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The isomerization of xylose to xylulose is considered the most promising approach to initiate xylose bioconversion. Here, phylogeny-guided big data mining, rational modification, and ancestral sequence reconstruction strategies were implemented to explore new active xylose isomerases (XIs) for Saccharomyces cerevisiae . Significantly, 13 new active XIs for S. cerevisiae were mined or artificially created. Moreover, the importance of the amino-terminal fragment for maintaining basic XI activity was demonstrated. With the mined XIs, four efficient xylose-utilizing S. cerevisiae were constructed and evolved, among which the strain S. cerevisiae CRD5HS contributed to ethanol titers as high as 85.95 and 94.76 g/liter from pretreated corn stover and corn cob, respectively, without detoxifying or washing pretreated biomass. Potential genetic targets obtained from adaptive laboratory evolution were further analyzed by sequencing the high-performance strains. The combined XI mining methods described here provide practical references for mining other scarce and valuable enzymes.

A novel ammonia-based pretreatment for densified lignocellulosic biomass was developed to reduce ammonia usage and integrate with viable biomass logistics scenarios. The COmpacted Biomass with Recycled Ammonia (COBRA) pretreatment performed at 100 degrees C allows >95% conversion of sugarcane bagasse (SCB) carbohydrates into soluble monomeric and oligomeric sugars (glucose and xylose) using industrially relevant 6% glucan loading (similar to 21% solids loading) enzymatic hydrolysis conditions at reduced enzyme loadings. Pretreatment via COBRA with simultaneous lignin extraction (COBRA-LE) improved Saccharomyces cerevisiae 424A(LNH-ST) metabolic yield from 89% to 97.5% relative to COBRA without delignification, allowing a process ethanol yield of 71.6%. A technoeconomic analysis on SCB biorefining to ethanol in the state of Sao Paulo, Brazil, compared COBRA to other mature technologies, such as AFEX and steam-explosion. Amongst all scenarios studied, biorefineries based on COBRA-LE pretreatment offered the lowest average minimum ethanol selling price of US$1.45 per gallon ethanol. COBRA pretreatment was subsequently tested on perennial grasses and hardwoods, and >80% total sugar yields were achieved for all cases. info:eu-repo/semantics/publishedVersion

Achievement of the 1.5 °C limit for global temperature increase relies on the large-scale deployment of carbon dioxide removal (CDR) technologies. In this article, we explore two CDR technologies: soil carbon sequestration (SCS), and carbon capture and storage (CCS) integrated with cellulosic biofuel production. These CDR technologies are applied as part of decentralized biorefinery systems processing corn stover and unfertilized switchgrass grown in riparian zones in the Midwestern United States. Cover crops grown on corn-producing lands are chosen from the SCS approach, and biogenic CO2 in biorefineries is captured, transported by pipeline, and injected into saline aquifers. The decentralized biorefinery system using SCS, CCS, or both can produce carbon-negative cellulosic biofuels (≤-22.2 gCO2 MJ-1). Meanwhile, biofuel selling prices increase by 15-45% due to CDR costs. Economic incentives (e.g., cover crop incentives and/or a CO2 tax credit) can mitigate price increases caused by CDR technologies. A combination of different CDR technologies in decentralized biorefinery systems is the most efficient method for greenhouse gas (GHG) mitigation, and its total GHG mitigation potential in the Midwest is 0.16 GtCO2 year-1.

Plant biomasses enriched in crystalline cellulose allomorphs, such as native cellulose I (CI), can be structurally altered using anhydrous liquid ammonia to form an enzymatically less recalcitrant ...

The current or “conventional” paradigm for producing process energy in a biorefinery processing cellulosic biomass is on‐site energy recovery through combustion of residual solids and biogas generated by the process. Excess electricity is then exported, resulting in large greenhouse gas (GHG) credits. However, this approach will cause lifecycle GHG emissions of biofuels to increase as more renewable energy sources (wind, solar, etc.) participate in grid‐electricity generation, and the GHG credits from displacing fossil fuel decrease. To overcome this drawback, a decentralized (depot‐based) biorefinery can be integrated with a coal‐fired power plant near a large urban area. In an integrated, decentralized, depot‐based biorefinery (IDB), the residual solids are co‐fired with coal either in the adjacent power plant or in coal‐fired boilers elsewhere to displace coal. An IDB system does not rely on indirect GHG credits through grid‐electricity displacement. In an IDB system, biogas from the wastewater treatment facility is also upgraded to biomethane and used as a transportation biofuel. The GHG savings per unit of cropland in the IDB systems (2.7–2.9 MgCO2/ha) are 1.5–1.6 fold greater than those in a conventional centralized system (1.7–1.8 MgCO2/ha). Importantly, the biofuel selling price in the IDBs is lower by 28–30 cents per gasoline‐equivalent liter than in the conventional centralized system. Furthermore, the total capital investment per annual biofuel volume in the IDB is much lower (by ~80%) than that in the conventional centralized system. Therefore, utilization of biomethane and residual solids in the IDB systems leads to much lower biofuel selling prices and significantly greater GHG savings per unit of cropland participating in the biorefinery system compared to the conventional centralized biorefineries.

