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The presence of lignocellulose-derived microbial inhibitory compounds (LDMICs) in lignocellulosic biomass (LB) hydrolysates is a barrier to efficient conversion of LB hydrolysates to fuels and chemicals by fermenting microorganisms. Results from this study provide convincing evidence regarding the effectiveness of metabolically engineered

Microorganisms used in syngas fermentation require nutrients to grow and convert syngas (CO, H2 and CO2) into various products. Many of the essential nutrients can be provided by biochar. Poultry litter biochar (PLBC) contains minerals and trace metals and has a high pH buffering capacity, making it suitable as a nutrient supplement. The effects of PLBC loadings from 1 to 20 g L−1 on syngas fermentation were determined in 250 ml bottle assays. Results showed that 10 and 20 g L−1 PLBC significantly increased ethanol production compared to standard yeast extract (YE) medium. Fermentations in a 3L continuous stirred tank reactor (CSTR) with 10 g L−1 PLBC with and without 4-morpholineethanesulfonic acid (MES) showed 64% and 36% more ethanol production, respectively, than standard medium. The acetic acid accumulated at the beginning of fermentation was completely converted to ethanol in all media tested in the CSTR. These results demonstrate the feasibility of using PLBC medium without costly MES in the CSTR to enhance ethanol production from syngas for potential use at commercial scale.

Lignocellulosic biomass such as switchgrass can be converted to n-butanol using fermentation, which can be further processed into jet fuel. Traditional acetone-butanol-ethanol (ABE) fermentation only converts sugars derived from switchgrass to ABE. Novel co-fermentation processes convert sugars and gas (CO

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:H2:CO2 (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 H2 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.

An A. nidulans strain with a pyridoxine marker was used for continuous production of aryl alcohol oxidase (AAO) in a trickle bed reactor (TBR). Modified medium with reduced zinc, no copper, and 5 g/L ascorbic acid that reduced melanin production and increased AAO productivity under growth limited conditions was used. Two air flow rates, 0.11 L/min (0.1 vvm) and 1.1 L/min (1.0 vvm) were tested. More melanin formation and reduced protein productivity were observed with air flow rate of 1.1 L/min. Three random packings were used as support for the fungus inside the TBR column, two of which were hydrophobic and one which was hydrophilic, and three different dilution rates were tested. The use of GEA BCN 030 hydrophobic packing resulted in greater AAO yield and productivity than the other packings. Increasing dilution rates favored melanin formation and citric, lactic and succinic acid accumulation, which decreased AAO yield and productivity.

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 250 mL bottle reactors at 150 rpm and 37 °C with syngas containing CO:H2:CO2 (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 H2 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.

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 1200tons 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 36.5million 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.

Lignocellulosic biomass such as switchgrass can be converted to n-butanol using fermentation, which can be further processed into jet fuel. Traditional acetone-butanol-ethanol (ABE) fermentation only converts sugars derived from switchgrass to ABE. Novel co-fermentation processes convert sugars and gas (CO2/H2) produced during fermentation into butanol, thus increasing ABE yields by 15.5% compared to traditional ABE fermentation. Herein, the environmental impact of a Switchgrass to Jet Fuel (STJ) pathway was assessed using life cycle assessment (LCA) from well-to-wake. LCAs were performed for greenhouse gas (GHG) emissions from jet fuel production via co-fermentation of sugars and gas for ideal and practical cases of ABE fermentation and seven other jet fuel pathways. The ideal case assumes 100% sugar recovery and 95% ABE yield. The practical case assumes 90% sugar recovery and an 80% ABE yield. Results are presented based on 100-year global warming potential (GWP) per MJ of jet fuel. Co-products were allocated using various methods. The increase in butanol yield via the co-fermentation technology reduced GWP-100 for the STJ pathway by 6.5% compared to traditional ABE fermentation. Similarly, the STJ pathway for the practical case with co-fermentation had 14.2%, 47.5%, 73.8%, and 44.4% less GWP-100 compared to HRJ, Fischer-Tropsch jet fuel from switchgrass, Fischer-Tropsch jet fuel from coal, and conventional petroleum jet fuel. The results demonstrate that the STJ pathway via co-fermentation has the potential to increase product yield while reducing GHG emissions compared to other jet fuel production pathways.

