biomass through synthesis gas (H 2 , CO 2 , and CO). The process relies on optimized acetogenic strains able to reach and maintain high productivity of ethanol and acetate. In parallel, microbial communities have shown to be the best choice for the production of valuable medium-chain carboxylates through anaerobic fermentation of biomass, demanding low technical complexity and being able to realize simultaneous hydrolysis of the substrate. Each of the two technologies benefits from different strong points and has different challenges to overcome. This review discusses the rationales for merging these two seemingly disparate technologies by analyzing previous studies and drawing opinions based on the lessons learned from such studies. For keeping the technical demands of the resulting process low, a case is built for using microbial communities instead of pure strains. For that to occur, a shift from conventional syngas-based to "syngas-aided" anaerobic fermentation is suggested. Strategies for tackling the intricacies of working simultaneously with communities and syngas, such as competing pathways, and thermodynamic aspects are discussed as well as the stoichiometry and economic feasibility of the concept. Overall, syngas-aided anaerobic fermentation seems to be a promising concept for the biorefinery of the future. However, the effects of process parameters on microbial interactions have to be understood in greater detail, in order to achieve and sustain feasible medium-chain carboxylate and alcohol productivity.

Clostridium carboxidivorans can convert CO 2 , CO and H 2 to ethanol and n-butanol; however, its industrial application is limited by the lack of tools for metabolic pathway engineering. In this study, C. carboxidivorans was successfully engineered to overexpress aor, adhE2, and fnr together with adhE2 or aor. In glucose fermentation, all engineered strains showed higher alcohol yields compared to the wild-type. Strains overexpressing aor showed CO 2 re-assimilation during heterotrophic growth. In syngas fermentation, compared to the wild-type, the strain overexpressing adhE2 produced ∼50% more ethanol and the strain overexpressing adhE2 and fnr produced ∼18% more butanol and ∼22% more ethanol. Interestingly, both strains showed obvious acid re-assimilation, with <0.15 g/L total acid remaining at the end of fermentation. Overexpressing fnr with adhE2 enhanced butanol production compared to only adhE2. This is the first report of overexpressing homologous and heterologous genes in C. carboxidivorans for enhancing alcohols production from syngas and glucose., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Biochar has functional groups, pH buffering capacity and cation exchange capacity (CEC) that can be beneficial in syngas fermentation. This study examined the properties of biochar made from switchgrass (SGBC), forage sorghum (FSBC), red cedar (RCBC) and poultry litter (PLBC), and their effects on ethanol and butanol production from syngas using Clostridium carboxidivorans. Experiments were performed in 250 mL bottle reactors with a 50 mL working volume at 37 °C fed syngas containing CO:H 2 :CO 2 (40:30:30 by volume). Results showed that PLBC and SGBC enhanced ethanol production by 90% and 73%, respectively, and butanol production by fourfold compared to standard yeast extract medium without biochar (control). CO and H 2 utilization in PLBC and SGBC media increased compared to control. PLBC had the highest pH buffering capacity, CEC and total amount of cations compared with SGBC, FSBC and RCBC, which could have contributed to its highest enhancement of ethanol and butanol production., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This study investigated the effects of electrolytes (CaCl 2 , K 2 HPO 4 , MgSO 4 , NaCl, and NH 4 Cl) on CO mass transfer and ethanol production in a HFMBR. The hollow fiber membranes (HFM) were found to generate tiny gas bubbles; the bubble coalescence was significantly suppressed in electrolyte solution. The volumetric gas-liquid mass transfer coefficients (k L a) increased up to 414% compared to the control. Saturated CO (C ∗ ) decreased as electrolyte concentrations increased. Overall, the maximum mass transfer rate (R max ) in electrolyte solution ranged from 106% to 339% of the value obtained in water. The electrolyte toxicity on cell growth was tested using Clostridium autoethanogenum. Most electrolytes, except for MgSO 4 , inhibited cell growth. The HFMBR operation using a medium containing 1% MgSO 4 achieved 119% ethanol production compared to that without electrolytes. Finally, a kinetic simulation using the parameters got from the 1% MgSO 4 medium predicted a higher ethanol production compared to the control., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Renewable energy, including biofuels such as ethanol and butanol from syngas bioconversed by Clostridium carboxidivorans P7, has been drawing extensive attention due to the fossil energy depletion and global eco-environmental issues. Effects of zinc on the growth and metabolites of C. carboxidivorans P7 were investigated with model syngas as the carbon source. The cell concentration was doubled, the ethanol content increased 3.02-fold and the butanol content increased 7.60-fold, the hexanol content increased 44.00-fold in the medium with 280 μM Zn 2+ , when comparing with those in the control medium [Zn 2+ , (7 μM)]. Studies of the genes expression involved in the carbon fixation as well as acid and alcohol production in the medium with 280 μM Zn 2+ indicated that fdhII was up-regulated on the second day, acs A, fdhII, bdh35 and bdh50 were up-regulated on the third day and bdh35, acsB, fdhI, fdhIII, fdhIV, buk, bdh10, bdh35, bdh40 and bdh50 were up-regulated on the fourth day. The results indicated that the increased Zn 2+ content increased the alcohol production through increase in the gene expression of the carbon fixation and alcohol dehydrogenase.