Biological lignin valorization has emerged as a major solution for sustainable and cost-effective biorefineries. However, current biorefineries yield lignin with inadequate fractionation for bioconversion, yet substantial changes of these biorefinery designs to focus on lignin could jeopardize carbohydrate efficiency and increase capital costs. We resolve the dilemma by designing ‘plug-in processes of lignin’ with the integration of leading pretreatment technologies. Substantial improvement of lignin bioconversion and synergistic enhancement of carbohydrate processing are achieved by solubilizing lignin via lowering molecular weight and increasing hydrophilic groups, addressing the dilemma of lignin- or carbohydrate-first scenarios. The plug-in processes of lignin could enable minimum polyhydroxyalkanoate selling price at as low as $6.18/kg. The results highlight the potential to achieve commercial production of polyhydroxyalkanoates as a co-product of cellulosic ethanol. Here, we show that the plug-in processes of lignin could transform biorefinery design toward sustainability by promoting carbon efficiency and optimizing the total capital cost., The current biorefineries yield lignin with inadequate fractionation for bioconversion, yet substantial changes of these biorefinery designs could jeopardize carbohydrate efficiency and increase capital costs. Here the authors resolve the dilemma by designing ‘plug-in processes of lignin’ to enable economic waste valorization.

Current food production practices tend to damage and deplete soil, diminish biodiversity, and degrade water supplies. For agriculture to become environmentally sustainable and simultaneously increase food output for a growing world population, fundamental changes in agricultural production systems are required. Renewable energy can reduce greenhouse gases (GHGs) but we also need simple, low‐cost approaches to remove atmospheric carbon and sequester it in stable forms. Recycling of digestate from the anaerobic digestion of agricultural and waste materials to soils can sequester atmospheric carbon and provide many other economic, social and environmental benefits. Biogasdoneright™ (BDR) is a set of practices that link biogas production with sustainable agriculture. The BDR approach to sustainable agriculture is being implemented on a large scale in Italy. In this paper, we examine the potential impact of implementing BDR in selected other countries. The biomethane potential in these countries, estimated conservatively, varies from about 10–30% of their current annual natural gas consumption. Biomethane from sequential (double) crops provides by far the greatest fraction of the biomethane potential. Double cropping also drives many of the environmental and economic benefits of BDR systems. Depending on where and how widely it is implemented, the production of biogas in BDR systems could have very significant national‐level impacts. For example, sufficient biomethane could be produced in Argentina to completely eliminate imports of natural gas, equivalent to about 28% of Argentina's 2017 trade deficit. In the USA, renewable biogas could generate electricity equal to nearly all of the electricity currently produced by domestic solar and wind resources. © 2020 Society of Chemical Industry and John Wiley & Sons, Ltd

New models for renewable energy production are needed to simultaneously decrease greenhouse gases (GHG) emissions, use land more efficiently and replace large amounts of fossil fuel. Ethanol production and livestock feed integration as practiced in the United States (USA) is one model for ethanol production combined with animal feed production. Brazil, the second largest ethanol and beef cattle producer in the world, can adapt the USA model of corn ethanol and cattle integration considering its local characteristics. This paper evaluates the techno-economic and environmental feasibility of sugarcane ethanol and cattle integration, thereby avoiding pasture displacement into forests or other sensitive lands. Cattle can be fattened in feedlots using some sugarcane ethanol byproducts. Intensification of cattle production by integration with sugarcane production releases pasture area to produce more biofuels, without needing more land for cattle production. The release of pasture land to produce more sugarcane results in what we call “avoided ILUC”, the resultant reduced GHG emissions compared to conventional sugarcane ethanol, because no additional land is needed to accommodate an additional sugarcane ethanol production. Simulations were performed using the Virtual Sugarcane Biorefinery (VSB) model developed by the Brazilian Bioethanol Science and Technology Laboratory (CTBE). We calculated as economic parameters the internal rate of return (IRR), net present value (NPV) and payback time. Climate impacts were assessed via Life Cycle Assessment. Sugarcane and cattle integration decreases overall climate impacts compared to non-integrated systems. Techno-economic feasibility is achieved by additional land rental revenues for released pasture area and by carbon credits.