Biochar is traditionally used to improve soil properties in arable land and as adsorbent or precursor of activated carbon in wastewater treatment. Recent advances have shown biochar potentials in enhancing productions of biofuels and chemicals such as bio-ethanol, butanol, methane, hydrogen, bio-diesel, hydrocarbons and carboxylic acids. The properties of biochar such as high levels of porosity, functional groups, cation exchange capacity, pH buffering capacity, electron conductivity, and macro-/micro- nutrients (Na, K, Ca, Mg, P, S, Fe, etc.) provide appropriate conditions to relieve physicochemical stresses on microorganisms through pH buffering, detoxification, nutrients supply, serving as electron carrier and supportive microbial habitats. This paper critically reviewed biochar production and characteristics, biochar utilization in anaerobic digestion, composting, microbial fermentation, hydrolysate detoxification, catalysis in biomass refinery and biodiesel synthesis. This review provides novel vision of biochar application, which could guide future research towards cleaner and more economic production of renewable fuels and bio-based chemicals.

Syngas (mixture of CO, H2 and CO2) fermentation suffers from mass transfer limitation due to low solubility of CO and H2 in the liquid medium. Therefore, it is critical to characterize the mass transfer in syngas fermentation reactors to guide in delivery of syngas to the microorganisms. The objective of this study is to measure and predict the overall volumetric mass transfer coefficient, kLa for O2 at various operating conditions in a 7-L sparged and non-sparged continuous stirred-tank reactor (CSTR). Measurements indicated that the kLa for O2 increased with an increase in air flow rate and agitation speed. However, kLa for O2 decreased with the increase in the headspace pressure. The highest kLa for O2 with air sparged in the CSTR was 116 h−1 at 600 sccm, 900 rpm, 101 kPa, and 3 L working volume. Backmixing of the headspace N2 in the sparged CSTR reduced the observed kLa. The mass transfer model predicted the kLa for O2 within 10% of the experimental values. The model was extended to predict the kLa for syngas components CO, CO2 and H2, which will guide in selecting operating conditions that minimize power input to the bioreactor and maximize the syngas conversion efficiency.

Synthesis gas (syngas) can be produced via gasification of diverse feedstocks. Syngas is used as a “fermentation” feedstock (carbon and energy sources) for the biological production of liquid fuels and plethora of platform chemicals. The approach has been extensively studied in a lab scale, and efforts have been made to commercialize syngas fermentation technology. There are, however, several challenges such as low yield of syngas fermenting microorganisms, mass transfer limitation of syngas, substrate and product inhibition, and cost of products separation and recovery among others. This chapter presents a critical review on syngas quality and impurities, selected syngas utilizing microorganisms, microbial reactions and pathways, C1 chemistry, fermentation products, and approaches to overcome mass transfer, and kinetic limitations. The current status of the syngas fermentation technology and future research perspectives in syngas fermentation to biofuels and biochemicals is also included.

Syngas fermentation is a promising technique to produce biofuels using syngas obtained through gasified biomass and other carbonaceous materials or collected from industrial CO-rich off-gases. The primary components of syngas, carbon monoxide (CO) and hydrogen (H2), are converted to alcohols and other chemicals through an anaerobic fermentation process by acetogenic bacteria. Dissolved CO and H2 concentrations in fermentation media are among the most important parameters for successful and stable operation. However, the difficulties in timely and precise dissolved CO and H2 measurements hinder the industrial-scale commercialization of this technique. The purpose of this article is to provide a comprehensive review of available dissolved CO and H2 measurement methods, focusing on their detection mechanisms, CO and H2 cross interference and operations in syngas fermentation process. This paper further discusses potential novel methods by providing a critical review of gas phase CO and H2 detection methods with regard to their capability to be modified for measuring dissolved CO and H2 in syngas fermentation conditions.

An efficient syngas fermentation bioreactor provides a mass transfer capability that matches the intrinsic kinetics of the microorganism to obtain high gas conversion efficiency and productivity. In this study, mass transfer and gas utilization efficiencies of a trickle bed reactor during syngas fermentation by Clostridium ragsdalei were evaluated at various gas and liquid flow rates. Fermentations were performed using a syngas mixture of 38% CO, 28.5% CO2, 28.5% H2 and 5% N2, by volume. Results showed that increasing the gas flow rate from 2.3 to 4.6sccm increased the CO uptake rate by 76% and decreased the H2 uptake rate by 51% up to Run R6. Biofilm formation after R6 increased cells activity with over threefold increase in H2 uptake rate. At 1662h, the final ethanol and acetic acid concentrations were 5.7 and 12.3g/L, respectively, at 200ml/min of liquid flow rate and 4.6sccm gas flow rate.

