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Concept: Biofuel


BACKGROUND: The recalcitrance of lignocellulosic cell wall biomass to deconstruction varies greatly in angiosperms, yet the source of this variation remains unclear. Here, in eight genotypes of short rotation coppice willow (Salix sp.) variability of the reaction wood (RW) response and the impact of this variation on cell wall recalcitrance to enzymatic saccharification was considered. RESULTS: A pot trial was designed to test if the ‘RW response’ varies between willow genotypes and contributes to the differences observed in cell wall recalcitrance to enzymatic saccharification in field-grown trees. Biomass composition was measured via wet chemistry and used with glucose release yields from enzymatic saccharification to determine cell wall recalcitrance. The levels of glucose release found for pot-grown control trees showed no significant correlation with glucose release from mature field-grown trees. However, when a RW phenotype was induced in pot-grown trees, glucose release was strongly correlated with that for mature field-grown trees. Field studies revealed a 5-fold increase in glucose release from a genotype grown at a site exposed to high wind speeds (a potentially high RW inducing environment) when compared with the same genotype grown at a more sheltered site. CONCLUSIONS: Our findings provide evidence for a new concept concerning variation in the recalcitrance to enzymatic hydrolysis of the stem biomass of different, field-grown willow genotypes (and potentially other angiosperms). Specifically, that genotypic differences in the ability to produce a response to RW inducing conditions (a ‘RW response’) indicate that this RW response is a primary determinant of the variation observed in cell wall glucan accessibility. The identification of the importance of this RW response trait in willows, is likely to be valuable in selective breeding strategies in willow (and other angiosperm) biofuel crops and, with further work to dissect the nature of RW variation, could provide novel targets for genetic modification for improved biofuel feedstocks.

Concepts: DNA, Gene, Genetics, Genotype, Evolution, Enzyme, Biofuel, Willow


Miscanthus sacchariflorus is a fast-growing C(4) perennial grass that can naturally hybridize with M. sinensis to produce interspecific hybrids, such as the sterile triploid M.× giganteus. The creation of such hybrids is essential for the rapid domestication of this novel bioenergy crop. However, progress has been hindered by poor understanding of the environmental cues promoting floral transition in M. sacchariflorus, which flowers less readily than M. sinensis. The purpose of this work was to identify the flowering requirements of M. sacchariflorus genotypes in order to expedite the introduction of new germplasm optimized to different environments. Six M. sacchariflorus accessions collected from a range of latitudes were grown under controlled photoperiod and temperature conditions, and flowering, biomass, and morphological phenotypic data were captured. Results indicated that M. sacchariflorus, irrespective of origin, is a quantitative short-day plant. Flowering under static long days (15.3h daylength), compared with shorter photoperiods, was delayed by an average 61 d, with an average associated increase of 52% of above-ground biomass (DM plant(-1)). Timing of floral initiation occurred between photoperiods of 14.2h and 12.1h, and accumulated temperatures of 553-1157 °C above a base temperature of 10 °C. Miscanthus sacchariflorus flowering phenology closely resembles that of Sorghum and Saccharum, indicating potentially similar floral pathways and suggesting that determination of the underlying genetic mechanisms will be facilitated by the syntenic relationships existing between these important C(4) grasses.

Concepts: Evolution, Poaceae, Maize, Flower, Biofuel, Photoperiodism, Biomass, Miscanthus giganteus


The considerable increase in biodiesel production worldwide in the last 5 years resulted in a stoichiometric increased coproduction of crude glycerol. As an excess of crude glycerol has been produced, its value on market was reduced and it is becoming a “waste-stream” instead of a valuable “coproduct”. The development of biorefineries, i.e. production of chemicals and power integrated with conversion processes of biomass into biofuels, has been singled out as a way to achieve economically viable production chains, valorize residues and coproducts, and reduce industrial waste disposal. In this sense, several alternatives aimed at the use of crude glycerol to produce fuels and chemicals by microbial fermentation have been evaluated. This review summarizes different strategies employed to produce biofuels and chemicals (1,3-propanediol, 2,3-butanediol, ethanol, n-butanol, organic acids, polyols and others) by microbial fermentation of glycerol. Initially, the industrial use of each chemical is briefly presented; then we systematically summarize and discuss the different strategies to produce each chemical, including selection and genetic engineering of producers, and optimization of process conditions to improve yield and productivity. Finally, the impact of the developments obtained until now are placed in perspective and opportunities and challenges for using crude glycerol to the development of biodiesel-based biorefineries are considered. In conclusion, the microbial fermentation of glycerol represents a remarkable alternative to add value to the biodiesel production chain helping the development of biorefineries, which will allow this biofuel to be more competitive.