Biochar contains minerals and metals that can serve as nutrients for acetogens to produce ethanol via syngas fermentation. In this study, four fermentation media containing biochar from switchgrass (SGBC), forage sorghum (FSBC), red cedar (RCBC) and poultry litter (PLBC) were compared with standard yeast extract (YE) medium for syngas fermentation using Clostridium ragsdalei. Fermentations were performed in 250mL bottle reactors at 150rpm and 37°C with syngas containing CO:H 2 :CO 2 (40:30:30) by volume. Results showed that media containing RCBC and PLBC improved ethanol production by 16.3% and 58.9%, respectively, compared to YE medium. C. ragsdalei consumed 69% more H 2 and 40% more CO in PLBC medium compared to YE medium. However, no enhancement of ethanol production was observed in SGBC and FSBC media. The highest release of Na, K, Ca, Mg, S and P was from PLBC, which was considered to contribute in enhancement of ethanol production., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Syngas fermentation is an anaerobic bioprocess that could become industrially relevant as a biorefinery platform for sustainable production of fuels and chemicals. An important prerequisite for commercialization is adequate performance of the biocatalyst (i.e., sufficiently high production rate, titer, selectivity, yield, and stability of the fermentation). Here, we compared the performance of three potential candidate Clostridium strains in syngas-to-ethanol conversion: Clostridium ljungdahlii PETC, C. ljungdahlii ERI-2, and Clostridium autoethanogenum JA1-1. Experiments were conducted in a two-stage, continuously fed syngas-fermentation system that had been optimized for stable ethanol production. The two C. ljungdahlii strains performed similar to each other but different from C. autoethanogenum. When the pH value was lowered from 5.5 to 4.5 to induce solventogenesis, the cell-specific carbon monoxide and hydrogen consumption (similar rate for all strains at pH 5.5), severely decreased in JA1-1, but hardly in PETC and ERI-2. Ethanol production in strains PETC and ERI-2 remained relatively stable while the rate of acetate production decreased, resulting in a high ethanol/acetate ratio, but lower overall productivities. With JA1-1, lowering the pH severely lowered rates of both ethanol and acetate production; and as a consequence, no pronounced shift to solventogenesis was observed. The highest overall ethanol production rate of 0.301 g · L(-1)  · h(-1) was achieved with PETC at pH 4.5 with a corresponding 19 g/L (1.9% w/v) ethanol concentration and a 5.5:1 ethanol/acetate molar ratio. A comparison of the genes relevant for ethanol metabolism revealed differences between C. ljungdahlii and C. autoethanogenum that, however, did not conclusively explain the different phenotypes., (© 2015 Wiley Periodicals, Inc.)

Incubation of carboxydotrophs at 37 °C provides optimal conditions for their growth. However, a fast accumulation of organic acids, specifically acetate, during the exponential growth phase may result in low alcohol production and substrate consumption due to a phenomenon known as "acid crash". The present work investigates growth and productivity of Clostridium carboxidivorans P7 at two incubation temperatures. At 37 °C the culture was not able to override the "acid crash", resulting in low ethanol titers (1.56 mM). On the other hand, lower metabolic rates at 25 °C enhanced ethanol and butanol production (32.1 and 14.5 mM, respectively). Moreover, at low temperatures, hexanol and caproic acid were also produced at significant concentrations, 8.21 and 9.02 mM respectively, among the highest values reported for P7. Our results demonstrate that production of biofuels with longer carbon chains molecules may be enhanced incubating syngas-fermenting acetogenic bacteria at sub-optimal temperatures., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The current energy model based on fossil fuels is coming to an end due to the increase in global energy demand. Biofuels such as ethanol and butanol can be produced through the syngas fermentation by acetogenic bacteria. The present work hypothesizes that formate addition would positively impact kinetic parameters for growth and alcohol production in Clostridium ljungdahlii PETC and Clostridium carboxidivorans P7 by diminishing the need for reducing equivalents. Fermentation experiments were conducted using completely anaerobic batch cultures at different pH values and formate concentrations. PETC cultures were more tolerant to formate concentrations than P7, specially at pH 5.0 and 6.0. Complete growth inhibition of PETC occurred at sodium formate concentrations of 30.0 mM; however, no differences in growth rates were observed at pH 7.0 for the two strains. Incubation at formate concentrations lower than 2.0 mM resulted in increased growth rates for both strains. The most recognizable effects of formate addition on the fermentation products were the increase in the total carbon fixed into acids and alcohols at pH 5.0 and pH 6.0, as well as, a higher ethanol to total products ratio at pH 7.0. Taken all together, these results show the ability of acetogens to use formate diminishing the energy demand for growth, and enhancing strain productivity., (Copyright© by the Spanish Society for Microbiology and Institute for Catalan Studies.)

The development of a low cost medium for ethanol production is critical for process feasibility. Ten media were formulated for Clostridium ragsdalei by reduction, elimination and replacement of expensive nutrients. Cost analysis and effects of medium components on growth and product formation were investigated. Fermentations were performed in 250 mL bottles using syngas (20% CO, 15% CO2, 5% H2 and 60% N2). The standard medium M1 cost is $9.83/L, of which 93% is attributed to morpholinoethane sulfonic acid (MES) buffer. Statistical analysis of the results showed that MES removal did not affect cell growth and ethanol production (P>0.05). Based on cells' elemental composition, a minimal mineral concentration medium M7 was formulated, which provided 29% higher ethanol yield from CO at 3% of the cost compared to medium M1. Ethanol yield from CO in the completely defined medium M9 was 36% higher than while at 5% the cost of medium M1., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The use of mixed cultures in gas fermentations could reduce operating costs in the production of liquid chemicals such as alcohols or carboxylic acids. However, directing reducing equivalents towards the desired products presents the challenge of co-existing competing pathways. In this study, two trickle bed reactors were operated at acetogenic and chain elongating conditions to explore the fate of electron equivalents (ethanol, H2, and CO) and test pH oscillations as a strategy to target chain-elongated products. Hereby, the use of a H2-rich syngas increased gas conversion rates and the specificity towards acetic acid (86% of C-mol production, 9.0 g LEBV−1 day−1, with EBV referring to empty bed volume), while preliminary experiments with CO-rich syngas show promising results in increasing the ethanol production necessary to target chain-elongated products. On the other hand, ethanol supplementation hindered the endogenous ethanol production of the acetogenic culture but promoted butanol production (1.0 g LEBV−1 day−1) at high ethanol concentrations (9.6 g L−1) in the fresh media. Finally, pH oscillations improved chain elongation yields but negatively affected acetogenic growth, reducing production rates. [ABSTRACT FROM AUTHOR]