Gasification-fermentation is a thermochemical-biological platform for the production of fuels and chemicals. Biomass is gasified at high temperatures to make syngas, a gas composed of CO, CO2, H2, N2 and other minor components. Syngas is then fed to anaerobic microorganisms that convert CO, CO2 and H2 to alcohols by fermentation. This platform offers numerous advantages such as flexibility of feedstock and syngas composition and lower operating temperature and pressure compared to other catalytic syngas conversion processes. In comparison to hydrolysis-fermentation, gasification-fermentation has a major advantage of utilizing all organic components of biomass, including lignin, to yield higher fuel production. Furthermore, syngas fermentation microorganisms do not require strict CO:H2:CO2 ratios, hence gas reforming is not required. However, several issues must be addressed for successful deployment of gasification-fermentation, particularly those that involve the integration of gasification and fermentation. Most previous reviews have focused only on either biomass gasification or syngas fermentation. In this review, the critical factors that affect the integration of biomass gasification with syngas fermentation, such as carbon conversion efficiency, effect of trace gaseous species, H2 to CO ratio requirements, and microbial preference of carbon substrate, are thoroughly discussed.

Biochar can be an inexpensive pH buffer and source of mineral and trace metal nutrients in acetone-butanol-ethanol (ABE) fermentation. This study evaluated the feasibility of replacing expensive 4-morpholineethanesulfonic acid (MES) P2 buffer and mineral nutrients with biochar made from switchgrass (SGBC), forage sorghum (FSBC), redcedar (RCBC) and poultry litter (PLBC) for ABE fermentation. Fermentations using Clostridium beijerinckii ATCC 51743 in glucose and non-detoxified switchgrass hydrolysate media were performed at 35 °C in 250 mL bottles for 72 h. Medium containing buffer and minerals without biochar was the control. Similar ABE production (about 18.0 g/L) in glucose media with SGBC, FSBC and RCBC and control was measured. However in non-detoxified switchgrass hydrolysate medium, SGBC, RCBC and PLBC produced more ABE (about 18.5 g/L) than the control (10.1 g/L). This demonstrates that biochar is an effective buffer and mineral supplement for ABE production from lignocellulosic biomass without costly detoxification process.

Syngas fermentation for producing biofuels and other products suffers from mass transfer limitations due to low CO and H2 solubility in liquid medium. Therefore, it is critical to characterize mass transfer rates of these gases to guide bioreactor design and optimization. This work presents a novel technique to measure the volumetric mass transfer coefficients (kia) for H2 and CO using gas chromatography in a non-porous hollow fiber reactor (HFR). The largest measured kia for H2 and CO were 840 and 420 h−1, respectively. A model was developed to predict kia for H2 and CO that agreed well with experimental data. This study is the first to measure, compare, and model both H2 and CO mass transfer coefficients in an HFR. Based on model predictions, HFRs have the potential to be a reactor of choice for syngas fermentation as a result of high mass transfer that can support high cell densities.

The present study evaluated butanol production from switchgrass based on hydrothermolysis pretreatment. The inhibitors present in the hydrolyzates were measured. Results showed poor butanol production (1g/L) with non-detoxified hydrolyzate. However, adjusting the pH of the non-detoxified hydrolyzate to 6 and adding 4 g/L CaCO3 increased butanol formation to about 6g/L. There was about 1g/L soluble lignin content (SLC), and various levels of furanic and phenolic compounds found in the non-detoxified hydrolyzate. Detoxification of hydrolyzates with activated carbon increased the butanol titer to 11 g/L with a total acetone, butanol and ethanol (ABE) concentration of 17 g/L. These results show the potential of butanol production from hydrothermolysis pretreated switchgrass.

A trickle bed reactor (TBR) with recycle was designed and tested using Aspergillus nidulans with a pyridoxine marker and over-expressing/secreting recombinant client xylanase B (XynB). The pyridoxine marker prevented the fungus from synthesizing its own pyridoxine and fungus was unable to grow when no pyridoxine was present in the medium; however, enzyme production was unaffected. Uncontrolled mycelia growth that led to clogging of the TBR was observed when fungus without a pyridoxine marker was used for XynB production. Using the fungus with pyridoxine marker, the TBR was operated continuously for 18 days and achieved a XynB output of 41 U/ml with an influent and effluent flow rate of 0.5 ml/min and a recycle flow rate of 56 ml/min. Production yields in the TBR were 1.4 times greater than a static tray culture and between 1.1 and 67 times greater than yields for SSF enzyme production stated in the literature.

Biomass and waste materials are converted to ethanol by gasification followed by fermentation of the synthesis gas using acetogenic bacteria such as Clostridium ragsdalei. Fuel ethanol is a high-volume, low-cost commodity, so capital and operating expenses must be very low to achieve profitable operation. The fermentation medium supplies minerals, metals, and organic co-factors needed to build cell biomass and the functional enzyme platform that conducts the conversion reactions. Research directed toward commercial deployment of synthesis gas (syngas) fermentation processes must consider cost and availability of medium components, and use economically viable medium. A method based on elemental accounting of cell composition was developed to assess the cost and function of fermentation media and their potential for commercial use for syngas fermentation to produce ethanol using C. ragsdalei. Cost of medium was reduced from $9.87 L -1 to $0.26 L -1 . Additional cost reduction can be achieved by eliminating components that are not required, using commercial components purchased at bulk commodity prices, using H 2 S and NH 3 from the syngas production in lieu of purchased sodium sulfide and ammonium chloride, and retaining the acetate buffer from ethanol distillation. These savings can reduce the medium cost below $0.01 L -1 .