Lignocellulosic materials are plant-derived feedstocks, such as crop residues (e.g., corn stover, rice straw, and sugar cane bagasse) and purpose-grown energy crops (e.g., miscanthus, and switchgrass) that are available in large quantities to produce biofuels, biochemicals, and animal feed. Plant polysaccharides (i.e., cellulose, hemicellulose, and pectin) embedded within cell walls are highly recalcitrant towards conversion into useful products. Ammonia fiber expansion (AFEX) is a thermochemical pretreatment that increases accessibility of polysaccharides to enzymes for hydrolysis into fermentable sugars. These released sugars can be converted into fuels and chemicals in a biorefinery. Here, we describe a laboratory-scale batch AFEX process to produce pretreated biomass on the gram-scale without any ammonia recycling. The laboratory-scale process can be used to identify optimal pretreatment conditions (e.g., ammonia loading, water loading, biomass loading, temperature, pressure, residence time, etc.) and generates sufficient quantities of pretreated samples for detailed physicochemical characterization and enzymatic/microbial analysis. The yield of fermentable sugars from enzymatic hydrolysis of corn stover pretreated using the laboratory-scale AFEX process is comparable to pilot-scale AFEX process under similar pretreatment conditions. This paper is intended to provide a detailed standard operating procedure for the safe and consistent operation of laboratory-scale reactors for performing AFEX pretreatment of lignocellulosic biomass.

Alkali and acid pretreatment based processes for lignocellulosic biofuel production typically consume substantial amount of alkali and acid. In addition, alkali pretreatment generates compounds with high toxicity rendering washing/detoxification inevitable. In this work, alkali (AL) and dilute acid (DA) pretreatment based processes were compared side by side for ethanol production from corn stover (CS) using commercial enzymes and Zymomonas mobilis 8b strain. Next, AL and DA pretreated CS slurries (without removing liquid stream) were mixed to neutralize for enzymatic hydrolysis and ethanol fermentation. The digestibility and fermentability of the mixed pretreated biomass were systematically studied and compared to unmixed pretreated biomass. Results showed that mixed biomass process saved 40–50% alkali/acid use and enhanced fermentability compared to AL-CS. This work suggests that it could be beneficial to have both alkali and acid pretreatments operating in a biorefinery, which could accommodate various feedstocks.

Diverse lignin samples have been subjected to a catalytic aerobic oxidation process, followed by formic-acid-induced hydrolytic depolymerization. The yield of monomeric aromatic compounds varies depending on the lignin plant source and pretreatment method. The best results are obtained from poplar lignin isolated via a acidolysis pretreatment method, which gives 42 wt% yield of low-molecular-weight aromatics. Use of other pretreatment methods and/or use of maple and maize lignins afford yields of aromatics ranging from 3 to 31 wt%. These results establish useful references for the development of improved oxidation/depolymerization protocols.

It has been previously shown that cellulose-lignin droplets’ strong interactions, resulting from lignin coalescence and redisposition on cellulose surface during thermochemical pretreatments, increase cellulose recalcitrance to biological conversion, especially at commercially viable low enzyme loadings. However, information on the impact of cellulose–hemicellulose interactions on cellulose recalcitrance following relevant pretreatment conditions are scarce. Here, to investigate the effects of plausible hemicellulose precipitation and re-association with cellulose on cellulose conversion, different pretreatments were applied to pure Avicel® PH101 cellulose alone and Avicel mixed with model hemicellulose compounds followed by enzymatic hydrolysis of resulting solids at both low and high enzyme loadings. Solids produced by pretreatment of Avicel mixed with hemicelluloses (AMH) were found to contain about 2 to 14.6% of exogenous, precipitated hemicelluloses and showed a remarkably much lower digestibility (up to 60%) than their respective controls. However, the exogenous hemicellulosic residues that associated with Avicel following high temperature pretreatments resulted in greater losses in cellulose conversion than those formed at low temperatures, suggesting that temperature plays a strong role in the strength of cellulose–hemicellulose association. Molecular dynamics simulations of hemicellulosic xylan and cellulose were found to further support this temperature effect as the xylan–cellulose interactions were found to substantially increase at elevated temperatures. Furthermore, exogenous, precipitated hemicelluloses in pretreated AMH solids resulted in a larger drop in cellulose conversion than the delignified lignocellulosic biomass containing comparably much higher natural hemicellulose amounts. Increased cellulase loadings or supplementation of cellulase with xylanases enhanced cellulose conversion for most pretreated AMH solids; however, this approach was less effective for solids containing mannan polysaccharides, suggesting stronger association of cellulose with (hetero) mannans or lack of enzymes in the mixture required to hydrolyze such polysaccharides.

Liquid fossil fuels (1) enable transportation and (2) provide energy for mobile work platforms and (3) supply dispatchable energy to highly variable demand (seasonal heating and peak electricity). We describe a system to replace liquid fossil fuels with drop-in biofuels including gasoline, diesel and jet fuel. Because growing biomass removes carbon dioxide from the air, there is no net addition of carbon dioxide to the atmosphere from burning biofuels. In addition, with proper management, biofuel systems can sequester large quantities of carbon as soil organic matter, improving soil fertility and providing other environmental services. In the United States liquid biofuels can potentially replace all liquid fossil fuels. The required system has two key features. First, the heat and hydrogen for conversion of biomass into high-quality liquid fuels is provided by external low-carbon energy sources--nuclear energy or fossil fuels with carbon capture and sequestration. Using external energy inputs can almost double the energy content of the liquid fuel per unit of biomass feedstock by fully converting the carbon in biomass into a hydrocarbon fuel. Second, competing effectively with fossil fuels requires very large biorefineries—the equivalent of a 250,000 barrel per day oil refinery. This requires commercializing methods for converting local biomass into high-density storable feedstocks that can be economically shipped to large-scale biorefineries.