Syngas fermentation with clostridial co-cultures is promising for the conversion of CO to alcohols. A CO sensitivity study with Clostridium kluyveri monocultures in batch operated stirred-tank bioreactors revealed total growth inhibition of C. kluyveri already at 100 mbar CO, but stable biomass concentrations and ongoing chain elongation at 800 mbar CO. On/off-gassing with CO indicated a reversible inhibition of C. kluyveri. A continuous supply of sulfide led to increased autotrophic growth and ethanol formation by Clostridium carboxidivorans even at unfavorable low CO concentrations. Based on these results, a continuously operated cascade of two stirred-tank reactors was established with a synthetic co-culture of both Clostridia. An amount of 100 mbar CO and additional sulfide supply enabled growth and chain elongation in the first bioreactor, whereas 800 mbar CO resulted in an efficient reduction of organic acids and de-novo synthesis of C2-C6 alcohols in the second reactor. High alcohol/acid ratios of 4.5–9.1 (w/w) were achieved in the steady state of the cascade process, and the space-time yields of the alcohols produced were improved by factors of 1.9–5.3 compared to a batch process. Further improvement of continuous production of medium chain alcohols from CO may be possible by applying less CO-sensitive chain-elongating bacteria in co-cultures. [ABSTRACT FROM AUTHOR]

The effect of tungsten and selenium on cell growth and production of metabolites such as acetic acid and ethanol when fermenting syngas using " Clostridium autoethanogenum" was investigated to improve the process efficiency. General concentrations of selenium and tungsten in the medium are 0.01 µM during acetogenic syngas fermentation. We conducted culture experiments at concentrations of 0, 0.001, 0.01 and 0.1 µM for each heavy metal. The effect of selenium on cell growth and total metabolite production was greater than that of tungsten as the effect of selenium on formate dehydrogenase, an important enzyme of the Wood-Ljungdahl pathway, is greater than that of tungsten. Although an increase in tungsten had a marginal effect on total metabolite production, the ethanol/acetic acid production ratio increased significantly due to a decrease in acetic acid and an increase in ethanol production. Thus, tungsten plays a key role in activating aldehyde:ferredoxin oxidoreductase, a key enzyme in the reduction of acetate to ethanol. A specific ethanol productivity of 0.462 g ethanol/g DCW∙d was obtained in a culture using 0.01 µM selenium and 0.1 µM tungsten, which was 2.18 times higher than when using 0.01 µM of both selenium and tungsten. • Effect of selenium and tungsten on syngas fermentation was investigated. • Selenium showed greater effect in cell growth and C2 compounds production. • High tungsten dosage increased ethanol/acetic acid production ratio. • High tungsten concentration in medium significantly enhanced ethanol production. [ABSTRACT FROM AUTHOR]

Abstract Background Ethanol production through fermentation of gas mixtures containing CO, CO2 and H2 has just started operating at commercial scale. However, quantitative schemes for understanding and predicting productivities, yields, mass transfer rates, gas flow profiles and detailed energy requirements have been lacking in literature; such are invaluable tools for process improvements and better systems design. The present study describes the construction of a hybrid model for simulating ethanol production inside a 700 m3 bubble column bioreactor fed with gas of two possible compositions, i.e., pure CO and a 3:1 mixture of H2 and CO2. Results Estimations made using the thermodynamics-based black-box model of microbial reactions on substrate threshold concentrations, biomass yields, as well as CO and H2 maximum specific uptake rates agreed reasonably well with data and observations reported in literature. According to the bioreactor simulation, there is a strong dependency of process performance on mass transfer rates. When mass transfer coefficients were estimated using a model developed from oxygen transfer to water, ethanol productivity reached 5.1 g L−1 h−1; when the H2/CO2 mixture is fed to the bioreactor, productivity of CO fermentation was 19% lower. Gas utilization reached 23 and 17% for H2/CO2 and CO fermentations, respectively. If mass transfer coefficients were 100% higher than those estimated, ethanol productivity and gas utilization may reach 9.4 g L−1 h−1 and 38% when feeding the H2/CO2 mixture at the same process conditions. The largest energetic requirements for a complete manufacturing plant were identified for gas compression and ethanol distillation, being higher for CO fermentation due to the production of CO2. Conclusions The thermodynamics-based black-box model of microbial reactions may be used to quantitatively assess and consolidate the diversity of reported data on CO, CO2 and H2 threshold concentrations, biomass yields, maximum substrate uptake rates, and half-saturation constants for CO and H2 for syngas fermentations by acetogenic bacteria. The maximization of ethanol productivity in the bioreactor may come with a cost: low gas utilization. Exploiting the model flexibility, multi-objective optimizations of bioreactor performance might reveal how process conditions and configurations could be adjusted to guide further process development.

