Integrated Economic and Environmental Assessment of Cellulosic Biofuel Production in an Agricultural Watershed

SWAT watershed model simulated biomass yield and pollutant loadings were integrated with associated economic costs of farm production and transport to study two dedicated energy crops, switchgrass and Miscanthus, and corn stover, as feedstocks for a cellulosic biorefinery. A multi-level spatial optimization (MLSOPT) framework was employed to get spatially explicit cropping plans for a watershed under the assumption that the watershed supplies biomass to a hypothetical biorefinery considering both the biochemical and the thermochemical conversion pathways. Consistent with previous studies, the perennial grasses had higher biomass yield than corn stover, with considerably lower sediment, nitrogen, and phosphorus loadings, but their costs were higher. New insights were related to the tradeoffs between cost, feedstock production, and the level and form of environmental quality society faces as it implements the Renewable Fuel Standard. Economically, this involved calculating the farthest distance a biorefinery would be willing to drive to source corn residue before procuring a single unit of perennial grasses from productive agricultural soils.     

Prospects for Cellulosic Biofuel Production in the Northeastern United States: A Scenario Analysis

Secure access to energy and food are two of the challenges facing the Northeast region of the United States. Traditional biofuel feedstocks, such as corn and oil seed, are able to satisfy energy requirements. However, they compete with food production for desirable land and water resources and, in any case, are not likely to exploit the region's current comparative advantages. This study investigates a potential solution to the energy security problem in the Northeast: biofuel from advanced feedstock in the form of net forest growth and woody wastes, of which the region has abundant endowments. The federal government has committed to requiring 79.5 billion liters (BL) of advanced biofuel production annually by 2022. We evaluate both the physical capacity for its production and its cost competitiveness using an input‐output model of consumption, production, and trade in the 13‐state region. The model minimizes resource use required to satisfy given consumer demand using alternative technological options and subject to resource constraints. We compile data from the technical literature quantifying state‐level biofuel feedstock endowments and the technological requirements for cellulosic ethanol production. We find that exploiting the region's endowment of cellulosic feedstock requires either making the price of biofuels competitive with gasoline through subsidies or restricting imports of gasoline. Based on this initial investigation, we conclude that the region can produce significant amounts of advanced biofuel, up to 20.28 BL of cellulosic ethanol per year, which could displace nearly 12.5% of the gasoline that is now devoted to motorized transport in the region.     

Development of Sustainable Corn Stover Harvest Strategies for Cellulosic Ethanol Production

To prepare for a 2014 launch of commercial scale cellulosic ethanol production from corn/maize (Zea mays L.) stover, POET-DSM near Emmetsburg, IA has been working with farmers, researchers, and equipment dealers through “Project Liberty” on harvest, transportation, and storage logistics of corn stover for the past several years. Our objective was to evaluate seven stover harvest strategies within a 50-ha (125 acres) site on very deep, moderately well to poorly drained Mollisols, developed in calcareous glacial till. The treatments included the following: conventional grain harvest (no stover harvest), grain plus a second-pass rake and bale stover harvest, and single-pass grain plus cob-only biomass, grain plus vegetative material other than grain [(MOG) consisting of cobs, husks, and upper plant parts], grain plus all vegetative material from the ear shank upward (high cut), and all vegetative material above a 10 cm stubble height (low cut), with a John Deere 9750 STS combine, and grain plus direct baling of MOG with an AgCo harvesting system. Average grain yields were 11.4, 10.1, 9.7, and 9.5 Mg ha^?1 for 2008, 2009, 2010, and 2011, respectively. Average stover harvest ranged from 0 to 5.6 Mg ha^?1 and increased N, P, and K removal by an average of 11, 1.6, and 15 kg Mg^?1, respectively. Grain yield in 2009 showed a significant positive response to higher 2008 stover removal rates, but grain yield was not increased in 2010 or 2011 due to prior-year stover harvest. High field losses caused the direct-bale treatment to have significantly lower grain yield in 2011 because the AgCo system could not pick up the severely lodged crop. We conclude that decreases in grain yield across the 4 years were due more to seasonal weather patterns, spatial variability, and not rotating crops than to stover harvest.     

