Effects of Forage Quality and Cell Wall Constituents of Bermuda Grass on Biochemical Conversion to Ethanol

Bermuda grass is an attractive candidate as a feedstock for biofuel production because over four million hectares of Bermuda grass are already grown for forage in the Southern USA. Because both rumen digestion and biochemical conversion to ethanol depend upon enzymatic conversion of the cell wall polysaccharides into fermentable sugars, it is probable that grasses bred for increased forage quality would be more amenable for ethanol production. However, it is not known how variation in rumen digestibility and cell wall/fiber components correlates with efficiency of conversion to ethanol via fermentation. The objective of this research was to determine relationships between ethanol production evaluated by simultaneous saccharification and fermentation (SSF), 72-h in vitro ruminal dry matter digestibility (IVDMD), in vitro ruminal gas production after 24 and 96 h, and biomass composition for 50 genetically diverse Bermuda grass accessions. The Bermuda grass samples were subjected to standard 72-h IVDMD and forage fiber analyses. Also, in separate labs, gas production was measured in sealed volume-calibrated vials after 24 (NNG24) and 96 h (NNG96) of in vitro fermentation by ruminal fluid; ethanol and pentose sugar productions were measured from a bench-top SSF procedure; cell wall constituents were determined by the Uppsala Dietary Fiber Method; and total nitrogen, carbon, and ash concentrations were determined by using the LECO combustion method. Ethanol production was moderately correlated with IVDMD (r?=?0.55) and NNG96 (r?=?0.63) but highly correlated with NNG24 (r?=?0.93). Ethanol was negatively correlated with neutral detergent fiber (NDF; r?=??0.53) and pentose sugars (r?=??0.60), but not correlated with glucose content. Regression models indicated that NDF and cell wall pentose sugar concentrations had significant negative effects on ethanol production. Variation among entries for IVDMD was affected by variability of NDF, pentose sugar concentrations, and biomass nitrogen content. Variation in Klason lignin content had only minor negative impacts on ethanol production and IVDMD. Biochemical conversion efficiency of Bermuda grass by SSF can be best estimated by NNG24 but not by IVDMD.     

Trends in biotechnological production of fuel ethanol from different feedstocks

Present work deals with the biotechnological production of fuel ethanol from different raw materials. The different technologies for producing fuel ethanol from sucrose-containing feedstocks (mainly sugar cane), starchy materials and lignocellulosic biomass are described along with the major research trends for improving them. The complexity of the biomass processing is recognized through the analysis of the different stages involved in the conversion of lignocellulosic complex into fermentable sugars. The features of fermentation processes for the three groups of studied feedstocks are discussed. Comparative indexes for the three major types of feedstocks for fuel ethanol production are presented. Finally, some concluding considerations on current research and future tendencies in the production of fuel ethanol regarding the pretreatment and biological conversion of the feedstocks are presented.     

Fusarium oxysporum: status in bioethanol production

Fermentation of lignocellulosic materials to ethanol and other solvents provides an alternative way of treating wastes and producing chemical feedstocks and fuel additives. Considerable efforts have been made in past 10 years to improve the process based on lignocellulosic biomass and hydrolysate that contains a complex mixture of sugars, decomposition products of sugars, and sometimes the inhibitory levels of soluble lignin. Despite the relative abundance of D-xylose in crop and forest residues it has not been found efficiently fermentable by most of the microorganisms. Recent research has revealed that D-xylose may be fermented to ethanol and organic acids. Recently, several strains of Fusarium oxysporum have been found to have potential for converting not only D-xylose, but also cellulose to ethanol in a one-step process. Distinguishing features of F. oxysporum for ethanol production in comparison to other organisms are identified. These include the advantage of in situ cellulase production and cellulose fermentation, pentose fermentation, and the tolerance of sugars and ethanol. The main disadvantage is the slow conversion rate when compared with yeast.     

Dedicated Herbaceous Biomass Feedstock Genetics and Development

Biofuels and bio-based products can be produced from a wide variety of herbaceous feedstocks. To supply enough biomass to meet the needs of a new bio-based economy, a suite of dedicated biomass species must be developed to accommodate a range of growing environments throughout the USA. Researchers from the US Department of Agriculture’s Agricultural Research Service (USDA-ARS) and collaborators associated with the USDA Regional Biomass Research Centers have made major progress in understanding the genetics of switchgrass, sorghum, and other grass species and have begun to use this knowledge to develop new cultivars with high yields and appropriate traits for efficient conversion to bio-based products. Plant geneticists and breeders have discovered genes that reduce recalcitrance for biochemical conversion to ethanol and drop-in fuels. Progress has also been made in finding genes that improve production under biotic and abiotic stress from diseases, pests, and climatic variations.     