Eastern redcedar is an invasive softwood species in Oklahoma and across grasslands in the Central Plains of the United States and potential feedstock for butanol production. Butanol has higher energy content than ethanol and can be upgraded to jet and diesel fuels. The objective of this study was to develop a process for production of butanol from redcedar. Results showed that Clostridium acetobutylicum ATCC 824 and Clostridium beijerinckii NCIMB 8052 did not grow in fermentation medium with citrate buffer. However, both strains grew in the medium with acetate buffer, resulting in 3–4 g/L greater butanol than without acetate. Detoxification of redcedar hydrolyzate was required to increase butanol concentration from 1 to 13 g/L. Hydrolyzate was detoxified by activated carbon to remove inhibitors. Fermentations in detoxified redcedar hydrolyzate reached 13 g/L butanol and 19 g/L total ABE, comparable to glucose control. This shows the potential for redcedar use in butanol production.

A trickle-bed reactor (TBR) when operated in a trickle flow regime reduces liquid resistance to mass transfer because a very thin liquid film is in contact with the gas phase and results in improved gas–liquid mass transfer compared to continuous stirred tank reactors (CSTRs). In the present study, continuous syngas fermentation was performed in a 1-L TBR for ethanol production by Clostridium ragsdalei. The effects of dilution and gas flow rates on product formation, productivity, gas uptakes and conversion efficiencies were examined. Results showed that CO and H2 conversion efficiencies reached over 90% when the gas flow rate was maintained between 1.5 and 2.8 standard cubic centimeters per minute (sccm) at a dilution rate of 0.009 h−1. A 4:1 molar ratio of ethanol to acetic acid was achieved in co-current continuous mode with both gas and liquid entered the TBR at the top and exited from the bottom at dilution rates of 0.009 and 0.012 h−1, and gas flow rates from 10.1 to 12.2 sccm and 15.9 to 18.9 sccm, respectively.

An Aspergillus nidulans cell factory was genetically engineered to produce an aryl alcohol oxidase (AAO). The cell factory initiated production of melanin when growth-limited conditions were established using stationary plates and shaken flasks. This phenomenon was more pronounced when the strain was cultured in a trickle bed reactor (TBR). This study investigated different approaches to reduce melanin formation in fungal mycelia and liquid medium in order to increase the enzyme production yield. Removal of copper from the medium recipe reduced melanin formation in agar cultures and increased enzyme activities by 48% in agitated liquid cultures. Copper has been reported as a key element for tyrosinase, an enzyme responsible for melanin production. Ascorbic acid (0.44 g/L) stopped melanin accumulation, did not affect growth parameters and resulted in AAO activity that was more than two-fold greater than a control treatment with no ascorbic acid.

Lignocellulosic biomass is a vast and underutilized resource for the production of sugars and biofuels. However, the structural complexity of lignocellulosic biomass and the need for multiple pretreatment and enzymatic steps for sugar release renders this process economically challenging. Here, we report a novel approach for direct, single container, exogenous enzyme-free conversion of lignocellulosic biomass to sugars and biofuels using the anaerobic fungal isolate strain C1A. This approach utilizes simple physiological manipulations for timely inhibition and uncoupling of saccharolytic and fermentative capabilities of strain C1A, leading to the accumulation of sugar monomers (glucose and xylose) in the culture medium. The produced sugars, in addition to fungal hyphal lysate, are subsequently converted by Escherichia coli strain K011 to ethanol. Using this approach, we successfully recovered 17.0% (w/w) of alkali-pretreated corn stover (20.0% of its glucan and xylan content) as sugar monomers in the culture media. More importantly, 14.1% of pretreated corn stover (17.1% of glucan and xylan content) was recovered as ethanol at a final concentration of 28.16 mM after the addition of the ethanologenic strain K011. The high ethanol yield obtained is due to its accumulation as a minor fermentation end product by strain C1A during its initial growth phase, the complete conversion of sugars to ethanol by strain K011, and the possible conversion of unspecified substrates in the hyphal lysate of strain C1A to ethanol by strain K011. This study presents a novel, versatile, and exogenous enzyme-free strategy that utilizes a relatively unexplored group of organisms (anaerobic fungi) for direct biofuel production from lignocellulosic biomass.