Syngas fermentation allows for the conversion of wastes into useful commodity chemicals. To target higher value products, the conditions can be tuned to be favourable for both acetogenic and reverse beta-oxidation pathways and produce, in one stage, butyric and caproic acid. Studies in CSTR have shown the crucial role of pH, which must be low enough to allow for ethanol generation in the acetogenic step while avoiding the inhibition of reverse β-oxidation in acidic conditions. However, no studies have investigated the effect of pH in reactor configurations suitable for syngas fermentation (i.e., allowing for cell retention and exhibiting high mass transfer rates at low operating costs), such as Trickle Bed Reactors, TBR. In this study, two TBR were used to study the pH effect on the fermentation of syngas to produce C4 and C6 acids, using undefined mixed cultures. Five pH values were tested in the range 4.5–7.5, and pH 6 was found to be the most favourable for simultaneous production of C4 & C6 acids from syngas, which agrees with what was found in suspended growth systems. In addition, the highest titers in literature so far were achieved in the TRB. 16S rRNA analysis was performed showing Clostridium and Rummenliibacillus to be the key genus for the efficient process at pH 6. Finally, the experimental methodology followed, and data collected proved the robustness of mixed culture biofilm reactors in respect to pH changes, as the same reactor performance and bacterial community were achieved regardless of the operation history. [Display omitted] • High titers, productivities, and yields of C4 & C6 acids were obtained. • pH 6 was most favourable for chain elongation in trickle bed reactors. • Microbial community and performance were not affected by the operation history. • Clostridium and Rummeliibacillus dominated the community at pH 6. [ABSTRACT FROM AUTHOR]

Abstract Background Gas-fermenting acetogens have received a great deal of attention for their ability to grow on various syngas and waste gas containing carbon monoxide (CO), producing acetate as the primary metabolite. Among them, some Clostridium species, such as C. ljungdahlii and C. autoethanogenum, are of particular interest as they produce fuel alcohols as well. Despite recent efforts, alcohol production by these species is still unsatisfactory due to their low productivity and acetate accumulation, necessitating the isolation of strains with better phenotypes. Results In this study, a novel alcohol-producing acetogen (Clostridium sp. AWRP) was isolated, and its complete genome was sequenced. This bacterium belongs the same phylogenetic group as C. ljungdahlii, C. autoethanogenum, C. ragsdalei, and C. coskatii based on 16S rRNA homology; however, the levels of genome-wide average nucleotide identity (gANI) for strain AWRP compared with these strains range between 95 and 96%, suggesting that this strain can be classified as a novel species. In addition, strain AWRP produced a substantial amount of ethanol (70–90 mM) from syngas in batch serum bottle cultures, which was comparable to or even exceeded the typical values obtained using its close relatives cultivated under similar conditions. In a batch bioreactor, strain AWRP produced 119 and 12 mM of ethanol and 2,3-butanediol, respectively, while yielding only 1.4 mM of residual acetate. Interestingly, the alcohologenesis of this strain was strongly affected by oxidoreduction potential (ORP), which has not been reported with other gas-fermenting clostridia. Conclusion Considering its ethanol production under low oxidoreduction potential (ORP) conditions, Clostridium sp. AWRP will be an interesting host for biochemical studies to understand the physiology of alcohol-producing acetogens, which will contribute to metabolic engineering of those strains for the production of alcohols and other value-added compounds from syngas.

Syngas, the product of biomass gasification, can play an important role in moving towards the production of renewable chemical commodities, by using acetogenic bacteria to ferment those gaseous mixtures. Due to the complex and changing nature of biomass, the composition and the impurities present in the final biomass-derived syngas will vary. Because of this, it is important to assess the impact of these factors on the fermentation outcome, in terms of yields, productivity, and product formation and ratio. In this study, Clostridium ljungdahlii was used in a fed-batch fermentation system to analyze the effect of three different biomass-derived syngases, and to compare them to equivalent, clean syngas mixtures. Additionally, four other clean syngas mixtures were used, and the effects on product ratio, productivity, yield, and growth were documented. All biomass-derived syngases were suitable to be used as substrates, without experiencing any complete inhibitory effects. From the obtained results, it is clear that the type of syngas, biomass-derived or clean, had the greatest impact on product formation ratios, with all biomass-derived syngases producing more ethanol, albeit with lesser total productivity. [ABSTRACT FROM AUTHOR]

Background: Ethanol production through fermentation of gas mixtures containing CO, CO2 and H2 has just started operating at commercial scale. However, quantitative schemes for understanding and predicting productivities, yields, mass transfer rates, gas flow profiles and detailed energy requirements have been lacking in literature; such are invaluable tools for process improvements and better systems design. The present study describes the construction of a hybrid model for simulating ethanol production inside a 700 m3 bubble column bioreactor fed with gas of two possible compositions, i.e., pure CO and a 3:1 mixture of H2 and CO2. Results: Estimations made using the thermodynamics-based black-box model of microbial reactions on substrate threshold concentrations, biomass yields, as well as CO and H2 maximum specific uptake rates agreed reasonably well with data and observations reported in literature. According to the bioreactor simulation, there is a strong dependency of process performance on mass transfer rates. When mass transfer coefficients were estimated using a model developed from oxygen transfer to water, ethanol productivity reached 5.1 g L−1 h−1; when the H2/CO2 mixture is fed to the bioreactor, productivity of CO fermentation was 19% lower. Gas utilization reached 23 and 17% for H2/CO2 and CO fermentations, respectively. If mass transfer coefficients were 100% higher than those estimated, ethanol productivity and gas utilization may reach 9.4 g L−1 h−1 and 38% when feeding the H2/CO2 mixture at the same process conditions. The largest energetic requirements for a complete manufacturing plant were identified for gas compression and ethanol distillation, being higher for CO fermentation due to the production of CO2. Conclusions: The thermodynamics-based black-box model of microbial reactions may be used to quantitatively assess and consolidate the diversity of reported data on CO, CO2 and H2 threshold concentrations, biomass yields, maximum substrate uptake rates, and half-saturation constants for CO and H2 for syngas fermentations by acetogenic bacteria. The maximization of ethanol productivity in the bioreactor may come with a cost: low gas utilization. Exploiting the model flexibility, multi-objective optimizations of bioreactor performance might reveal how process conditions and configurations could be adjusted to guide further process development. [ABSTRACT FROM AUTHOR]