Ethanol fermentation from biomass resources: current state and prospects

In recent years, growing attention has been devoted to the conversion of biomass into fuel ethanol, considered the cleanest liquid fuel alternative to fossil fuels. Significant advances have been made towards the technology of ethanol fermentation. This review provides practical examples and gives a broad overview of the current status of ethanol fermentation including biomass resources, microorganisms, and technology. Also, the promising prospects of ethanol fermentation are especially introduced. The prospects included are fermentation technology converting xylose to ethanol, cellulase enzyme utilized in the hydrolysis of lignocellulosic materials, immobilization of the microorganism in large systems, simultaneous saccharification and fermentation, and sugar conversion into ethanol.     

Xylose isomerase improves growth and ethanol production rates from biomass sugars for both Saccharomyces pastorianus and Saccharomyces cerevisiae

The demand for biofuel ethanol made from clean, renewable nonfood sources is growing. Cellulosic biomass, such as switch grass (Panicum virgatum L.), is an alternative feedstock for ethanol production; however, cellulosic feedstock hydrolysates contain high levels of xylose, which needs to be converted to ethanol to meet economic feasibility. In this study, the effects of xylose isomerase on cell growth and ethanol production from biomass sugars representative of switch grass were investigated using low cell density cultures. The lager yeast species Saccharomyces pastorianus was grown with immobilized xylose isomerase in the fermentation step to determine the impact of the glucose and xylose concentrations on the ethanol production rates. Ethanol production rates were improved due to xylose isomerase; however, the positive effect was not due solely to the conversion of xylose to xylulose. Xylose isomerase also has glucose isomerase activity, so to better understand the impact of the xylose isomerase on S. pastorianus, growth and ethanol production were examined in cultures provided fructose as the sole carbon. It was observed that growth and ethanol production rates were higher for the fructose cultures with xylose isomerase even in the absence of xylose. To determine whether the positive effects of xylose isomerase extended to other yeast species, a side-by-side comparison of S. pastorianus and Saccharomyces cerevisiae was conducted. These comparisons demonstrated that the xylose isomerase increased ethanol productivity for both the yeast species by increasing the glucose consumption rate. These results suggest that xylose isomerase can contribute to improved ethanol productivity, even without significant xylose conversion.     

微生物木糖发酵产乙醇的代谢工程

利用木质纤维素发酵生产乙醇具有广泛的应用前景。而自然界中缺少有效转化木糖为乙醇的微生物是充分利用纤维素水解产物、提高乙醇产率、降低生产成本的关键因素。多年来研究者利用分子生物学技术对微生物菌株进行了代谢工程改造,使其能更有效地利用木糖生产乙醇。以下主要对运动发酵单胞菌、大肠杆菌和酵母等候选产乙醇微生物的木糖代谢工程研究进展进行了概述。     

Weedy lignocellulosic feedstock and microbial metabolic engineering: advancing the generation of ‘Biofuel’

Lignocellulosic materials are the most abundant renewable organic resources (~200 billion tons annually) on earth that are readily available for conversion to ethanol and other value-added products, but they have not yet been tapped for the commercial production of fuel ethanol. The lignocellulosic substrates include woody substrates such as hardwood (birch and aspen, etc.) and softwood (spruce and pine, etc.), agro residues (wheat straw, sugarcane bagasse, corn stover, etc.), dedicated energy crops (switch grass, and Miscanthus etc.), weedy materials (Eicchornia crassipes, Lantana camara etc.), and municipal solid waste (food and kitchen waste, etc.). Despite the success achieved in the laboratory, there are limitations to success with lignocellulosic substrates on a commercial scale. The future of lignocellulosics is expected to lie in improvements of plant biomass, metabolic engineering of ethanol, and cellulolytic enzyme-producing microorganisms, fullest exploitation of weed materials, and process integration of the individual steps involved in bioethanol production. Issues related to the chemical composition of various weedy raw substrates for bioethanol formation, including chemical composition-based structural hydrolysis of the substrate, need special attention. This area could be opened up further by exploring genetically modified metabolic engineering routes in weedy materials and in biocatalysts that would make the production of bioethanol more efficient.     