Ethanol and fructose are important materials for the fuel and food markets; high fructose syrup production processes are still costly. The effect of inoculum medium composition on the production of fructose and ethanol from dates by selective fermentation was investigated. The addition of minerals improved the fructose content in the produced syrup and ethanol productivity. The addition of malt extract to the medium increased fructose yield from 94% to 97%, but decreased ethanol yield from 78% to 69%. The presence of peptone increased ethanol productivity and fructose fraction in sugar, while iron increased fructose yield to 100% in syrups with 371 g initial sugar/L. Compared to the commonly marketed 55% fructose, selective fermentation of date extract produced sugar syrups with at least 82% fructose and significant amounts of ethanol. The excellent fit by the new expanded model prediction to the experimental data makes them valuable tools for further process development or industrial applications.

Essential oils from plants are known to be inhibitory to yeast and amylase activity. This study investigated the effects of oil in Eastern redcedar ( Juniperus virginiana L .) on enzymatic cellulose hydrolysis and yeast fermentation for ethanol production from redcedar. Enzymatic hydrolysis was conducted with 2% and 4% (w/w) cellulose loadings, 25 FPU/g cellulose and 50 FPU/g cellulose enzyme loadings (Accelerase ® 1500) and 0%, 0.5% and 1% (w/w) redcedar oil loadings. The presence of 0.5% and 1% (w/w) redcedar oil inhibited cellulose-to-glucose yields by 26% and 33% and inhibited cell growth and ethanol production by Saccharomyces cerevisiae during the first 9 h. Similar ethanol concentrations of up to 22 g/l (92% of theoretical yield) were produced with all treatments within 12 h. The results of this study suggest that redcedar oil be removed prior to enzymatic hydrolysis and fermentation for efficient ethanol production using redcedars.

Syngas fermentation to fuels is a technology on the verge of commercialization. Low cost of fermentation medium is important for process feasibility. The use of corn steep liquor (CSL) instead of yeast extract (YE) in Alkalibaculum bacchi strain CP15 bottle fermentations reduced the medium cost by 27% and produced 78% more ethanol. When continuous fermentation was performed in a 7-L fermentor, 6 g/L ethanol was obtained in the YE and YE-free media. When CSL medium was used in continuous fermentation, the maximum produced concentrations of ethanol, n-propanol and n-butanol were 8 g/L, 6 g/L and 1 g/L, respectively. n-Propanol and n-butanol were not typical products of strain CP15. A 16S rRNA gene-based survey revealed a mixed culture in the fermentor dominated by A. bacchi strain CP15 (56%) and Clostridium propionicum (34%). The mixed culture presents an opportunity for higher alcohols production from syngas.

Syngas is produced by thermochemical conversion, e.g., pyrolysis and gasification, of biomass, animal waste, coal, municipal solid waste and other carbonaceous materials, or directly from CO-rich off-gases from industry, e.g. steel mills. Syngas components (mainly CO, H2 and CO2) are converted to alcohols and other chemicals by acetogenic bacteria through the Wood-Ljungdahl pathway or its derivatives. Syngas fermentation is affected by the acetogen(s), type of reactor, gas composition, medium components, operating parameters, gas-liquid mass transfer and fermentation strategies. These factors affect product distribution, titer, yield, productivity and process feasibility. In this article, syngas fermentation process development with focus on microorganisms, gas composition, medium design, gas-liquid mass transfer fermentation strategies, techno-economic analysis and commercialization efforts are critically reviewed. This review provides new insights in syngas fermentation, which can guide future research towards commercialization of renewable and sustainable biofuels and chemicals.

In this study, the economic feasibility of producing ethanol from gasification followed by syngas fermentation via commercially available technologies was theoretically evaluated using a set of selected livestock and agricultural and forest residuals ranging from low valued feedstocks (i.e., wood, wheat straw, wheat straws blended with dewatered swine manure, and corn stover) to high valued oilseed rape meal. A preliminary cost analysis of an integrated commercial system was made for two cases, a regional scale 50 million gallon (189,271 m3) per year facility (MGY) and a co-op scale 1&ndash, 2 MGY facility. The estimates for the minimum ethanol selling prices (MESP) depend heavily on the facility size and feedstock costs. For the 1&ndash, 2 MGY (3785&ndash, 7571 m3/y) facility, the MESP ranged from $5.61&ndash, $7.39 per gallon ($1.48&ndash, $1.95 per liter) for the four low-value feedstocks. These high costs suggest that the co-op scale even for the low-value feedstocks may not be economically sustainable. However, the MESP for the 50 MGY facility were significantly lower and comparable to gasoline prices ($2.24&ndash, $2.96 per gallon or $0.59&ndash, $0.78 per liter) for these low-value feedstocks, clearly showing the benefits of scale-up on construction costs and MESP.