Syngas fermentation is one of the bets for the future sustainable biobased economies due to its potential as an intermediate step in the conversion of waste carbon to ethanol fuel and other chemicals. Integrated with gasification and suitable downstream processing, it may constitute an efficient and competitive route for the valorization of various waste materials, especially if systems engineering principles are employed targeting process optimization. In this study, a dynamic multi‐response model is presented for syngas fermentation with acetogenic bacteria in a continuous stirred‐tank reactor, accounting for gas–liquid mass transfer, substrate (CO, H2) uptake, biomass growth and death, acetic acid reassimilation, and product selectivity. The unknown parameters were estimated from literature data using the maximum likelihood principle with a multi‐response nonlinear modeling framework and metaheuristic optimization, and model adequacy was verified with statistical analysis via generation of confidence intervals as well as parameter significance tests. The model was then used to study the effects of process conditions (gas composition, dilution rate, gas flow rates, and cell recycle) as well as the sensitivity of kinetic parameters, and multiobjective genetic algorithm was used to maximize ethanol productivity and CO conversion. It was observed that these two objectives were clearly conflicting when CO‐rich gas was used, but increasing the content of H2 favored higher productivities while maintaining 100% CO conversion. The maximum productivity predicted with full conversion was 2 g·L−1·hr−1 with a feed gas composition of 54% CO and 46% H2 and a dilution rate of 0.06 hr−1 with roughly 90% of cell recycle. [ABSTRACT FROM AUTHOR]

The production of ethanol via syngas fermentation obtained from lignocellulose gasification provides a method for completely utilizing all of the carbon content from lignocellulosic feedstock. The low mass transfer rate of less soluble CO and H2 gas to liquid has been considered a major bottleneck in the overall process; however, microporous membrane has been proposed as a gas diffuser to improve gas-to-liquid mass transfer. In this study, a liquid batch of syngas fermentation employing Clostridium ljungdahlii with continuous gas supply was obtained using the configuration of a bioreactor connected to microporous hydrophobic polypropylene hollow fiber membrane (HFM) as a gas diffuser. Liquid recirculation between the fermentation vessel and membrane module was applied to enhance the gas-liquid contact as well as cell-recycle. The fermentation performance with and without HFM was compared and evaluated by cell growth, CO utilization, ethanol yield, and productivity. A higher ethanol yield, 0.22 mol/mol, was achieved using the system of an HFM-supported bioreactor with a higher ethanol titer of 1.09 g/L and an ethanol-acetate molar ratio of 1.43 mol/mol. The obtained result demonstrates that an HFM-supported bioreactor is the best fermentation system compared to stirred tank reactor (STR) without a membrane. [ABSTRACT FROM AUTHOR]

The production of ethanol via syngas fermentation obtained from lignocellulose gasification provides a method for completely utilizing all of the carbon content from lignocellulosic feedstock. The low mass transfer rate of less soluble CO and H2 gas to liquid has been considered a major bottleneck in the overall process; however, microporous membrane has been proposed as a gas diffuser to improve gas-to-liquid mass transfer. In this study, a liquid batch of syngas fermentation employing Clostridium ljungdahlii with continuous gas supply was obtained using the configuration of a bioreactor connected to microporous hydrophobic polypropylene hollow fiber membrane (HFM) as a gas diffuser. Liquid recirculation between the fermentation vessel and membrane module was applied to enhance the gas–liquid contact as well as cell-recycle. The fermentation performance with and without HFM was compared and evaluated by cell growth, CO utilization, ethanol yield, and productivity. A higher ethanol yield, 0.22 mol/mol, was achieved using the system of an HFM-supported bioreactor with a higher ethanol titer of 1.09 g/L and an ethanol-acetate molar ratio of 1.43 mol/mol. The obtained result demonstrates that an HFM-supported bioreactor is the best fermentation system compared to stirred tank reactor (STR) without a membrane. [ABSTRACT FROM AUTHOR]

The hybrid gasification-syngas fermentation platform can produce more bioethanol utilizing all biomass components compared to the biochemical conversion technology. Syngas fermentation operates at mild temperatures and pressures and avoids using expensive pretreatment processes and enzymes. This study presents a new process simulation model developed with Aspen Plus® of a biorefinery based on a hybrid conversion technology for the production of anhydrous ethanol using 1200 tons per day (wb) of switchgrass. The simulation model consists of three modules: gasification, fermentation, and product recovery. The results revealed a potential production of about 36.5 million gallons of anhydrous ethanol per year. Sensitivity analyses were also performed to investigate the effects of gasification and fermentation parameters that are keys for the development of an efficient process in terms of energy conservation and ethanol production. [ABSTRACT FROM AUTHOR]