Production of Bacterial Proteins from Cellulosic Materials

An aerobic mesophilic Pseudomonas sp. isolated from activated sludge degraded different cellulosic materials to varying-degrees. The degradability was mostly influenced by the lignin content and the crystalline nature of the substrate. Filter paper and cotton fibres, containing little lignin, were degraded maximally. Lignin-rich Pinus and Larix needles were digested to a lesser extent. There was a difference between natural substrates and substrates that had undergone industrial treatment. At maximal protein production, the isolate converted 11–40% (w/w) of cellulosic substrates into proteins. Alkali treatment of the substrates had only little effect. Cellulose contained in pig-waste was also degraded by the isolate. At first there was a period of protein production, but after long incubation, the efficiency of conversion of cellulose to protein decreased.     

Generating Phenotypic Diversity in a Fungal Biocatalyst to Investigate Alcohol Stress Tolerance Encountered during Microbial Cellulosic Biofuel Production

Consolidated bioprocessing (CBP) of lignocellulosic biomass offers an alternative route to renewable energy. The crop pathogen Fusarium oxysporum is a promising fungal biocatalyst because of its broad host range and innate ability to co-saccharify and ferment lignocellulose to bioethanol. A major challenge for cellulolytic CBP-enabling microbes is alcohol inhibition. This research tested the hypothesis that Agrobacterium tumefaciens - mediated transformation (ATMT) could be exploited as a tool to generate phenotypic diversity in F. oxysporum to investigate alcohol stress tolerance encountered during CBP. A random mutagenesis library of gene disruption transformants (n=1,563) was constructed and screened for alcohol tolerance in order to isolate alcohol sensitive or tolerant phenotypes. Following three rounds of screening, exposure of select transformants to 6% ethanol and 0.75% n-butanol resulted respectively in increased (≥11.74%) and decreased (≤43.01%) growth compared to the wild –type (WT). Principal component analysis (PCA) quantified the level of phenotypic diversity across the population of genetically transformed individuals and isolated candidate strains for analysis. Characterisation of one strain, Tr. 259, ascertained a reduced growth phenotype under alcohol stress relative to WT and indicated the disruption of a coding region homologous to a putative sugar transporter (FOXG_09625). Quantitative PCR (RT-PCR) showed FOXG_09625 was differentially expressed in Tr. 259 compared to WT during alcohol-induced stress (P<0.05). Phylogenetic analysis of putative sugar transporters suggests diverse functional roles in F. oxysporum and other filamentous fungi compared to yeast for which sugar transporters form part of a relatively conserved family. This study has confirmed the potential of ATMT coupled with a phenotypic screening program to select for genetic variation induced in response to alcohol stress. This research represents a first step in the investigation of alcohol tolerance in F. oxysporum and has resulted in the identification of several novel strains, which will be of benefit to future biofuel research.     

Streamflow Impacts of Biofuel Policy-Driven Landscape Change

Likely changes in precipitation (P) and potential evapotranspiration (PET) resulting from policy-driven expansion of bioenergy crops in the United States are shown to create significant changes in streamflow volumes and increase water stress in the High Plains. Regional climate simulations for current and biofuel cropping system scenarios are evaluated using the same atmospheric forcing data over the period 1979–2004 using the Weather Research Forecast (WRF) model coupled to the NOAH land surface model. PET is projected to increase under the biofuel crop production scenario. The magnitude of the mean annual increase in PET is larger than the inter-annual variability of change in PET, indicating that PET increase is a forced response to the biofuel cropping system land use. Across the conterminous U.S., the change in mean streamflow volume under the biofuel scenario is estimated to range from negative 56% to positive 20% relative to a business-as-usual baseline scenario. In Kansas and Oklahoma, annual streamflow volume is reduced by an average of 20%, and this reduction in streamflow volume is due primarily to increased PET. Predicted increase in mean annual P under the biofuel crop production scenario is lower than its inter-annual variability, indicating that additional simulations would be necessary to determine conclusively whether predicted change in P is a response to biofuel crop production. Although estimated changes in streamflow volume include the influence of P change, sensitivity results show that PET change is the significantly dominant factor causing streamflow change. Higher PET and lower streamflow due to biofuel feedstock production are likely to increase water stress in the High Plains. When pursuing sustainable biofuels policy, decision-makers should consider the impacts of feedstock production on water scarcity.     