A unique hybrid technology that uses renewable hydrogen (H 2 ) and carbon dioxide (CO 2 ) sequestered from large point sources, to produce fuels and chemicals has been proposed and tested. The primary objective of this research was to determine the feasibility of using two acetogenic bacteria to metabolize H 2 and CO 2 for the production of ethanol. Three experiments were conducted in small scale reactors to select a bacterium, feed gas composition and nutrient medium source to produce ethanol. The results indicated that Clostridium carboxidivorans produced 33% more ethanol and 66% less acetic acid compared to Clostridium ragsdalei , making C. carboxidivorans the better candidate for ethanol production. Furthermore, the removal of morpholinoethanesulfonic acid (MES) buffer from cotton seed extract (CSE) medium offered a low-cost medium for fermentations. Additionally, we observed that corn steep liquor (CSL) in the medium diversified the product range with both bacteria. Maximum concentrations of ethanol, n-butanol, n-hexanol, acetic acid, butyric acid, and hexanoic acid from different fermentation treatments were 2.78 g L −1 , 0.70 g L −1 , 0.52 g L −1 , 4.06 g L −1 , 0.13 g L −1 and 0.42 g L −1 , respectively. This study highlights the important role that acetogenic microbes can offer for CO 2 conversion into valuable fuels and chemicals.

This study investigates hydrolysis of cellulose from Eastern redcedar to glucose at high solids loading. Enzymatic hydrolysis of pretreated redcedar was performed with 0.5 ml Accelerase® 1500/g glucan (46 FPU/g glucan) using dry solids loading from 2% to 20% (w/w). Rheological challenges observed at high solids loading were overcome by adding stainless steel balls to shake flask reactors. The highest glucose concentration, 126 g/L (84% glucan-to-glucose yield), was obtained using 20% solids loading with stainless steel balls as a mixing aid. This enzymatic hydrolyzate was fermented into ethanol using Saccharomyces cerevisiae D 5 A to produce 52 g/L of ethanol (corresponding to 166 L/dry Mg of redcedar). Reducing enzyme dosage at 16% solids loading from 46 to 11.5 FPU/g glucan reduced glucan-to-glucose yields. This study has demonstrated the possibility of extracting sugars from the invasive species of Eastern redcedar with high solid loadings and their conversion into ethanol.

Large amounts of low-quality dates produced worldwide are wasted. Here, highly concentrated fructose syrups were produced via selective fermentation of date extracts with Saccharomyces cerevisiae. Syrups with 95.4-99.9 % (w/w) fructose yields were obtained from date extracts having an initial sugar range of 49-374 g/l without media supplementation; the corresponding ethanol yields were between 69 and 52 % (w/w). At 470 g initial sugars/l, fructose and ethanol yields were 84 and 47 % (w/w), respectively, and the product contained 62 % (w/w) fructose, which is higher than the widely available commercial 42 and 55 % (w/w) high fructose corn syrups. The commercial potential for conversion of waste dates to high-value products is thus demonstrated.

Anaerobic gut fungi represent a distinct early-branching fungal phylum (Neocallimastigomycota) and reside in the rumen, hindgut, and feces of ruminant and nonruminant herbivores. The genome of an anaerobic fungal isolate, Orpinomyces sp. strain C1A, was sequenced using a combination of Illumina and PacBio single-molecule real-time (SMRT) technologies. The large genome (100.95 Mb, 16,347 genes) displayed extremely low G+C content (17.0%), large noncoding intergenic regions (73.1%), proliferation of microsatellite repeats (4.9%), and multiple gene duplications. Comparative genomic analysis identified multiple genes and pathways that are absent in Dikarya genomes but present in early-branching fungal lineages and/or nonfungal Opisthokonta. These included genes for posttranslational fucosylation, the production of specific intramembrane proteases and extracellular protease inhibitors, the formation of a complete axoneme and intraflagellar trafficking machinery, and a near-complete focal adhesion machinery. Analysis of the lignocellulolytic machinery in the C1A genome revealed an extremely rich repertoire, with evidence of horizontal gene acquisition from multiple bacterial lineages. Experimental analysis indicated that strain C1A is a remarkable biomass degrader, capable of simultaneous saccharification and fermentation of the cellulosic and hemicellulosic fractions in multiple untreated grasses and crop residues examined, with the process significantly enhanced by mild pretreatments. This capability, acquired during its separate evolutionary trajectory in the rumen, along with its resilience and invasiveness compared to prokaryotic anaerobes, renders anaerobic fungi promising agents for consolidated bioprocessing schemes in biofuels production.