Bioenergy has been under intense scrutiny over the last ten years with significant research efforts in many countries taking place to define and measure sustainable practices. We describe here the main challenges and policy issues and provide policy recommendations for scaling up sustainable bioenergy approaches globally. The 2016 Intended Nationally Determined Contributions (INDCs defined under the UN Framework Convention on Climate Change) (UNFCCC) Conference of the Parties (COP21) will not reach global Greenhouse Gas (GHG) emission targets of 2 °C. Sustainable biomass production can make a significant contribution. Substantive evidence exists that many bioenergy cropping systems can bring multiple benefits and off-set environmental problems associated with fossil fuels usage as well as intensive food production and urbanization. We provide evidence that there are many approaches to land use for bioenergy expansion that do not lead to competition for food or other needs. We should focus on how to manage these approaches on a synergistic basis and how to reduce tradeoffs at landscape scales. Priorities include successful synergies between bioenergy and food security (integrated resource management designed to improve both food security and access to bioenergy), investments in technology, rural extension, and innovations that build capacity and infrastructure, promotion of stable prices to incentivize local production and use of double cropping and flex crops (plants grown for both food and non-food markets) that provide food and energy as well as other services. The sustainable production of biomass requires appropriate policies to secure long-term support to improve crop productivity and also to ensure environmental as well as economic and social benefits of bioenergy cropping systems. Continuous support for cropping, infrastructure, agricultural management and related policies is needed to foster positive synergies between food crops and bioenergy production. In comparison to fossil fuels, biofuels have many positive environmental benefits. Potential negative effects caused by land-use change and agriculture intensification can be mitigated by agroecological zoning, best management practices, the use of eco-hydrology and biodiversity-friendly concepts at field, watershed and landscape scales. Global climate and environmental changes related to the use of fossil fuels and inequitable development make it unethical not to pursue more equitable energy development that includes bioenergy. To achieve sustainable development, competitiveness and costs of bioenergy production need to be addressed in a manner that considers not only economic gains but also development of local knowledge and social and environmental benefits.

We reported previously on the Biogasdoneright™ system for on-farm biogas production. This innovative system employs sequential (year-round) cropping to produce both food and energy from agricultural biomass, primarily cellulosic materials. This paper uses a marginal analysis approach to estimate the life cycle greenhouse gas emissions (GHGs) of electricity and biomethane produced by four currently operating Italian biogas plants that process various agricultural feedstocks, residues, and by-products. The biogas is burned on-farm to generate electricity that is then exported to the grid. The marginal lifecycle GHGs of this farm-produced electricity range from -335 to 25 grams CO2 per kilowatt hour (kWh). By comparison, the marginal GHGs of electricity generated by fossil fuels in the European Union (EU) is 752 grams CO2 per kWh. The biogas might also be upgraded to produce pipeline-quality biomethane, a direct substitute for natural gas. The marginal lifecycle GHGs of biomethane potentially produced from the Biogasdoneright™ plants range from 10 to -36 grams CO2 per megajoule (MJ) while the corresponding figure for a conventional biogas plant is 27 grams CO2 per MJ. Natural gas in the EU produces 72 grams CO2 per MJ and marginal fossil fuel in the EU generates 115 grams CO2 per MJ. Negative GHG emissions arise largely from avoided emissions of agricultural effluents and residues. © 2017 Society of Chemical Industry and John Wiley & Sons, Ltd

High solids loadings (>18 wt%) in enzymatic hydrolysis and fermentation are desired for lignocellulosic biofuel production at a high titer and low cost. However, sugar conversion and ethanol yield decrease with increasing solids loading. The factor(s) limiting sugar conversion at high solids loading is not clearly understood. In the present study, we investigated the effect of solids loading on simultaneous saccharification and co-fermentation (SSCF) of AFEX™ (ammonia fiber expansion) pretreated corn stover for ethanol production using a xylose fermenting strain Saccharomyces cerevisiae 424A(LNH-ST). Decreased sugar conversion and ethanol yield with increasing solids loading were also observed. End-product (ethanol) was proven to be the major cause of this issue and increased degradation products with increasing solids loading was also a cause. For the first time, we show that with in situ removal of end-product by performing SSCF aerobically, sugar conversion stopped decreasing with increasing solids loading and monomeric sugar conversion reached as high as 93% at a high solids loading of 24.9 wt%. Techno-economic analysis was employed to explore the economic possibilities of cellulosic ethanol production at high solids loadings. The results suggest that low-cost in situ removal of ethanol during SSCF would significantly improve the economics of high solids loading processes. Biotechnol. Bioeng. 2017;114: 980-989. © 2016 Wiley Periodicals, Inc.