Concepts: Carbon dioxide, Ethanol, Petroleum, Anaerobic digestion, Biofuel, Glycerol, Biomass, Biodiesel


Bioenergy will be one component of a suite of alternatives to fossil fuels. Effective conversion of biomass to energy will require the careful pairing of advanced conversion technologies with biomass feedstocks optimized for the purpose. Lignocellulosic biomass can be converted to useful energy products via two distinct pathways: enzymatic or thermochemical conversion. The thermochemical pathways are reviewed and potential biotechnology or breeding targets to improve feedstocks for pyrolysis, gasification, and combustion are identified. Biomass traits influencing the effectiveness of the thermochemical process (cell wall composition, mineral and moisture content) differ from those important for enzymatic conversion and so properties are discussed in the language of biologists (biochemical analysis) as well as that of engineers (proximate and ultimate analysis). We discuss the genetic control, potential environmental influence, and consequences of modification of these traits. Improving feedstocks for thermochemical conversion can be accomplished by the optimization of lignin levels, and the reduction of ash and moisture content. We suggest that ultimate analysis and associated properties such as H:C, O:C, and heating value might be more amenable than traditional biochemical analysis to the high-throughput necessary for the phenotyping of large plant populations. Expanding our knowledge of these biomass traits will play a critical role in the utilization of biomass for energy production globally, and add to our understanding of how plants tailor their composition with their environment.

Concepts: Metabolism, Hydrogen, Petroleum, Cell wall, Biofuel, Combustion, Biomass, Pyrolysis


Fuel ethanol production from sustainable and largely abundant agro-residues such as sugarcane bagasse (SB) provides long term, geopolitical and strategic benefits. Pretreatment of SB is an inevitable process for improved saccharification of cell wall carbohydrates. Recently, ammonium hydroxide-based pretreatment technologies have gained significance as an effective and economical pretreatment strategy. We hypothesized that soaking in concentrated aqueous ammonia-mediated thermochemical pretreatment (SCAA) would overcome the native recalcitrance of SB by enhancing cellulase accessibility of the embedded holocellulosic microfibrils.

Concepts: Ammonia, Ethanol, Cellulose, Hydrolysis, Biofuel, Ethanol fuel, Sugarcane, Bagasse


In the US Corn Belt, a recent doubling in commodity prices has created incentives for landowners to convert grassland to corn and soybean cropping. Here, we use land cover data from the National Agricultural Statistics Service Cropland Data Layer to assess grassland conversion from 2006 to 2011 in the Western Corn Belt (WCB): five states including North Dakota, South Dakota, Nebraska, Minnesota, and Iowa. Our analysis identifies areas with elevated rates of grass-to-corn/soy conversion (1.0-5.4% annually). Across the WCB, we found a net decline in grass-dominated land cover totaling nearly 530,000 ha. With respect to agronomic attributes of lands undergoing grassland conversion, corn/soy production is expanding onto marginal lands characterized by high erosion risk and vulnerability to drought. Grassland conversion is also concentrated in close proximity to wetlands, posing a threat to waterfowl breeding in the Prairie Pothole Region. Longer-term land cover trends from North Dakota and Iowa indicate that recent grassland conversion represents a persistent shift in land use rather than short-term variability in crop rotation patterns. Our results show that the WCB is rapidly moving down a pathway of increased corn and soybean cultivation. As a result, the window of opportunity for realizing the benefits of a biofuel industry based on perennial bioenergy crops, rather than corn ethanol and soy biodiesel, may be closing in the WCB.