With the advancement of post-gasification processing, evaluating the effect of gas composition on ethanol production becomes important, as the composition could be shifted prior to fermentation to optimize ethanol yield. Increasing the overall efficiency of syngas conversion to bio-ethanol involves optimizing the use of the hydrogen (H2) component of syngas. This study focuses on the effect of different CO and H2 concentrations in the gaseous substrate on ethanol production and hydrogenase activity using Clostridium ragsdalei. Hydrogenase activity was observed to be the least inhibited by the lowest concentrations of H2 and CO. The maximum hydrogenase activity appeared early in the growth phase. At the lowest concentrations of CO and H2, hydrogenase appeared to recover activity slightly, and coincided with a depletion of acetic acid and an increase in ethanol concentrations. This phenomenon resulted in the highest concentration of ethanol observed (1.4 g L-1) at 10 days.

In this study, fructose and bioethanol were produced from date's extract by fermentation using Saccharomyces cerevisiae , mutant strain ATCC 36858 and wild strain STAR brand, where the latter was activated at different periods: 30 min, 1, 2 and 3 days. Profiles of sugar consumption and bioethanol production using STAR were found to be almost having the same pattern for all activation periods, while fructose was consumed as well as glucose. Enhancement of fructose in sugar can only be obtained at the expense of ethanol. In order to obtain 75 and 90% fructose in sugar, the respective losses in fructose exceeded 39 and 63%. On the other hand, the results demonstrate that S. cerevisiae ATCC 36858 could selectively convert glucose to ethanol and biomass with minimal fructose conversion. A high fructose yield above 91% of the original fructose was obtained with ATCC 36858. In addition, the ethanol yield was found to be 63% of the theoretical. Keywords: Saccharomyces cerevisiae , fructose, glucose, bioethanol, fermentation

Switchgrass ( Panicum virgatum ) was subjected to hydrothermolysis pretreatment and then used to study the effect of enzyme loading and temperature in a simultaneous saccharification and fermentation (SSF) with the thermotolerant yeast strain Kluyveromyces marxianus IMB3 at 8% solid loading. Various loadings of Accellerase 1500 between 0.1 and 1.1 mL g −1 glucan were tested in SSF at 45 °C (activity of enzyme was 82.2 FPU mL −1 ). The optimum enzyme loading was 0.7 mL g −1 glucan based on the six different enzyme loadings tested. SSFs were performed at 37, 41 and 45 °C with an enzyme loading of 0.7 mL g −1 glucan. The highest ethanol concentration of 22.5 g L −1 was obtained after 168 h with SSF at 45 °C, which was equivalent to 86% yield. Four different batch and fed-batch strategies were evaluated using a total solid loading of 12% (dry basis). About 32 g L −1 ethanol was produced with the four strategies, which was equivalent to 82% yield.

Gasification-fermentation is an emerging technology for the conversion of lignocellulosic materials into biofuels and specialty chemicals. For effective utilization of producer gas by fermenting bacteria, tar compounds produced in the gasification process are often removed by wet scrubbing techniques using acetone. In a preliminary study using biomass generated producer gas scrubbed with acetone, an accumulation of acetone and subsequent isopropanol production was observed. The effect of 2 g/L acetone concentrations in the fermentation media on growth and product distributions was studied with “Clostridium ragsdalei,” also known as Clostridium strain P11 or P11, and Clostridium carboxidivorans P7 or P7. The reduction of acetone to isopropanol was possible with “C. ragsdalei,” but not with P7. In P11 this reaction occurred rapidly when acetone was added in the acidogenic phase, but was 2.5 times slower when added in the solventogenic phase. Acetone at concentrations of 2 g/L did not affect the growth of P7, but ethanol increased by 41% and acetic acid concentrations decreased by 79%. In the fermentations using P11, growth was unaffected and ethanol concentrations increased by 55% when acetone was added in the acidogenic phase. Acetic acid concentrations increased by 19% in both the treatments where acetone was added. Our observations indicate that P11 has a secondary alcohol dehydrogenase that enables it to reduce acetone to isopropanol, while P7 lacks this enzyme. P11 offers an opportunity for biological production of isopropanol from acetone reduction in the presence of gaseous substrates (CO, CO2, and H2). Biotechnol. Bioeng. 2011;108: 2330–2338. © 2011 Wiley Periodicals, Inc.

The effect of dithiothreitol (DTT) on enhancing ethanol production from syngas using Clostridium strain P11 was investigated in 250-mL serum bottles. Reducing agents help in regeneration of NADH from NAD+. NADH is utilized in the production of alcohol from aldehydes. Strain P11 was fed with syngas every 24 h and samples were collected to measure pH, cell mass and product concentrations. Various concentrations of DTT were examined. Results showed over fourfold increase in ethanol production in media that contained at least 7.5 g/L of DTT after 360 h of fermentation compared to the control medium (without DTT). Similar pH, cell mass and acetic acid profiles were observed in the control and DTT media. The results suggested that the use of small concentrations of DTT in the broth enhances ethanol production from syngas, which is an important factor towards making gasification-fermentation technology an economically viable process.