Bacterial protein secretion represents a significant challenge in biotechnology, which is essential for the cost‐effective production of therapeutics, enzymes, and other functional proteins. Here, it is demonstrated that proteomics‐guided engineering of transcription, translation, secretion, and folding of ligninolytic laccase balances the process, minimizes the toxicity, and enables efficient heterologous secretion with a total protein yield of 13.7 g L−1. The secretory laccase complements the biochemical limits on lignin depolymerization well in Rhodococcus opacus PD630. Further proteomics analysis reveals the mechanisms for the oleaginous phenotype of R. opacus PD630, where a distinct multiunit fatty acid synthase I drives the carbon partition to storage lipid. The discovery guides the design of efficient lipid conversion from lignin and carbohydrate. The proteomics‐guided integration of laccase‐secretion and lipid production modules enables a high titer in converting lignin‐enriched biorefinery waste to lipid. The fundamental mechanisms, engineering components, and design principle can empower transformative platforms for biomanufacturing and biorefining.

The Renewable Fuel Standard (RFS) program specifies a greenhouse gas (GHG) reduction threshold for cellulosic biofuels, while the Low Carbon Fuel Standard (LCFS) program in California does not. Here, we investigate the effects of the GHG threshold under the RFS on projected GHG savings from two corn stover-based biofuel supply chain systems in the United States Midwest. The analysis is based on a techno-economic framework that minimizes ethanol selling price. The GHG threshold lowers the lifecycle GHG of ethanol: 34.39 ± 4.92 gCO

A sustainable chemical industry cannot exist at scale without both sustainable feedstocks and feedstock supply chains to provide the raw materials. However, most current research focus is on producing the sustainable chemicals and materials. Little attention is given to how and by whom sustainable feedstocks will be supplied. In effect, we have put the bioproducts cart before the sustainable feedstocks horse. For example, bulky, unstable, non-commodity feedstocks such as crop residues probably cannot supply a large-scale sustainable industry. Likewise, those who manage land to produce feedstocks must benefit significantly from feedstock production, otherwise they will not participate in this industry and it will never grow. However, given real markets that properly reward farmers, demand for sustainable bioproducts and bioenergy can drive the adoption of more sustainable agricultural and forestry practices, providing many societal “win–win” opportunities. Three case studies are presented to show how this “win–win” process might unfold.

Corn stover supply chains in a distributed biorefinery system are explored. The distributed cellulosic biorefinery uses pre-processed and densified cellulosic feedstock from a geographically separated facility (a depot) as raw material. A network of small-scale depot facilities supplies pre-processed feedstock to a distributed biorefinery. Depot facilities are assumed to be located at existing grain elevators, while distributed biorefineries are located adjacent to coal-fired power plants in areas with high gasoline consumption (urban areas) in the Midwest. The county level corn stover projections in 2022 by the US Billion-Ton Update report (2011) are used to estimate ethanol selling price and greenhouse gas (GHG) emissions of the ethanol fuel. The supply chain for each distributed biorefinery is determined by minimizing the ethanol selling price. Approximately ten distributed biorefineries based on corn stover could be established in the Midwest. Over 700 individual depot facilities participate in supplying the distributed biorefinery systems which collectively can produce greater than 12 hm3 of ethanol (3.3 billion gallons) per year. Ethanol selling price in the distributed system ranges from US$0.66 to US$1.03 per liter. Some distributed biorefineries are economically competitive with a centralized biorefinery. However, not every region can support a distributed biorefinery system due to inadequate corn stover availability. Cradle-to-gate GHG emissions of ethanol in the distributed systems are 22.1–46.6 g CO2 per MJ. The external energy consumption in the depot facilities is the major GHG source. Optimizing process energy use in the depot facility is required to reduce both operation costs and GHG emissions. © 2016 Society of Chemical Industry and John Wiley & Sons, Ltd

Oil palm empty fruit bunch (OPEFB) fibre is widely available in Southeast Asian countries and found to have 60% (w/w) sugar components. OPEFB was pretreated using the ammonia fibre expansion (AFEX) method and characterised physically by the Fourier transform infrared spectroscopy, X-ray diffraction and scanning electron microscopy. The results show that there were significant structural changes in OPEFB after the pretreatment step, and the sugar yield after enzymatic hydrolysis using a cocktail of Cellic Ctec2® and Cellic Htec2® increased from 0.15gg(-1) of OPEFB in the raw untreated OPEFB sample to 0.53gg(-1) of OPEFB in AFEX-pretreated OPEFB (i.e. almost a fourfold increase in sugar conversion), which enhances the economic value of OPEFB. A biohydrogen fermentability test of this hydrolysate was carried out using a locally isolated bacterium, Enterobacter sp. KBH6958. The biohydrogen yield after 72h of fermentation was 1.68mol H2 per mol sugar. Butyrate, ethanol, and acetate were the major metabolites.