Fuel ethanol has been recognized as a kind of potential alternative fuel as well as an additive to gasoline because of its oxygenated, and high octane characteristics. Syngas fermentation, a novel route for ethanol production, is getting more and more attention. Syngas can be generated from a lot of organic materials including biomass. Gasification technology is used to convert the biomass into a mixture of gases (consisting mainly of CO, CO2 and H2), which is subsequently fermented to ethanol by means of anaerobic microbial catalysts known as homoacetogens. As a homoacetogen, Clostridium autoethanogenum is able to metabolize syngas/CO and synthesize ethanol, but limited work has been accomplished with it and ethanol concentration achieved was low. In order to improve its ethanol production, fermentation process and some factors affecting cell growth or product formation were studied. Although xylose is an easily-used carbon source for C. autoethanogenum, results showed that the primary end product in xylose metabolism was acetate, while ethanol remained in a low level even with high level of xylose (8-10 g/L). C. autoethanogenum grew rapidly in the medium containing xylose and organic nitrogen source, and high cell density was achieved. When nitrogen source was switched to NH4Cl, C. autoethanogenum grew much slowly and the overall cell density diminished. However, nitrogen source didn’t have much influence on ethanol production if xylose was used as the carbon source. Two-step fermentation, i.e., growing on xylose (first stage) and then fermenting with CO (second stage), was performed in a 3 L bioreactor to study the batch fermentation of C. autoethanogenum. Results indicated that cell growth and acetate production occurred in the first stage, whereas ethanol was primarily produced in the second stage when xylose was exhausted and CO became the sole carbon source. Fermentation curves showed that pH value and oxidation-reduction potential (ORP) dropped with cell growth and acetate production, while ethanol production was accompanied by the decrease of acetate concentration and the rise of pH value and ORP. CO2 evolution was observed in both growth and fermentation stages, and a small amount of H2 was detected in the outlet gas during the CO fermentation stage. But due to the limitations of bioreactor and operating conditions, gas-liquid mass transfer in the bioreactor was poor, which resulted in the low efficiency of CO utilization, and only 1.71 g/L ethanol was obtained. To eliminate the limitations, the bioreactor was modified and equipped with a specific device which could keep a constant pressure in the headspace. One-step fermentation was carried out in the modified bioreactor using CO as the sole carbon and energy source. In spite of reduced growth rate and cell density, C. autoethanogenum produced more ethanol than it did in two-step fermentation. The maximum ethanol concentration obtained was 7.36 g/L, much greater than that reported in the previous studies. Moreover, during the whole fermentation process, acetate concentration remained lower than 1.1 g/L. Summarily, the results suggest that C. autoethanogenum is a promising strain in ethanol production from CO, and this study presents a reference for the scale-up of CO fermentation to ethanol with C. autoethanogenum. [ABSTRACT FROM AUTHOR]

In bioindustry, syngas fermentation is a promising technology for biofuel production without the use of plant biomass as sugar-based feedstock. The aim of this study was to identify optimal conditions for high efficient ethanol and volatile fatty acids (VFA) production from synthetic gas fermentation. Therefore, the effect of different gases (pure CO, H 2 , and a synthetic syngas mixture), media (acetate medium and acetate-free medium), and biocatalyst (pure and mixed culture) were studied. Acetate was the most dominant product independent on inoculum type. The maximum concentration of volatile fatty acids and ethanol was achieved by the pure culture ( Clostridium ragsdalei ). Depending on the headspace gas composition, VFA concentrations were up to 300% higher after fermentation with Clostridium ragsdalei compared to fermentation with mixed culture. The preferred gas composition with respect to highest VFA concentration was pure CO (100%) regardless of microbial composition of the inoculum and media composition. The addition of acetate had a negative impact on the VFA formation which was depending on the initial gas composition in head space. [ABSTRACT FROM AUTHOR]

This study presents the design and assessment of site-specific supply chains and related manufacturing processes for the production of bio-based chemicals from the syngas platform and via gasification of lignocellulosic biomass followed by syngas fermentation. The supply chains include feedstocks production and collection, biomass gasification, syngas fermentation, and downstream processing. For each of these stages, different alternatives were considered: four feedstocks (pine, corn stover, sugarcane bagasse, and eucalyptus), three products (ethanol, 2,3-butanediol and hexanoic acid), and three geographical locations (the Netherlands, the USA, and Brazil). Conceptual development and analysis of the supply chains were done through the combination of different design and assessment tools, namely biomass supply chains design, fermentation process design (based on thermodynamics and transport), process simulation, and economic and environmental assessments. The minimum selling price (MSP) and two environmental impact categories, i.e., global warming potential (GWP) and non-renewable energy use (NREU), were used as performance indicators. These were compared to data reported in scientific literature and commercial sources for similar processes and ­products. The best overall performance was obtained for the production of 2,3-butanediol from pine sourced in the USA. In the cases of ethanol and hexanoic acid, the syngas fermentation stage had significant contributions to MSP, GWP, and NREU, due mainly to its high energy requirements. Regarding the geographical location, the best economic performance was obtained for the USA followed by the Netherlands and Brazil respectively. Furthermore, operation in Brazil led to the lowest environmental impacts, followed by the Netherlands and the USA. © 2017 Society of Chemical Industry and John Wiley & Sons, Ltd [ABSTRACT FROM AUTHOR]

Bioconversion of CO/syngas to produce ethanol is a novel route in bioethanol production, which can be accomplished by some acetogens. Specific culture vessels and techniques are needed to cultivate these microorganisms since they are anaerobic and substrates are gaseous. In this work, gas-sampling bag was applied as a gas-cultivation system to study CO/syngas bioconversion by Clostridium autoethanogenum and was demonstrated to be efficient because of its flexibility and excellent ability to maintain the headspace atmosphere. C. autoethanogenum can use CO as the sole carbon and energy source to produce ethanol, acetate as well as CO 2 . In the experimental range, higher ethanol production was favored by higher yeast extract concentrations, and the maximum ethanol concentration of 3.45 g/L was obtained at 1.0 g/L of yeast extract. Study with various bottled gases showed that C. autoethanogenum preferred to use CO other than CO 2 and produced the highest level of ethanol with 100% CO as the substrate. C. autoethanogenum can also utilize biomass-generated syngas (36.2% CO, 23.0% H 2 , 15.4% CO 2 , 11.3% N 2 ), but the process proceeded slowly and insufficiently due to the presence of O 2 and C 2 H 2 . In our study, C . autoethanogenum showed a better performance in the bioconversion of CO to ethanol than Clostridium ljungdahlii , a strain which has been most studied, and for both strains, ethanol production was promoted by supplementing 0.5 g/L of acetate. [ABSTRACT FROM AUTHOR]