Current and Potential Biofuel Production from Plant Oils

Environmental concerns and depletion of fossil fuels along with government policies have led to the search for alternative fuels from various renewable and sustainable feedstocks. This review provides a critical overview of the chemical composition of common commercial plant oils, i.e., palm oil, olive oil, rapeseed oil, castor oil, WCO, and CTO and their recent trends toward potential biofuel production. Plant oils with a high energy content are primarily composed of triglycerides (generally >?95%), accompanied by diglycerides, monoglycerides, and free fatty acids. The heat content of plant oils is close to 90% for diesel fuels. The oxygen content is the most important difference in chemical composition between fossil oils and plant oils. Triglycerides can even be used directly in diesel engines. However, their high viscosity, low volatility, and poor cold flow properties can lead to engine problems. These problems require that plant oils need to be upgraded if they are to be used as a fuel in conventional diesel engines. Biodiesel, biooil, and renewable diesel are the three major biofuels obtained from plant oils. The main constraint associated with the production of biodiesel is the cost and sustainability of the feedstock. The renewable diesel obtained from crude tall oil is more sustainable than biofuels obtained from other feedstocks. The fuel properties of renewable diesel are similar to those of fossil fuels with reduced greenhouse gas emissions. In this review, the chemical composition of common commercial plant oils, i.e., palm oil, olive oil, rapeseed oil, castor oil, and tall oil, are presented. Both their major and minor components are discussed. Their compositions and fuel properties are compared to both fossil fuels and biofuels.     

Biomass and Cellulosic Ethanol Production of Forage Sorghum Under Limited Water Conditions

This study presents results from a 2-year evaluation of biomass and cellulosic ethanol (EtOH) production potential of forage sorghum (Sorghum bicolor L. Moench) cultivars differing in brown midrib trait (i.e., bmr12) under dryland (no irrigation) and limited irrigation (2.88 mm?day^?1; subsurface drip) in the semiarid Southern High Plains of the USA. Commercial cultivar Sorghum Partners 1990 (SP 1990, conventional non-bmr) produced significantly more biomass (29–62 %) than a bmr12 cultivar PaceSetter bmr (PS bmr) under irrigated and dryland conditions during both years of this study. However, PS bmr biomass had higher cellulosic EtOH conversion efficiency than SP 1990 in both years according to simultaneous saccharification and fermentation analysis. Irrigation resulted in 26–49 % more biomass and 28–72 % more cellulosic EtOH production during both growing seasons, indicating that limited irrigation had favorable effects on both biomass and biofuel production. In the first year, when precipitation was below average, both cultivars produced similar amounts of cellulosic EtOH. During the second year, when precipitation was above average, higher biomass production of SP 1990 resulted in 28 % higher cellulosic EtOH production than PS bmr when averaged across both irrigated and dryland conditions. The large range of cellulosic EtOH production (1,600 to 3,380 L?ha^?1) during the 2 years of this study was primarily driven by differences in water availability that resulted from precipitation and irrigation. Our findings indicates that chemical composition and biomass yield potential of sorghum cultivars are critical factors that affect biomass and biofuel production under limited water conditions.     