A comparative analysis of the response of agave bagasse (AGB) to pretreatment by ammonia fiber expansion (AFEX™), autohydrolysis (AH) and ionic liquid (IL) was performed using 2D nuclear magnetic resonance (NMR) spectroscopy, wet chemistry, enzymatic saccharification and mass balances. It has been found that AFEX pretreatment preserved all carbohydrates in the biomass, whereas AH removed 62.4% of xylan and IL extracted 25% of lignin into wash streams. Syringyl and guaiacyl lignin ratio of untreated AGB was 4.3, whereas for the pretreated biomass the ratios were 4.2, 5.0 and 4.7 for AFEX, AH and IL, respectively. Using NMR spectra, the intensity of β-aryl ether units in aliphatic, anomeric, and aromatic regions decreased in all three pretreated samples when compared to untreated biomass. Yields of glucose plus xylose in the major hydrolysate stream were 42.5, 39.7 and 26.9kg per 100kg of untreated AGB for AFEX, IL and AH, respectively.

A group of over 600 Italian farmers organized as the Italian Biogas Consortium are redesigning their own farming systems to produce food and bioenergy in a nationwide farm-level movement called Biogasdoneright™. This Feature demonstrates how it is possible to simultaneously increase the economic viability and stability of agriculture by reducing farm input costs and enabling farmers to produce food and fuel more sustainably. © 2016 Society of Chemical Industry and John Wiley & Sons, Ltd

Universal access to modern energy is required if the world is to reduce poverty and enable sustainable development. Energy poverty is the lack of access to safe and efficient energy carriers. It affects more than 1.3 billion people without electricity and 2.7 billion without appropriate cooking facilities and fuels, particularly in sub-Saharan Africa (SSA). Under these conditions, people rely on traditional biomass to fulfil basic needs. Traditional biomass use causes death and disease associated with indoor air pollution, high labor demands to obtain biomass, and environmental damage due to deforestation. This paper analyzes the potential for modern energy production based on sugarcane in SSA. A standard, business-as-usual (BAU), sugar-based project is explored vis-a-vis more aggressive scenarios for producing modern energy, including electricity, ethanol, and solid fuel pellets. All scenarios considered are based on a single sugarcane mill processing one million tonnes of cane per year, grown in an area of 15 000 hectares. Our simulations show that over 210 000 households could be served with electricity and other 31 000 with modern cooking fuels under the scenarios examined. Less dependence on traditional biomass may also spare wooded environments from deforestation. However, harnessing modern energy from sugarcane does not come without challenges. Economic pitfalls (e.g. investment costs and affordability) coupled with poor political environments are among the main obstacles. Nevertheless, encouraging local and regional trends proved bioenergy a feasible way out of energy poverty and an alternative to sustainable development. © 2016 Society of Chemical Industry and John Wiley & Sons, Ltd

Oleaginous yeasts can convert sugars to lipids with fatty acid profiles similar to those of vegetable oils, making them attractive for production of biodiesel. Lignocellulosic biomass is an attractive source of sugars for yeast lipid production because it is abundant, potentially low cost, and renewable. However, lignocellulosic hydrolyzates are laden with byproducts which inhibit microbial growth and metabolism. With the goal of identifying oleaginous yeast strains able to convert plant biomass to lipids, we screened 32 strains from the ARS Culture Collection, Peoria, IL to identify four robust strains able to produce high lipid concentrations from both acid and base-pretreated biomass. The screening was arranged in two tiers using undetoxified enzyme hydrolyzates of ammonia fiber expansion (AFEX)-pretreated cornstover as the primary screening medium and acid-pretreated switch grass as the secondary screening medium applied to strains passing the primary screen. Hydrolyzates were prepared at ∼18–20% solids loading to provide ∼110 g/L sugars at ∼56:39:5 mass ratio glucose:xylose:arabinose. A two stage process boosting the molar C:N ratio from 60 to well above 400 in undetoxified switchgrass hydrolyzate was optimized with respect to nitrogen source, C:N, and carbon loading. Using this process three strains were able to consume acetic acid and nearly all available sugars to accumulate 50–65% of cell biomass as lipid (w/w), to produce 25–30 g/L lipid at 0.12–0.22 g/L/h and 0.13–0.15 g/g or 39–45% of the theoretical yield at pH 6 and 7, a performance unprecedented in lignocellulosic hydrolyzates. Three of the top strains have not previously been reported for the bioconversion of lignocellulose to lipids. The successful identification and development of top-performing lipid-producing yeast in lignocellulose hydrolyzates is expected to advance the economic feasibility of high quality biodiesel and jet fuels from renewable biomass, expanding the market potential for lignocellulose-derived fuels beyond ethanol for automobiles to the entire U.S. transportation market. Biotechnol. Bioeng. 2016;113: 1676–1690. © 2016 Wiley Periodicals, Inc.