Background: The product of current syngas fermentation systems is an ethanol/acetic acid mixture and the goal is to maximize ethanol recovery. However, ethanol currently has a relatively low market value and its separation from the fermentation broth is energy intensive. We can circumvent these disadvantages of ethanol production by converting the dilute ethanol/acetic acid mixture into products with longer carbon backbones, which are of higher value and are more easily extracted than ethanol. Chain elongation, which is the bioprocess in which ethanol is used to elongate short-chain carboxylic acids to medium-chain carboxylic acids (MCCAs), has been studied with pure cultures and open cultures of microbial consortia (microbiomes) with several different substrates. While upgrading syngas fermentation effluent has been studied with open cultures, to our knowledge, no study exists that has performed this with pure cultures. Results: Here, pure cultures of Clostridium kluyveri were used in continuous bioreactors to convert ethanol/acetic acid mixtures into MCCAs. Besides changing the operating conditions in regards to substrate loading rates and composition, the effect of in-line product extraction, pH, and the use of real syngas fermentation effluent on production rates were tested. Increasing the organic loading rates resulted in proportionally higher production rates of n-caproic acid, which were up to 40 mM day-1 (4.64 g L-1 day-1) at carbon conversion efficiencies of 90% or higher. The production rates were similar for bioreactors with and without in-line product extraction. Furthermore, a lower ethanol/acetic acid ratio (3:1 instead of 10:1) enabled faster and more efficient n-caproic acid production. In addition, n-caprylic acid production was observed for the first time with C. kluyveri (up to 2.19 ± 0.34 mM in batch). Finally, the use of real effluent from syngas fermentation, without added yeast extract, but with added defined growth factors, did maintain similar production rates. Throughout the operating period, we observed that the metabolism of C. kluyveri was inhibited at a mildly acidic pH value of 5.5 compared to a pH value of 7.0, while reactor microbiomes perform successfully at mildly acidic conditions. Conclusions: Clostridium kluyveri can be used as a biocatalyst to upgrade syngas fermentation effluent into MCCAs at pH values above 5.5. [ABSTRACT FROM AUTHOR]

Biomass co-gasification and syngas fermentation is one of the least trotted paths in the Indian biofuel production scenario. The aim of the research here is to develop an equilibrium model of biomass co-gasification and syngas fermentation in Aspen plus. Two Indian lignocellulosic biomass namely wheat straw and sugarcane bagasse have been selected as feedstock for this co-gasification process followed by the syngas fermentation. The process shows promising results in terms of ethanol production and energy usage. The effects of gasification process parameters on overall process performance have been analysed. Empirical regression equations were constructed to account for the effects of biomass ratio, which is the ratio of the masses of wheat straw and sugarcane bagasse, gasification temperature and equivalence ratio over ethanol yield, lower heating value of syngas, cold gas efficiency, overall system efficiency, and CO 2 emission using response surface methodology. Finally, a multi-objective optimization has been conducted to maximize the ethanol production and overall system efficiency, and to minimize the CO 2 emission of the system. Results suggest that the wheat straw and sugarcane bagasse ratio of 4:1, 919.5 o C temperature and 0.15 equivalence ratio is the optimum point with an ethanol production rate of 20.917 tonnes/hr, overall efficiency of 52.184%, and a minimum CO 2 emission of 32669.862 kg/h. • Indian lignocellulosic biomass was used as feedstock for the hybrid process. • Equilibrium process modelling was done using Aspen plus software. • Regression equations were proposed for various performance parameters in terms of gasification input parameters. • Multi-objective optimization was conducted to maximize ethanol yield and energy efficiency. • Multi-objective optimization was conducted to minimize CO2 emission. [ABSTRACT FROM AUTHOR]

Gasification of lignocellulosic biomass followed by syngas fermentation is a promising process for producing fuels and chemicals. Syngas fermentation, however, is commonly limited by low mass transfer rates. In this work, a horizontally oriented rotating packed bed (h-RPB) reactor was developed to improve mass transfer and enhance ethanol production. In the h-RPB reactor, cell attachment materials were packed in the reactor and half submerged in the liquid and half exposed to the headspace. With continuous rotation of the packing materials, the cells in biofilm were alternately in contact with liquid and headspace; thus, transport of syngas to the cells occurred in both the liquid phase and headspace. The volumetric mass transfer coefficient (k L a) of the h-RPB reactor was lower than that in a traditional continuous stirred tank reactor (CSTR), indicating the mass transfer in the liquid phase of h-PRB was lower than CSTR, and the mass transfer in the headspace phase played an important role in syngas fermentation. The syngas fermentation of Clostridium carboxidivorans P7 in h-RPB resulted in a 7.0 g/L titer and 6.7 g/L/day productivity of ethanol, respectively, 3.3 times higher than those obtained in a CSTR under the same operational conditions. The results demonstrate that the h-RPB reactor is an efficient system for syngas fermentation, making cellulosic ethanol biorefinery one step closer to technical and economic feasibility. [ABSTRACT FROM AUTHOR]

Clostridium carboxidivorans P7 is a strict anaerobic bacterium capable of converting syngas to biofuels. However, its fermentation profiles is poorly understood. Here, various end-products, including acetic acid, butyric acid, hexanoic acid, ethanol and butanol were supplemented to evaluate their effects on fermentation profiles in C . carboxidivorans at two temperatures. At 37 °C, fatty acids addition likely led to more corresponding alcohols production. At 25 °C, C2 and C4 fatty acids supplementation resulted in more corresponding higher fatty acids, while supplemented hexanoic acid increased yields of C2 and C4 fatty acids and hexanol. Supplementation of ethanol or butanol caused increased production of C2 and C4 acids at both temperatures; however, long-chain alcohols were still more likely produced at lower temperature. In conclusion, fermentation profiles of C . carboxidivorans can be changed in respond to pre-added end-products and carbon flow may be redirected to desired products by controlling culture conditions. [ABSTRACT FROM AUTHOR]