A Survey of Bioenergy Research in Forest Service Research and Development

Forest biomass represents 25–30 % of the annual biomass available in the USA for conversion into bio-based fuels, bio-based chemicals, and bioproducts in general. The USDA Forest Service Research and Development (R&D) has been focused on producing products from forest biomass since its inception in 1905, with direct combustion, solid sawn lumber, pulp and paper, ethanol as fuel, and silvichemicals all among the mission areas of product research and development. The renewed national interest in biomass conversion to fuels and chemicals is supportive of the most critical need of USDA Forest Service R&D, uses for small-diameter trees and other forest biomass that needs to be removed in the fuel mitigation–fire suppression and forest restoration work of the USDA Forest Service. This paper will summarize the recent USDA Forest Service research on direct combustion, fuel pellets, and conversion of forest biomass to ethanol, both as stand-alone biorefinery processes and as an addition to the traditional wood pulping process.     

A new dawn for industrial photosynthesis

Several emerging technologies are aiming to meet renewable fuel standards, mitigate greenhouse gas emissions, and provide viable alternatives to fossil fuels. Direct conversion of solar energy into fungible liquid fuel is a particularly attractive option, though conversion of that energy on an industrial scale depends on the efficiency of its capture and conversion. Large-scale programs have been undertaken in the recent past that used solar energy to grow innately oil-producing algae for biomass processing to biodiesel fuel. These efforts were ultimately deemed to be uneconomical because the costs of culturing, harvesting, and processing of algal biomass were not balanced by the process efficiencies for solar photon capture and conversion. This analysis addresses solar capture and conversion efficiencies and introduces a unique systems approach, enabled by advances in strain engineering, photobioreactor design, and a process that contradicts prejudicial opinions about the viability of industrial photosynthesis. We calculate efficiencies for this direct, continuous solar process based on common boundary conditions, empirical measurements and validated assumptions wherein genetically engineered cyanobacteria convert industrially sourced, high-concentration CO₂ into secreted, fungible hydrocarbon products in a continuous process. These innovations are projected to operate at areal productivities far exceeding those based on accumulation and refining of plant or algal biomass or on prior assumptions of photosynthetic productivity. This concept, currently enabled for production of ethanol and alkane diesel fuel molecules, and operating at pilot scale, establishes a new paradigm for high productivity manufacturing of nonfossil-derived fuels and chemicals.     

Low temperature pre-treatment of domestic sewage in an anaerobic hybrid or an anaerobic filter reactor

The ability of pigs to use nitrogen and energy in Bermuda grass was evaluated in order to assess whether Bermuda grass harvested from spray fields could be fed to pigs as a means to recycle nitrogen. Digestibility of Bermuda grass incorporated into corn-soybean meal diets was evaluated in heavy finishing pigs and gestating sows. Results suggest that Bermuda grass digestibility is negative in animals not adapted to a high-fiber diet. Enzymes improve this digestibility, but even with enzymes, nitrogen digestibility was poor. Pigs fed a diet containing 10% Bermuda grass required a one week adaptation period for maximal digestion; following adaptation, pigs can digest approximately 40% of the energy in Bermuda grass but none of the nitrogen. Feeding Bermuda grass to pigs as a means of recycling nitrogen is thus not recommended.     

Potential for Production of Perennial Biofuel Feedstocks in Conservation Buffers on the Coastal Plain of Georgia,USA

With global increases in the production of cellulosic biomass for fuel, or “biofuel,” concerns over potential negative effects of using land for biofuel production have promoted attention to concepts of agricultural landscape design that sustainably balance tradeoffs between food, fuel, fiber, and conservation. The Energy Independence Security Act (EISA) of 2007 mandates an increase in advanced biofuels to 21 billion gallons in 2022. The southeastern region of the USA has been identified as a contributor to meeting half of this goal. We used a GIS-based approach to estimate the production and N-removal potential of three perennial biofeedstocks planted as conservation buffers (field borders associated with riparian buffers, and grassed waterways) on the Coastal Plain of Georgia, USA. Land cover, hydrology, elevation, and soils data were used to identify locations within agricultural landscapes that are most susceptible to runoff, erosion, and nutrient loss. We estimated potential annual biomass production from these areas to be: 2.5–3.5 Tg for giant miscanthus (Miscanthus?×?giganteus), 2–8.6 Tg for “Merkeron” napier grass (Pennisetum purpureum), and 1.9–7.5 Tg for “Alamo” switchgrass (Panicum virgatum). When production strategies were taken into consideration, we estimated total biomass yield of perennial grasses for the Georgia Coastal Plain at 2.2–9.4 Tg year^?1. Using published rates of N removal and ethanol conversion, we calculated the amount of potential N removal by these systems as 8100–51,000 Mg year^?1 and ethanol fuel production as 778–3296 Ml year^?1 (206 to 871 million gal. US).     