Life Cycle Performance of Cellulosic Ethanol and Corn Ethanol from a Retrofitted Dry Mill Corn Ethanol Plant

This study conducts a life cycle assessment of a simulated dry mill corn ethanol facility in California’s Central Valley retrofitted to also produce ethanol from corn stover, a cellulosic feedstock. The assessment examines three facility designs, all producing corn ethanol and wet distiller’s grains and solubles as a co-product: a baseline facility with no cellulosic retrofit, a facility retrofitted with a small capacity for stover feedstock, and a facility retrofitted for a large capacity of stover feedstock. Corn grain is supplied by rail from the Midwest, while stover is sourced from in-state farms and delivered by truck. Two stover feedstock supply scenarios are considered, testing harvest rates at 25 or 40 % of stover mass. Allocation is required to separate impacts attributable to co-products. Additional scenarios are explored to assess the effect of co-product allocation methods on life cycle assessment results for the two fuel products, corn ethanol and stover ethanol. The assessment tracks greenhouse gas (GHG) emissions, energy consumption, criteria air pollutants, and direct water consumption. The GHG intensity of corn ethanol produced from the three facility designs range between 61.3 and 68.9 g CO₂e/MJ, which includes 19.8 g CO₂e/MJ from indirect land use change for Midwestern corn grain. The GHG intensity of cellulosic ethanol varies from 44.1 to 109.2 g CO₂e/MJ, and 14.6 to 32.1 g CO₂e/MJ in the low and high stover capacity cases, respectively. Total energy input ranges between 0.60 and 0.71 MJ/MJ for corn ethanol and 0.13 to 2.29 MJ/MJ for stover ethanol. This variability is the result of the stover supply scenarios (a function of harvest rate) and co-product allocation decisions.     

Integrated Assessment of Large-Scale Biofuel Production

This paper proposes a different framework for discussing the possibility of replacing a significant fraction of fossil energy consumption of modern economies with biofuels. The proposed analysis is not based on the two classic feasibility checks—land availability and output/input energy ratio—debated in the majority of the literature in this field. Rather, the focus is on the desirability of an energy sector powered by biomass energy. Discussing of desirability requires introducing a multicriteria approach, that in turn requires a definition of a set of criteria of performance for such an energy sector. The concepts of societal metabolism and ecosystem metabolism are introduced to define five criteria of performance for an energy sector powered by alternative sources.

This paper does not tell the society whether or not biofuels should be used to replace fossil energy. Rather, it proposes a method of characterization of pros and cons for the option biofuel which can be used to make more informed choices. An analysis of three systems of production—corn-ethanol, sunflower-biodiesel, and SRWC-methanol—is provided to indicate the existence of systemic characteristics associated with an energy sector powered by biofuels. These characteristics are likely to persist even when different technical coefficients will be achieved. The conclusion is that, at the moment, it is not possible to replace the actual performance of an energy sector based on fossil energy with an energy sector running on biofuel. Biomass energy can and will have to play an important role in the sustainability of humankind, but this will require a better understanding of (i) the role that an energy sector plays within a given structure of societal metabolism; and (ii) the impact generated on ecosystem metabolism by societal metabolism, plus a lot of wisdom.     

The Benefits of Harvesting Wetland Invaders for Cellulosic Biofuel: An Ecosystem Services Perspective

The emerging interest in cellulosic biofuel production has led the call for alternative cropping systems that maximize production along with the accompanying regulating, supporting, and cultural ecosystem services. We evaluate the potential for biomass harvested from invaded wetlands to achieve these goals. The ecosystem service trade‐offs associated with a wetland invader harvest are evaluated followed by a case study estimating the energy production and nutrient removal of harvesting Phalaris arundinacea from invaded wetlands in Wisconsin, United States. Estimates for energy production from this single species harvest dwarf current renewable energy sources for the state of Wisconsin and offer the potential to recapture approximately 50–200% of the excess nitrogen and phosphorus annually applied as fertilizer. This restoration technique would not only generate income from biomass sales to subsidize the cost of restoration, but also has the potential to shift the system toward more desirable environmental conditions by removing nutrients annually, reducing downstream eutrophication, and enhancing the ability of more desirable vegetation to establish by removing the litter layer created by the invasive species.     