Literature values for glucose release from corn stover are highly variable which would likely result in tremendous variability in bio-refinery ethanol yield from corn stover feedstock. A relatively recent change in United States corn genetics is the inclusion of the Bacillus thuringiensis (Bt) trait, which now accounts for three-fourths of all US planted corn acreage. The objective of this study was to evaluate the effect of corn grain yield, inclusion of the Bt trait, and location environment on corn stover quality for subsequent ethanol conversion. Two hybrid pairs (each having a Bt and non-Bt near-isoline) were analyzed giving a total of 4 hybrids. In 2010 and 2011, field plots were located in Michigan at four latitudinal differing locations in four replicated plots at each location. Stover composition and enzymatic digestibility was analyzed and estimated ethanol yield (g g −1 ) was calculated based on hydrolyzable glucan and xylan levels. Analysis showed that there were no significant differences in total glucose or xylose levels nor in enzymatically hydrolyzable glucan and xylan concentrations between Bt corn stover and the non-Bt stover isolines. Regression analyses between corn grain yield (Mg ha −1 ) and corn stover ethanol yield (g g −1 ) showed an inverse relationship indicative of a photosynthate source-sink relationship. Nevertheless, the quantity of stover produced was found to be more critical than the quality of stover produced in maximizing potential stover ethanol yield on a land area basis.

This study assesses the role of spatial-resolution and spatial-variations in environmental impacts estimation and decision-making for corn-stover harvesting to produce biofuels. Geospatial corn-stover yields and environmental impacts [global warming potential (GWP), eutrophication, and soil-loss] dataset for two study areas in Wisconsin and Michigan were generated through Environmental Policy Integrated Climate (EPIC) model and aggregated at different spatial-resolutions (i.e., 100; 1000; 10,000 ha). For each spatial-resolution, decision-making was accomplished using an optimization routine to minimize different environmental impacts associated with harvesting stover to meet varied biomass demands. The results of the study showed that selective harvesting at higher-resolution (or lower-aggregation level) can result in significantly lower environmental impacts, especially at low stover demand levels. Additionally, the increased spatial resolution had more impact in minimizing the environmental impacts of corn stover harvest under a more variable landscape such as terrains and its influences are more pronounced for soil-loss and eutrophication potential compared to GWP.

The co-digestion of pretreated sugarcane lignocelluloses with dairy cow manure (DCM) as a bioenergy production and waste management strategy, for intensive livestock farms located in sugarcane regions, was investigated. Ammonia fiber expansion (AFEX) increased the nitrogen content and accelerated the biodegradability of sugarcane bagasse (SCB) and cane leaf matter (CLM) through the cleavage of lignin carbohydrate crosslinks, resulting in the highest specific methane yields (292-299 L CH4/kg VSadded), biogas methane content (57-59% v/v) and biodegradation rates, with or without co-digestion with DCM. To obtain comparable methane yields, untreated and steam exploded (StEx) SCB and CLM had to be co-digested with DCM, at mass ratios providing initial C/N ratios in the range of 18 to 35. Co-digestion with DCM improved the nutrient content of the solid digestates, providing digestates that could be used as biofertilizer to replace CLM that is removed from sugarcane fields during green harvesting.

This report outlines the recent scale-up of AFEX pretreatment from the laboratory to pilot scale. Sugar yields were improved by 19 and 15% for glucose and xylose, respectively. Further improvement was achieved when scaling up the hydrolysis and fermentation of AFEX-treated corn stover to 2500 L working volume. Benchmarking was performed using CTec 3 and HTec 3 enzymes along with Zymomonas mobilis 8b. Subsequently, the seed train was modified to use a hydrolysate-based medium as a replacement for pure sugars and nutrients. Fermentation performance was comparable following this change. Economic analysis showed a 19% reduction in MESP for the new process over the previous benchmark process.

One of the key challenges facing lignin conversion to fuels and chemicals is related to the level of carbohydrate and ash impurities found in extracted lignin. Structural modifications of lignin may also occur as a result of biomass pretreatment and harsh lignin extraction protocols. Extractive-Ammonia (EA) is a new pretreatment technology that uses liquid ammonia to cleave lignin–carbohydrate complexes, decrystallize cellulose, solubilize lignin, and selectively extract lignin from lignocellulosic biomass, enabling better utilization of both lignin and carbohydrate components in a biorefinery. The EA-based biorefinery produces two different lignin-rich streams, with different properties, that could potentially be upgraded to fuels and chemicals using green processes. In this work, a water/ethanol-based fractionation method was developed to enrich the ammonia-soluble extractives, resulting in a major product stream containing 92% lignin. Detailed characterization of the various streams resulting from EA treatment, including compositional analysis, structural characterization by nuclear magnetic resonance (NMR) spectrometry, elemental analysis, molecular weight analysis, and thermo-gravimetric analysis provides a broad evaluation of the EA-derived lignin product stream structures and properties, assessing their potential for commercial applications. In summary, EA-derived lignins preserve much of lignin's functionality, including the sensitive β-aryl ether units. Nitrogen incorporation was observed in the lignin-rich streams, notably due to the presence of hydroxycinnamoyl amides formed during ammonia pretreatment.