A highly reliable experimental system that consistently closed the overall water balance to within 5% was developed to study the role of a microporous layer (MPL), attached to carbon paper porous transport layer (PTL), on the water transport and performance of a standard 100 cm2 active area PEM fuel cell. Various combinations of cells were built and tested with PTLs at the electrodes using either carbon fibre paper with a MPL (SGL 10BB) or carbon fibre paper without a MPL (SGL 10BA). The net water drag coefficient at three current densities (0.3, 0.5 and 0.7 A cm−2) for two combinations of anode/cathode relative humidity (60/100% and 100/60%) and stoichiometric ratios of H2/air (1.4/3 and 1.4/2) was determined from water balance measurements. The addition of a MPL to the carbon fibre paper PTL at the cathode did not cause a statistically significant change to the overall drag coefficient although there was a significant improvement to the fuel cell performance and durability when a MPL was used at the cathode. The presence of a MPL on either electrode or on both electrodes also exhibited similar performance compared to when the MPL was placed at the cathode. These results indicate that the presence of MPL indeed improves the cell performance although it does not affect the net water drag coefficient. The correlation between cell performance and global water transport cannot be ascertained and warrants further experimental investigation.

To assist in the practical implementation of irreversible (reactive) hydrogen storage for automotive applications, the separation of the reactants and products is beneficial. This separation could be effective on board where the build-up of reaction products might prove detrimental to operation, or in electrochemical regeneration of the reactant. Three commercial nanofiltration (NF) membranes (FilmTecTM NF-270, FilmTecTM NF-70 and GE Osmonics thin film (TF), NF-DL) were tested at different operating conditions for the separation of sodium metaborate tetrahydrate ( NaBO 2 · 4 H 2 O ) from sodium borohydride ( NaBH 4 ) for hydrogen storage applications. FilmTecTM NF-270 and GE Osmonics TF, NF-DL membranes successfully separated NaBO 2 · 4 H 2 O from NaBH 4 . The separation performance and chemical stability of the FilmTecTM NF-270 membrane were higher than for other tested membranes. The rejection of NaBO 2 · 4 H 2 O by the FilmTecTM NF-270 membrane and flux rate were 76% and 17.33 L m 2 / h , respectively, at 350 psig and pH 12.4 for a feed that contained 90 g/L NaBH 4 and 62 g/L NaBO 2 · 4 H 2 O . The other two membranes failed to reject NaBO 2 · 4 H 2 O at the above operating conditions. However, NaBO 2 · 4 H 2 O rejection by the GE Osmonics TF, NF-DL membrane was 40% when the feed contained 60 and 30 g/L of NaBH 4 and NaBO 2 · 4 H 2 O , respectively. This separation could be potentially applied to increase the concentration of NaBH 4 in NaOH stabilized solutions and in the recycling of NaBO 2 · 4 H 2 O .

Clostridium carboxidivorans was grown on model syngas (CO:H2:CO2 [70:20:10]) in a defined nutrient medium with concentrations of nitrogen, phosphate and trace metals formulated to enhance production of higher alcohols. C. carboxidivorans was successfully grown in a limited defined medium (no yeast extract, no MES buffer and minimal complex chemical inputs) using an improved fermentation protocol. Low partial pressure of CO in the headspace, coupled with restricted mass transfer for CO and H2, was required for successful fermentation. In the absence of substrate inhibition (particularly from CO), growth limitation increased production of alcohols, especially butanol and hexanol. Concentrations of butanol (over 1.0g/L), hexanol (up to 1.0g/L) and ethanol (over 3.0g/L) were achieved in bottle fermentations. Minimal medium and controlled supply of CO and H2 should be used in characterizing candidate butanol and hexanol producing strains to select for commercial potential.

A test station was modified to collect water from the anode and cathode exhaust streams of a PEM fuel cell. Various combinations of cells were built and tested with (SGL10BA) and without microporous layers (SGL10BB) on the electrodes. Cells with a MPL at the cathode performed much better compared to cells with no MPL and the degradation rates were decreased. Water balance experiments were conducted with two combinations of inlet relative humidities (RH): 100% anode-60% cathode and 60% anode-100% cathode at current densities of 300, 500 and 700 mA/cm2. For the range of current densities studied, the net drag coefficient at 60{degree sign}C was -0.08-+0.08 mol H2O/mol H+ for RH 60% anode and 0.01-0.2 mol H2O/mol H+ for RH 100% anode. There was no significant change in the drag coefficient attributable to the presence of the MPL on the cathode. A MPL on the anode did affect the net water drag.