C_4 Plants as Biofuel Feedstocks: Optimising Biomass Production and Feedstock Quality from a Lignocellulosic Perspective

The main feedstocks for bioethanol are sugarcane (Saccharum officinarum) and maize (Zea mays),both of which are C4 grasses,highly efficient at converting solar energy into chemical energy,and both are food crops.As the systems for lignocellulosic bioethanol production become more efficient and cost effective,plant biomass from any source may be used as a feedstock for bioethanol production.Thus,a move away from using food plants to make fuel is possible,and sources of biomass such as wood from forestry and ...     

Switchgrass Biofuel Production on Reclaimed Surface Mines: II. Feedstock Quality and Theoretical Ethanol Production

Switchgrass (Panicum virgatum L.), a warm-season perennial grass, is an important bioenergy crop candidate because it produces high biomass yields on marginal lands and on reclaimed surface mined sites. In companion studies, dry matter (DM) yields for Cave-in-Rock, Shawnee, and Carthage cultivars varied from 4.2 to 13.0 Mg ha^?1averaged over 6 years at the reclaimed Hampshire site, and fertilization increased yields of Cave-in-Rock at Black Castle and Coal Mac sites from 0.3 to 2 Mg ha^?1 during the first 3 years. The objective of these experiments was to compare the impacts of cultivar and soil amendments on biomass quality and theoretical ethanol production of switchgrass grown on surface mines with differing soil characteristics. Biomass quality was determined for fiber, ash, lignin, digestibility, and carbohydrate contents via near-infrared reflectance spectroscopy, and carbohydrates were used to calculate theoretical ethanol yield (TEY; L Mg^?1) and multiplied by biomass yield to calculate theoretical ethanol production (TEP; L ha^?1). Cultivars at the Hampshire site did not differ in TEY and ranged from 426 to 457 L Mg^?1. Theoretical ethanol production from Cave-in-Rock at Hampshire was 7350 L ha^?1, which was higher than other cultivars because of its greater biomass production. This TEP was higher than in other studies which predicted 4000 to 5000 L ha^?1. At the Black Castle and Coal Mac sites, fertilizer applications slightly affected biomass quality of switchgrass and TEY, but provided greater TEP as a function of increased yield. Similar to other findings, total switchgrass biomass production has more impact than compositional differences on TEP, so maximizing biomass production is critical for maximizing potential biofuel production. With appropriate soil substrates, fertilization, planning, and management, large areas of reclaimed surface mines can be converted to switchgrass stands to produce high biomass quality and yields to support a bioethanol industry.     

Switchgrass Biofuel Production on Reclaimed Surface Mines: I. Soil Quality and Dry Matter Yield

Growing food crops for biofuel on productive agricultural lands may become less viable as requirements to feed a growing human population increase. This has increased interest in growing cellulosic biofuel feedstocks on marginal lands. Switchgrass (Panicum virgatum L.), a warm-season perennial, is a viable bioenergy crop candidate because it produces high yields on marginal lands under low fertility conditions. In other studies, switchgrass dry matter (DM) yields on marginal croplands varied from 5.0 to 10.0 Mg ha^?1 annually. West Virginia contains immense areas of reclaimed surface mined lands that could support a switchgrass-based biofuel industry, but yield data on these lands are lacking. Field experiments were established in 2008 to determine yields of three switchgrass cultivars on two West Virginia mine sites. One site reclaimed with topsoil and municipal sludge produced biomass yields of 19.0 Mg DM ha^?1 for Cave-in-Rock switchgrass after the sixth year, almost double the varieties Shawnee and Carthage, at 10.0 and 5.7 Mg ha^?1, respectively. Switchgrass yields on another site with no topsoil were 1.0 Mg ha^?1 after the sixth year, with little variation among cultivars. A second experiment was conducted at two other mine sites with a layer of topsoil over gray overburden. Cave-in-Rock was seeded with fertilizer applications of 0, 34, and 68 kg N-P₂O₅-K₂O ha^?1. After the third year, the no fertilizer treatment averaged biomass yields of 0.3 Mg ha^?1, while responses to the other two rates averaged 1.1 and 2.0 Mg ha^?1, respectively. Fertilization significantly increased yields on reclaimed mine soils. Where mine soil fertility was good, yields were similar to those reported on agricultural soils in the Northeastern USA.     