Lowering the redox potential of the growth medium is a critical step in the cultivation of obligate anaerobes for production of biofuels from synthesis gas. In this study, the simultaneous effects of reducing solutions (sodium sulfide and/or cysteine-HCl) and initial medium pH on the fermentation of synthesis gas usingClostridium ljungdahliiwas investigated. The results suggested the plausible provision of more electrons into the culture in the presence of 5.07 mM cysteine-HCl at the medium pH 5.9. This culture demonstrated enhanced ethanol production (48%) and ethanol to acetate production ratio (24%) compared to the cells cultivated in standard growth medium recommended by ATCC. [ABSTRACT FROM PUBLISHER]

[Display omitted] • Detailed analysis on gas transfer rate in industrial syngas fermentation. • CFD reveals mass transfer limitation topography in external-loop gas-lift reactors. • Bubbles smaller than 2 mm are key to reach required mass transfer capacities. • The produced ethanol minimizes bubble size and increases gas hold-up. • Transport limitations in air–water systems can be alleviated in syngas fermentation. Mass transfer limitations in syngas fermentation processes are mostly attributed to poor solubility of CO and H 2 in water. Despite these assumed limitations, a syngas fermentation process has recently been commercialized. Using large-sale external-loop gas-lift reactors (EL-GLR), CO-rich off-gases are converted into ethanol, with high mass transfer performance (7–8.5 g.L-1.h−1). However, when applying established mass transfer correlations, a much poorer performance is predicted (0.3–2.7 g.L-1.h−1). We developed a CFD model, validated on pilot-scale data, to provide detailed insights on hydrodynamics and mass transfer in a large-scale EL-GLR. As produced ethanol could increase gas hold-up (+30%) and decrease the bubble diameter (≤2 mm) compared to air–water mixtures, we found with our model that a high volumetric mass transfer coefficient (650–750 h−1) and mass transfer capacity (7.5–8 g.L-1.h−1) for CO are feasible. Thus, the typical mass transfer limitations encountered in air–water systems can be alleviated in the syngas-to-ethanol fermentation process. [ABSTRACT FROM AUTHOR]

• Biomass-to-ethanol via syngas fermentation with solid-oxide fuel cell system is designed. • Optimized gasification and fermentation condition obtained for ethanol production. • Optimal configuration of system scale is determined by multi-objective optimization. • Production cost reach 430.1 €/t ethanol with annual 69,595 tons from 200,000 tons woodchips. • Biomass-to-ethanol with solid oxide fuel cell system achieve efficiency of 55% Bioethanol is regarded as a high-quality and clean liquid fuel with a renewable production process through bioconversion and chemical synthesis technology. Syngas fermentation route has received increasing attention due to mild reaction conditions and the elimination of expensive pretreatment and enzyme. To improve the economy and energy efficiency of the bioethanol production, a novel combined biomass gasification-syngas fermentation-solid oxide fuel cell system is proposed in this work. The techno-economic assessment is performed by the parametric analysis of the gasification and fermentation process. The sensitivity analysis shows that the dual fluidized bed with woodchips has the highest CO and H 2 content, and the ethanol yield can be enhanced by improving the substrate conversion rate. A tentative economic assessment is performed with a total production cost of 430.1 €/t ethanol, revealing the economy of the hybrid gasification-syngas fermentation process. Besides, the problem of syngas distribution and fermenter exhaust recycling is further explored by introducing the SOFC system to realize the cascade utilization of energy. The trade-off between the combined system efficiency, total annual cost, and product price is conducted by multi-objective optimization with the technique for order of preference by similarity to ideal solution (TOPSIS) method. The optimized system can reach the system efficiency of 55%, the minimum total annual cost of 19.9 M€, and the total product price of 64.5 M€. The proposed system could demonstrate the feasibility of bioethanol production via biomass syngas fermentation with a high-efficiency and cost-competitive pathway. [ABSTRACT FROM AUTHOR]

Syngas fermentation is a promising process for producing fuels and chemicals from lignocellulosic biomass. Currently syngas fermentation faces several engineering challenges, with gas-to-liquid mass transfer limitation representing the major bottleneck. The aim of this work is to evaluate the performance of a monolithic biofilm reactor (MBR) as a novel reactor configuration for syngas fermentation. The volumetric mass transfer coefficient ( k L a ) of the MBR was evaluated in abiotic conditions within a wide range of gas flow rates (i.e., gas velocity in monolithic channels) and liquid flow rates (i.e., liquid velocity in the channels). The k L a values of the MBR were higher than those of a controlled bubble column reactor (BCR) in certain conditions, due to the slug flow pattern in the monolithic channels. A continuous syngas fermentation using Clostridium carboxidivorans P7 was conducted in the MBR system under varying operational conditions, with the variables including syngas flow rate, liquid recirculation between the monolithic column and reservoir, and dilution rate. It was found that the syngas fermentation performance – measured by such parameters as syngas utilization efficiency, ethanol concentration and productivity, and ratio of ethanol to acetic acid – depended not only on the mass transfer efficiency but also on the biofouling or abrading of the biofilm attached on the monolithic channel wall. At a condition of 300 mL/min of syngas flow rate, 500 mL/min of liquid flow rate, and 0.48 day − 1 of dilution rate, the MBR produced much higher syngas (CO/H 2 ) utilization efficiency and much greater metabolite (ethanol/acetic acid) productivity than what was obtained using a traditional bubble column reactor. The study demonstrates the great potential of MBR as a promising reactor configuration for syngas fermentation with high mass transfer efficiency, low energy consumption, and high metabolite productivity. [ABSTRACT FROM AUTHOR]

Highlights: [•] Low mass transfer efficiency is one major limitation in syngas fermentation. [•] Hollow fiber membrane (HFM) reactor leads to a high mass transfer efficiency. [•] Syngas fermentation of Clostridium carboxidivoran P7 in HFM reactor was successfully established. [•] Syngas fermentation in HFM generated a higher ethanol titer than suspended culture. [•] Syngas flow rate and liquid recirculation rate are crucial for HFM operation. [ABSTRACT FROM AUTHOR]