A new liquid ammonia pretreatment methodology called Extractive Ammonia (EA) was developed to simultaneously convert native crystalline cellulose Iβ (CI) to a highly digestible cellulose IIII (CIII) allomorph and selectively extract up to ∼45% of the lignin from lignocellulosic biomass with near-quantitative retention of all polysaccharides. EA pretreated corn stover yielded a higher fermentable sugar yield compared to the older Ammonia Fiber Expansion (AFEX) process while using 60% lower enzyme loading. The EA process preserves extracted lignin functionalities, offering the potential to co-produce lignin-derived fuels and chemicals in the biorefinery. The single-stage EA fractionation process achieves high biofuel yields (18.2 kg ethanol per 100 kg untreated corn stover, dry weight basis), comparable to those achieved using ionic liquid pretreatments. The EA process achieves these ethanol yields at industrially-relevant conditions using low enzyme loading (7.5 mg protein per g glucan) and high solids loading (8% glucan, w/v).

In response to growing concerns about energy security, environmental sustainability and societal sustainability, cellulosic biomass refining technologies have been extensively developed in recent years. However, these technologies are not yet fully commercialized. High capital cost and high enzyme cost are two major bottlenecks. Capital cost and operating cost (excluding 33% feedstock cost) account for 34% and 33%, respectively, of the total biofuel production cost with enzyme cost alone representing about 47% of the operating cost. Therefore, reducing both capital cost and enzyme cost is imperative. Over the past eight years, with the support from US Department of Energy Great Lakes Bioenergy Research Center (GLBRC), we greatly improved our AFEX™ (Trade mark of MBI, International (Lansing, Michigan)) (Ammonia Fiber Expansion)-related processing technologies, leading to a 66% reduction in enzyme loading (current enzyme loading is as low as 7.5 mg protein per g glucan) and a 129% enhancement in ethanol volumetric productivity (>56% reduction in capital cost for enzymatic hydrolysis and fermentation).

Lignin is the second most abundant biopolymer on the earth, yet its utilization for fungible products is complicated by its recalcitrant nature and remains a major challenge for sustainable lignocellulosic biorefineries. In this study, we used a systems biology approach to reveal the carbon utilization pattern and lignin degradation mechanisms in a unique lignin-utilizing Pseudomonas putida strain (A514). The mechanistic study further guided the design of three functional modules to enable a consolidated lignin bioconversion route. First, P. putida A514 mobilized a dye peroxidase-based enzymatic system for lignin depolymerization. This system could be enhanced by overexpressing a secreted multifunctional dye peroxidase to promote a two-fold enhancement of cell growth on insoluble kraft lignin. Second, A514 employed a variety of peripheral and central catabolism pathways to metabolize aromatic compounds, which can be optimized by overexpressing key enzymes. Third, the β-oxidation of fatty acid was up-regulated, whereas fatty acid synthesis was down-regulated when A514 was grown on lignin and vanillic acid. Therefore, the functional module for polyhydroxyalkanoate (PHA) production was designed to rechannel β-oxidation products. As a result, PHA content reached 73% per cell dry weight (CDW). Further integrating the three functional modules enhanced the production of PHA from kraft lignin and biorefinery waste. Thus, this study elucidated lignin conversion mechanisms in bacteria with potential industrial implications and laid out the concept for engineering a consolidated lignin conversion route.

Current models of cellulosic biofuel systems require that the delivered price of the cellulosic biomass feedstock be kept low. Thus the predicted biorefinery size is relatively small, limiting potential economies of scale. However, it is actually the ultimate selling price of the biofuel that largely determines market penetration. We relaxed the constraint of low delivered feedstock price and explored the resulting effects on biofuel price, biofuel volume produced, and global warming impact (GWI). Feedstock price greatly affects the feedstock supply chains that may develop. Increased feedstock price does not affect the final ethanol selling price very much, but higher feedstock prices greatly increase the amount of ethanol produced. Farmers will supply much more cellulosic biomass at higher feedstock prices, leading to shorter transportation distances with reduced transportation costs and enabling larger biorefineries with improved economies of scale, thereby reducing the ethanol selling price. The cellulosic feedstock supply chain systems were studied as a function of feedstock prices by determining potential feedstock supply clusters and the maximum capacity of cellulosic biorefineries across the United States. Supply clusters were determined by minimizing costs associated with ethanol production. The analysis is based on county-level cellulosic feedstock production data projected in the US Billion-Ton Update report. Each biomass supply cluster is unique in terms of local and regional characteristics (e.g. area, feedstock types), biorefinery capacity, ethanol selling price, and GWI. Very large-scale biorefineries (≥20 000 dry Mg day−1) may be feasible in some regions. © 2015 Society of Chemical Industry and John Wiley & Sons, Ltd