In vitro gas production as a surrogate measure of the fermentability of cellulosic biomass to ethanol

Current methods for measuring ethanol yields from lignocellulosic biomass are relatively slow and are not well geared for analyzing large numbers of samples generated by feedstock management and breeding research. The objective of this study was to determine if an in vitro ruminal fermentation assay used in forage quality research was predictive of results obtained using a conventional biomass-to-ethanol conversion assay. In the conventional assay, herbaceous biomass samples were converted to ethanol by Saccharomyces cerevisiae cultures in the presence of cellulase enzymes. Cultures were grown in sealed serum bottles and gas production monitored by measuring increasing head space pressure. Gas accumulation as calculated from the pressure measurements was highly correlated (r2\u3e0.9) with ethanol production measured by gas chromatography at 24 h or 7 days. The same feedstocks were also analyzed by in vitro ruminal digestion, as also measured by gas accumulation. Good correlations (r2∼0.63–0.82) were observed between ethanol production during simultaneous saccharification and fermentation and gas accumulation in parallel in vitro ruminal fermentations. Because the in vitro ruminal fermentation assay can be performed without sterilization of the medium and does not require aseptic conditions, this assay may be useful for biomass feedstock agronomic and breeding research

Switchgrass Biomass Composition Altered by Six Generations of Divergent Breeding for Digestibility

Biomass composition of switchgrass (Panicum virgatum L.) can affect its utilization by ruminants and its conversion to liquid fuels in a biorefinery. The objective of this study was to evaluate the effects of six generations of divergent breeding for forage in vitro dry matter digestibility (IVDMD) on switchgrass biomass composition, forage quality traits, and ethanol yield. Initially there was one cycle of selection for both low (C-1) and high IVDMD (C1 = cv. Trailblazer), followed by four additional breeding cycles for high IVDMD. In cycles 4 and 5, winter survival was included as a selection criterion because of decreased winter survival of the C3 population. The experimental populations that were produced by these breeding generations and nine half-sib families from cycle 5 were evaluated for two post-establishment years at the research station in eastern Nebraska, where all the breeding work was conducted. The six breeding generations resulted in significant differences among the populations for all the 28 cell wall and non-cell-wall composition variables measured, forage quality, and ethanol yield traits measured except for total biomass C, cell wall concentration, soluble glucose, and etherified ferulates. These traits included all cell wall and nonstructural carbohydrates. Breeding for the heritable complex trait IVDMD affected a large number of plant biomass characteristics and also adversely affected plant biomass yield and winter survival

Temporal and Spatial Variation in Switchgrass Biomass Composition and Theoretical Ethanol Yield

Information on temporal and spatial variation in switchgrass (Panicum virgatum L.) biomass composition as it affects ethanol yield (L Mg–1) at a biorefinery and ethanol production (L ha–1) at the field-scale has previously not been available. Switchgrass biomass samples were collected from a regional, on-farm trial and biomass composition was determined using newly developed near-infrared reflectance spectroscopy (NIRS) prediction equations and theoretical ethanol yield (100% conversion efficiency) was calculated. Total hexose (cell wall polysaccharides and soluble sugars) concentration ranged from 342 to 398 g kg–1 while pentose (arabinose and xylose) concentration ranged from 216 to 245 g kg–1 across fields. Theoretical ethanol yield varied significantly by year and field, with 5 yr means ranging from 381 to 430 L Mg–1. Total theoretical ethanol production ranged from 1749 to 3691 L ha–1 across fields. Variability (coefficient of variation) within established switchgrass fields ranged from 1 to 4% for theoretical ethanol yield (L Mg–1) and 14 to 38% for theoretical ethanol production (L ha–1). Most fields showed a lack of spatial consistency across harvest years for theoretical ethanol yield or total theoretical ethanol production. Switchgrass biomass composition from farmer fields can be expected to have significant annual and field-to-field variation in a production region, and this variation will significantly affect ethanol or other liquid fuel yields per ton or hectare. Cellulosic biorefineries will need to consider this potential variation in biofuel yields when developing their business plans

Ethanol yields and cell wall properties in divergently bred switchgrass genotypes

Genetic modification of herbaceous plant cell walls to increase biofuels yields is a primary bioenergy research goal. Using two switchgrass populations developed by divergent breeding for ruminant digestibility, the contributions of several wall-related factors to ethanol yields was evaluated. Field grown low lignin plants significantly out yielded high lignin plants for conversion to ethanol by 39.1% and extraction of xylans by 12%. However, across all plants analyzed, greater than 50% of the variation in ethanol yields was attributable to changes in tissue and cell wall architecture, and responses of stem biomass to diluteacid pretreatment. Although lignin levels were lower in the most efficiently converted genotypes, no apparent correlation were seen in the lignin monomer G/S ratios. Plants with higher ethanol yields were associated with an apparent decrease in the lignification of the cortical sclerenchyma, and a marked decrease in the granularity of the cell walls following dilute-acid pretreatment