Metabolic engineering of bacteria for ethanol production

Technologies are available which will allow the conversion of lignocellulose into fuel ethanol using genetically engineered bacteria. Assembling these into a cost-effective process remains a challenge. Our work has focused primarily on the genetic engineering of enteric bacteria using a portable ethanol production pathway. Genes encoding Zymomonas mobilis pyruvate decarboxylase and alcohol dehydrogenase have been integrated into the chromosome of Escherichia coli B to produce strain KO11 for the fermentation of hemicellulose-derived syrups. This organism can efficiently ferment all hexose and pentose sugars present in the polymers of hemicellulose. Klebsiella oxytoca M5A1 has been genetically engineered in a similar manner to produce strain P2 for ethanol production from cellulose. This organism has the native ability to ferment cellobiose and cellotriose, eliminating the need for one class of cellulase enzymes. The optimal pH for cellulose fermentation with this organism (pH 5.0-5.5) is near that of fungal cellulases. The general approach for the genetic engineering of new biocatalysts has been most successful with enteric bacteria thus far. However, this approach may also prove useful with Gram-positive bacteria which have other important traits for lignocellulose conversion. Many opportunities remain for further improvements in the biomass to ethanol processes. These include the development of enzyme-based systems which eliminate the need for dilute acid hydrolysis or other pretreatments, improvements in existing pretreatments for enzymatic hydrolysis, process improvements to increase the effective use of cellulase and hemicellulase enzymes, improvements in rates of ethanol production, decreased nutrient costs, increases in ethanol concentrations achieved in biomass beers, increased resistance of the biocatalysts to lignocellulosic-derived toxins, etc. To be useful, each of these improvements must result in a decrease in the cost for ethanol production. Copyright 1998 John Wiley & Sons, Inc.     

Functional group and fertilization affect the composition and bioenergy yields of prairie plants

Prairies used for bioenergy production have potential to generate marketable products while enhancing environmental quality, but little is known about how prairie species composition and nutrient management affect the suitability of prairie biomass for bioenergy production. We determined how functional‐group identity and nitrogen fertilization affected feedstock characteristics and estimated bioenergy yields of prairie plants, and compared those prairie characteristics to that of corn stover. We tested our objectives with a field experiment that was set up as a 5 × 2 incomplete factorial design with C₃ grasses, C₄ grasses, legumes, and multi‐functional‐group mixtures grown with and without nitrogen fertilizer; a fertilized corn treatment was also included. We determined cell wall, hemicellulose, cellulose, and ash concentrations; ethanol conversion ratios; gross caloric ratios; aboveground biomass production; ethanol yields; and energy yields for all treatments. Prairie functional‐group identity affected the biomass feedstock characteristics, whereas nitrogen fertilization did not. Functional group and fertilization had a strong effect on aboveground biomass production, which was the major predictor of ethanol and energy yields. C₄ grasses, especially when fertilized, had among the most favorable bioenergy characteristics with high estimated ethanol conversion ratios and nongrain biomass production, relatively high gross caloric ratios, and low ash concentrations. The bioenergy characteristics of corn stover, from an annual C₄ grass, were similar to those of the biomass of perennial C₄ grasses. Both functional‐group composition and nitrogen fertility management were found to be important in optimizing bioenergy production from prairies.     

木质纤维生产燃料乙醇工艺的研究进展

利用丰富而廉价的木质纤维原料代替粮食生产燃料乙醇,对经济和社会的可持续发展有着重要的意义。以木质纤维为原料发酵生产燃料乙醇可分为4种工艺:分步糖水解化发酵法、同步糖化发酵法、同步糖化共发酵法和直接微生物转化法。介绍了以上4种工艺的研究进展,并对今后进一步研究提出了建议。     