Crop Management Impacts Biofuel Quality: Influence of Switchgrass Harvest Time on Yield, Nitrogen and Ash of Fast Pyrolysis Products

Although upgrading bio-oil from fast pyrolysis of biomass is an attractive pathway for biofuel production, nitrogen (N) and mineral matter carried over from the feedstock to the bio-oil represents a serious contaminant in the process. Reducing the N and ash content of biomass feedstocks would improve process reliability and reduce production costs of pyrolytic biofuels. This study investigated: (1) How does switchgrass harvest date influence the yield, N concentration ([N]), and ash concentration of biomass and fast pyrolysis products? and (2) Is there a predictive relationship between [N] of switchgrass biomass and [N] of fast pyrolysis products? Switchgrass from five harvest dates and varying [N] from central Iowa were pyrolyzed using a free-fall reactor. Harvestable biomass peaked in August (8.6 Mg ha^?1), dropping significantly by November (6.7 Mg ha^?1, P?=?0.0027). Production of bio-oil per unit area mirrored that of harvested biomass at each harvest date; however, bio-oil yield per unit dry biomass increased from 46.6 % to 56.7 % during the season (P?=?0.0018). Allowing switchgrass to senesce lowered biomass [N] dramatically, by as much as 68 % from June to November (P?<?0.0001). Concurrently, bio-oil [N] declined from 0.51 % in June to 0.17 % by November (P?<?0.0001). Significant reductions in ash concentration were also observed in biomass and char. Finally, we show for the first time that the [N] of switchgrass biomass is a strong predictor of the [N] of bio-oil, char, and non-condensable gas with R ² values of 0.89, 0.94, and 0.88, respectively.     

Modeled Impacts of Cover Crops and Vegetative Barriers on Corn Stover Availability and Soil Quality

Environmentally benign, economically viable, and socially acceptable agronomic strategies are needed to launch a sustainable lignocellulosic biofuel industry. Our objective was to demonstrate a landscape planning process that can ensure adequate supplies of corn (Zea mays L.) stover feedstock while protecting and improving soil quality. The Landscape Environmental Assessment Framework (LEAF) was used to develop land use strategies that were then scaled up for five U.S. Corn Belt states (Nebraska, Iowa, Illinois, Indiana, and Minnesota) to illustrate the impact that could be achieved. Our results show an annual sustainable stover supply of 194 million Mg without exceeding soil erosion T values or depleting soil organic carbon [i.e., soil conditioning index (SCI)?>?0] when no-till, winter cover crop, and vegetative barriers were incorporated into the landscape. A second, more rigorous conservation target was set to enhance soil quality while sustainably harvesting stover. By requiring erosion to be <1/2 T and the SCI-organic matter (OM) subfactor to be >?0, the annual sustainable quantity of harvestable stover dropped to148 million Mg. Examining removal rates by state and soil resource showed that soil capability class and slope generally determined the effectiveness of the three conservation practices and the resulting sustainable harvest rate. This emphasizes that sustainable biomass harvest must be based on subfield management decisions to ensure soil resources are conserved or enhanced, while providing sufficient biomass feedstock to support the economic growth of bioenergy enterprises.     