Ethanol yields and cell wall properties in divergently bred switchgrass genotypes

Genetic modification of herbaceous plant cell walls to increase biofuels yields is a primary bioenergy research goal. Using two switchgrass populations developed by divergent breeding for ruminant digestibility, the contributions of several wall-related factors to ethanol yields was evaluated. Field grown low lignin plants significantly out yielded high lignin plants for conversion to ethanol by 39.1% and extraction of xylans by 12%. However, across all plants analyzed, greater than 50% of the variation in ethanol yields was attributable to changes in tissue and cell wall architecture, and responses of stem biomass to diluteacid pretreatment. Although lignin levels were lower in the most efficiently converted genotypes, no apparent correlation were seen in the lignin monomer G/S ratios. Plants with higher ethanol yields were associated with an apparent decrease in the lignification of the cortical sclerenchyma, and a marked decrease in the granularity of the cell walls following dilute-acid pretreatment

Comparison of Cellulosic Ethanol Yields from Midwestern Maize and Reconstructed Tallgrass Prairie Systems Managed for Bioenergy

Maize- and prairie-based systems were investigated as cellulosic feedstocks by conducting a 9 ha side-by-side comparison on fertile soils in the Midwestern United States. Maize was grown continuously with adequate fertilization over years both with and without a winter rye cover crop, and the 31-species reconstructed prairie was grown with and without spring nitrogen fertilization. Both maize stover and prairie biomass were harvested in the fall. We compared amounts of cellulosic biomass produced and harvested, carbohydrate contents as measured by both dietary and detergent methods, and estimated cellulosic ethanol yields per hectare. From 2009–2013, the cropping system with the largest non-grain biomass yield was fertilized prairie, averaging 10.4 Mg ha−1 year−1 aboveground biomass with average harvest removals of 7.8 Mg ha−1 year−1. The unfertilized prairie produced 7.4 Mg ha−1 year−1 aboveground biomass, with average harvests of 5.3 Mg ha−1 year−1. Lowest cellulosic (non-grain) biomass harvests were obtained from continuous maize systems, averaging 3.5 Mg ha−1 year−1 when grown with, and 3.7 Mg ha−1 year−1 when grown without a winter rye cover crop, respectively. Unfertilized prairie biomass and maize stover had equivalent dietary-determined potential biomass ethanol yields at 330 g ethanol kg−1 dry biomass, but fertilized prairie was lower at 315. The detergent method did not accurately capture these differences. Over the five-year period of the experiment, unfertilized and fertilized prairie systems averaged 810 and 1,790 L potential cellulosic ethanol ha−1 year−1 more than the maize systems, respectively. Differences in harvested biomass accounted for \u3e90 % of ethanol yield variation

Bioconversion of Pelletized Big Bluestem, Switchgrass, and Low-Diversity Grass Mixtures Into Sugars and Bioethanol

Three crops of warm-season grasses are being developed for biomass production on northern rain-fed marginal farmland: big bluestem (BBS), switchgrass (SG), and a low diversity mixture of grasses (LDM). In this study, biomass harvested from established fields were compared for pelletization and subsequent conversion to sugars and ethanol. Each biomass was successfully pelletized to similar bulk densities without adding a binder at a commercial feed operation. Pelletizing increased the bulk density by 407% on average and was equally effective on all three biomass samples (528–554 kg/m3). Chemical analysis of the samples indicated that glucan and xylan contents were slightly reduced during pelletizing (by 23 and 16 g/kg, respectively), as well as theoretical ethanol yields, which are based upon total carbohydrate contents. Pellets and milled straws were pre-treated with either liquid hot-water or low-moisture ammonium hydroxide (LMA) and subsequently hydrolyzed with cellulases. Glucose and total sugar yields were similar for non-pellets and pellets using either pre-treatment; carbohydrates present in pellets were more efficiently recovered compared to non-pellets. LMA pretreated samples were separately hydrolyzed and fermented to ethanol using Scheffersomyces stipitis yeast. Hydrolysis recovered 69.7–76.8% of the glucose and 66.5–73.3% of the xylose across all samples. Glucose yields were 251–279 g/kg, db and were significantly lower for SG as compared to the other biomass samples. Recovered sugars were fermented to ethanol at 77.7–86.7% of theoretical yield. Final ethanol yields (245.9–275.5 L/Mg, db) were similar for all of the grasses and estimated to equate to production levels for BBS, LDM, and SG of 1,952, 2,586, and 2,636 l of ethanol per ha, respectively