Concepts: Agriculture, Native Americans in the United States, Biofuel, Ethanol fuel, Minnesota, American Bison, South Dakota, National Agricultural Statistics Service


Biofuels are the most immediate, practical solution for mitigating dependence on fossil hydrocarbons, but current biofuels (alcohols and biodiesels) require significant downstream processing and are not fully compatible with modern, mass-market internal combustion engines. Rather, the ideal biofuels are structurally and chemically identical to the fossil fuels they seek to replace (i.e., aliphatic n- and iso-alkanes and -alkenes of various chain lengths). Here we report on production of such petroleum-replica hydrocarbons in Escherichia coli. The activity of the fatty acid (FA) reductase complex from Photorhabdus luminescens was coupled with aldehyde decarbonylase from Nostoc punctiforme to use free FAs as substrates for alkane biosynthesis. This combination of genes enabled rational alterations to hydrocarbon chain length (Cn) and the production of branched alkanes through upstream genetic and exogenous manipulations of the FA pool. Genetic components for targeted manipulation of the FA pool included expression of a thioesterase from Cinnamomum camphora (camphor) to alter alkane Cn and expression of the branched-chain α-keto acid dehydrogenase complex and β-keto acyl-acyl carrier protein synthase III from Bacillus subtilis to synthesize branched (iso-) alkanes. Rather than simply reconstituting existing metabolic routes to alkane production found in nature, these results demonstrate the ability to design and implement artificial molecular pathways for the production of renewable, industrially relevant fuel molecules.

Concepts: Fatty acid, Ethanol, Petroleum, Hydrocarbon, Alkane, Biofuel, Methane, Internal combustion engine


Prospecting macroalgae (seaweeds) as feedstocks for bioconversion into biofuels and commodity chemical compounds is limited primarily by the availability of tractable microorganisms that can metabolize alginate polysaccharides. Here, we present the discovery of a 36-kilo-base pair DNA fragment from Vibrio splendidus encoding enzymes for alginate transport and metabolism. The genomic integration of this ensemble, together with an engineered system for extracellular alginate depolymerization, generated a microbial platform that can simultaneously degrade, uptake, and metabolize alginate. When further engineered for ethanol synthesis, this platform enables bioethanol production directly from macroalgae via a consolidated process, achieving a titer of 4.7% volume/volume and a yield of 0.281 weight ethanol/weight dry macroalgae (equivalent to ~80% of the maximum theoretical yield from the sugar composition in macroalgae).

Concepts: Bacteria, Metabolism, Enzyme, Glucose, Starch, Biofuel, Ethanol fuel, Algaculture


Recent energy security strategies, investment opportunities and energy policies have led to an escalation in biofuel consumption at the expenses of food crops and pastureland. To evaluate the important impacts of biofuels on food security, the food-energy nexus needs to be investigated in the context of its linkages with the overall human appropriation of land and water resources. Here we provide a global assessment of biofuel crop production, reconstruct global patterns of biofuel crop/oil trade and determine the associated displacement of water and land use. We find that bioethanol is mostly produced with domestic crops while 36% of biodiesel consumption relies on international trade, mainly from Southeast Asia. Altogether, biofuels rely on about 2-3% of the global water and land used for agriculture, which could feed about 30% of the malnourished population. We evaluate the food-energy tradeoff and the impact an increased reliance on biofuel would have on the number of people the planet can feed.

Concepts: Human, Agriculture, Poverty, Ethanol, Food security, Biofuel, World population, Biodiesel


Growth in biofuel production, which is meant to reduce greenhouse gas (GHG) emissions and fossil en-ergy demand, is increasingly seen as a threat to food supply and natural habitats. Using photovoltaics (PV) to directly convert solar radiation into electricity for battery electric vehicles (BEVs) is an alterna-tive to photosynthesis, which suffers from a very low energy conversion efficiency. Assessments need to be spatially explicit, since solar insolation and crop yields vary widely between locations. This paper therefore compares direct land use, life cycle GHG emissions and fossil fuel requirements of five differ-ent sun-to-wheels conversion pathways for every county in the contiguous U.S.: Ethanol from corn or switchgrass for internal combustion vehicles (ICVs), electricity from corn or switchgrass for BEVs, and PV electricity for BEVs. Even the most land-use efficient biomass-based pathway (i.e., switchgrass bio-electricity in U.S. counties with hypothetical crop yields of over 24 tonnes/ha) requires 29 times more land than the PV-based alternative in the same locations. PV BEV systems also have the lowest life cy-cle GHG emissions throughout the U.S. and the lowest fossil fuel inputs, except for locations with hypothetical switchgrass yields of 16 or more tonnes/ha. Including indirect land use effects further strengthens the case for PV.

Concepts: Carbon dioxide, Solar cell, Photovoltaics, Biofuel, Fossil fuel, Greenhouse gas, Automobile, Internal combustion engine