Meeting US biofuel goals with less land: the potential of Miscanthus

Biofuels from crops are emerging as a Jekyll & Hyde – promoted by some as a means to offset fossil fuel emissions, denigrated by others as lacking sustainability and taking land from food crops. It is frequently asserted that plants convert only 0.1% of solar energy into biomass, therefore requiring unacceptable amounts of land for production of fuel feedstocks. The C₄ perennial grass Miscanthus × giganteus has proved a promising biomass crop in Europe, while switchgrass ( Panicum virgatum ) has been tested at several locations in N. America. Here, replicated side-by-side trials of these two crops were established for the first time along a latitudinal gradient in Illinois. Over 3 years of trials, Miscanthus × giganteus achieved average annual conversion efficiencies into harvestable biomass of 1.0% (30 t ha⁻¹) and a maximum of 2.0% (61 t ha⁻¹), with minimal agricultural inputs. The regionally adapted switchgrass variety Cave-in-Rock achieved somewhat lower yields, averaging 10 t ha⁻¹. Given that there has been little attempt to improve the agronomy and genetics of these grasses compared with the major grain crops, these efficiencies are the minimum of what may be achieved. At this 1.0% efficiency, 12 million hectares, or 9.3% of current US cropland, would be sufficient to provide 133 × 10⁹ L of ethanol, enough to offset one-fifth of the current US gasoline use. In contrast, maize grain from the same area of land would only provide 49 × 10⁹ L, while requiring much higher nitrogen and fossil energy inputs in its cultivation.     

Applications of yeast cell-surface display in bio-refinery

The dependency on depleting natural resources is a challenge for energy security that can be potentially answered by bioenergy. Bioenergy is derived from starchy and lignocellulosic biomass in the form of bioethanol or from vegetable oils in the form of biodiesel fuel. The acid and enzymatic methods have been developed for the hydrolysis of biomass and for transesterifiaction of plant oils. However, acid hydrolysis results in the production of unnatural compounds which has adverse effects on yeast fermentation. Recent advancements in the yeast cell surface engineering developed strategies to genetically immobilize amylolytic, cellulolytic and xylanolytic enzymes on yeast cell surface for the production of fuel ethanol from biomass. This review gives an insight in to the recent technological developments in the production of bioenergy, i.e, bioethanol using surface engineered yeast.     

Hydrolysis of lignocellulosic materials for ethanol production: a review

Lignocellulosic biomass can be utilized to produce ethanol, a promising alternative energy source for the limited crude oil. There are mainly two processes involved in the conversion: hydrolysis of cellulose in the lignocellulosic biomass to produce reducing sugars, and fermentation of the sugars to ethanol. The cost of ethanol production from lignocellulosic materials is relatively high based on current technologies, and the main challenges are the low yield and high cost of the hydrolysis process. Considerable research efforts have been made to improve the hydrolysis of lignocellulosic materials. Pretreatment of lignocellulosic materials to remove lignin and hemicellulose can significantly enhance the hydrolysis of cellulose. Optimization of the cellulase enzymes and the enzyme loading can also improve the hydrolysis. Simultaneous saccharification and fermentation effectively removes glucose, which is an inhibitor to cellulase activity, thus increasing the yield and rate of cellulose hydrolysis.     

Agave for tequila and biofuels: an economic assessment and potential opportunities

This paper explores the economic viability of producing biofuels from Agave in Mexico and the potential for it to complement the production of tequila or mescal. We focus on Agave varieties currently being used by the tequila industry to produce two beverages, tequila and mescal, and explore the potential for biofuel production from these plants. Without competing directly with beverage production, we discuss the economic costs and benefits of converting Agave by‐products to liquid fuel as an additional value‐added product and expanding cultivation of Agave on available land. We find that the feedstock cost for biofuel from the Agave piña alone could be more than US$3 L^?1 on average. This is considerably higher than the feedstock costs of corn ethanol and sugarcane ethanol. However, there may be potential to reduce these costs with higher conversion efficiencies or by using sugar present in other parts of the plant. The costs of cellulosic biofuels using the biomass from the entire plant could be lower depending on the conversion efficiency of biomass to fuel and the additional costs of harvesting, collecting and transporting that biomass.     

Global Potential of Rice Husk as a Renewable Feedstock for Ethanol Biofuel Production

The production of ethanol for the energy market has traditionally been from corn and sugar cane biomass. The use of such biomass as energy feedstocks has recently been criticised as ill-fated due to competitive threat against food supplies. At the same time, ethanol production from cellulosic biomass is becoming increasingly popular. In this paper, we analyse rice husk (RH) as a cellulosic feedstock for ethanol biofuel production on the ground of its abundance. The global potential production of bioethanol from RH is estimated herein and found to be in the order of 20.9 to 24.3 GL per annum, potentially satisfying around one fifth of the global ethanol biofuel demand for a 10% gasohol fuel blend. Furthermore, we show that this is especially advantageous for Asia, in particular, India and China, where economic growth and demand for energy are exploding.