Cellulosic Butanol (ABE) Biofuel Production from Sweet Sorghum Bagasse (SSB): Impact of Hot Water Pretreatment and Solid Loadings on Fermentation Employing <Emphasis Type="Italic">Clostridium beijerinckii</Emphasis> P260

A novel butanol fermentation process was developed in which sweet sorghum bagasse (SSB) was pretreated using liquid hot water (LHW) pretreatment technique followed by enzymatic hydrolysis and butanol (acetone butanol ethanol (ABE)) fermentation. A pretreatment temperature of 200 °C resulted in the generation of a hydrolyzate that inhibited butanol fermentation. When SSB pretreatment temperature was decreased to 190 °C (0-min holding time), the hydrolyzate was successfully fermented without inhibition and an ABE productivity of 0.51 g L^?1 h^?1 was achieved which is comparable to the 0.49 g L^?1 h^?1 observed in the control fermentation where glucose was used as a feedstock. These results are based on the use of 86 g L^?1 SSB solid loadings in the pretreatment reactors. We were also able to increase SSB solid loadings from 120 to 200 g L^?1 in the pretreatment step (190 °C) followed by hydrolysis and butanol fermentation. As pretreatment solid loadings increased, ABE yield remained in the range of 0.38–0.46. In these studies, a maximum ABE concentration of 16.88 g L^?1 was achieved. Using the LHW pretreatment technique, 88.40–96.00 % of polymeric sugars (cellulose + hemicellulose) were released in the SSB hydrolyzate. The LHW pretreatment technique does not require chemical additions and is environmentally friendly, and the hydrolyzate can be used successfully for butanol fermentation.     

Direct and Efficient Production of Ethanol from Cellulosic Material with a Yeast Strain Displaying Cellulolytic Enzymes

For direct and efficient ethanol production from cellulosic materials, we constructed a novel cellulose-degrading yeast strain by genetically codisplaying two cellulolytic enzymes on the cell surface of Saccharomyces cerevisiae. By using a cell surface engineering system based on α-agglutinin, endoglucanase II (EGII) from the filamentous fungus Trichoderma reesei QM9414 was displayed on the cell surface as a fusion protein containing an RGSHis6 (Arg-Gly-Ser-His₆) peptide tag in the N-terminal region. EGII activity was detected in the cell pellet fraction but not in the culture supernatant. Localization of the RGSHis6-EGII-α-agglutinin fusion protein on the cell surface was confirmed by immunofluorescence microscopy. The yeast strain displaying EGII showed significantly elevated hydrolytic activity toward barley β-glucan, a linear polysaccharide composed of an average of 1,200 glucose residues. In a further step, EGII and β-glucosidase 1 from Aspergillus aculeatus No. F-50 were codisplayed on the cell surface. The resulting yeast cells could grow in synthetic medium containing β-glucan as the sole carbon source and could directly ferment 45 g of β-glucan per liter to produce 16.5 g of ethanol per liter within about 50 h. The yield in terms of grams of ethanol produced per gram of carbohydrate utilized was 0.48 g/g, which corresponds to 93.3% of the theoretical yield. This result indicates that efficient simultaneous saccharification and fermentation of cellulose to ethanol are carried out by a recombinant yeast cells displaying cellulolytic enzymes.     

宏基因组技术在新型生物燃料生产酶制剂开发中的应用

随着石化燃料的日益减少,以植物生物质为原料的可再生生物燃料成为石化燃料的理想替代品。然而微生物降解生物质效率低下,是生物燃料生产过程中一大难题,因此开发效率高、稳定性强的微生物酶制剂显得尤为重要。近年来,宏基因组技术的发展为生物燃料的生产提供了多种新型酶制剂。宏基因组技术是直接提取环境样品中的总DNA,通过构建文库,筛选目的基因或功能基因的方法,在用于燃料生产的新型酶制剂的开发中发挥着重要作用。本文概述了宏基因组技术的实施策略,总结了包括纤维素酶、蛋白酶、酯酶、脂肪酶等多种酶资源开发的最新研究进展,并综合和讨论了通过酶法将木质纤维素等生物材料有效转化为生物燃料的途径,为新酶的开发提供了新思路。