Cellulosic Ethanol Conversion Research Articles

Cover Image, Volume 16, Issue 3

AbstractThe cover image is based on the Research Article Techno‐economic analysis of cellulosic ethanol conversion to fuel and chemicals by Steven Phillips et al., https://doi.org/10.1002/bbb.2346. image

Techno‐economic analysis of cellulosic ethanol conversion to fuel and chemicals

AbstractIn this paper, we describe multiple scenarios to assess the techno‐economic results of producing hydrocarbon fuels from corn stover‐derived ethanol and the impact of co‐producing higher‐value, ethanol‐derived n‐butanol and 1,3‐butadiene to improve economic feasibility. In our baseline process, the stover‐derived lignin is burned for its energy content to raise process steam and electricity. We evaluated an alternative approach based on the same stover‐to‐ethanol process, but instead of burning residual lignin to produce heat and electricity, it was fed to a hydrothermal liquefaction process and converted into hydrocarbon fuel, while the ethanol from fermentation is used to make only butanol or butadiene. Our results indicate that the hydrothermal liquefaction pathways have significant cost advantages over the pathways that burn the lignin. For comparison purposes, two more scenarios were evaluated based on the assumption that ethanol was purchased at market prices and converted to butanol or butadiene exclusively. This comparison evaluates the economics for making higher‐valued chemicals which, in the absence of incentives, would be the most profitable approaches in our study. The results for these evaluations indicated that the ethanol‐to‐butanol economics were generally favorable and achieved or exceeded the 10% internal rate of return target. The butadiene process had less favorable economic performance in some years when the market price for butadiene was less than the selling price needed to achieve the internal rate of return. Economic incentives for low‐carbon chemicals and fuels were not considered in our analysis. © 2022 Battelle Memorial Institute. Biofuels, Bioproducts and Biorefining published by Society of Industrial Chemistry and John Wiley & Sons Ltd.

Biomass yield and quality of the Miscanthus × giganteus in northern China

AbstractMiscanthus × giganteus has attracted much attention and occupied a significant niche as a bioenergy crop to meet the world's energy demand and in the fight against climate change. In this study, a field experiment was conducted to evaluate the yield and biomass characteristics of M. × giganteus over a period of three years from 2013 to 2015 in Beijing, China. M. × giganteus plants showed higher tiller density and litter leaf weight in the third year, whereas no significant difference in plant height was observed among the years. In contrast, a larger number of nodes per plant were recorded in the first year. Biomass yield potential was the highest in the third year (42.3 t ha−1). In addition, plant height (r = .594, p = .042), tiller density (r = .854, p < .01), and number of nodes per plant (r = ‐.589, p = .044) were significantly correlated with biomass yield. Contents of cellulose and lignin were higher in stems, while that of hemicellulose and ash were higher in leaves. M. × giganteus biomass may be used for cellulosic ethanol conversion due to high cellulose and hemicellulose contents and low lignin content and for direct combustion and co‐combustion due to high calorific value (18.5 MJ kg−1). To conclude, our findings indicate that M. × giganteus can be cultivated in northern China as an excellent herbaceous bioenergy crop. Rational utilization of the agronomic characteristics would be helpful to screening and breeding bioenergy grass for promoting bioenergy production in China.

Genome-wide analysis and characterization of GRAS family in switchgrass

ABSTRACT Panicum virgatum, a model plant of cellulosic ethanol conversion, not only has high large biomass and strong adaptability to soil, but also grows well in marginal soil and has the advantage of improving saline-alkali soil. GRAS transcription factor gene family play important roles in individual environment adaption, and these vital functions has been proved in several plants, however, the research of GRAS in the development of switchgrass (Panicum virgatum) were limited. A comprehensive study was investigated to explore the relationship between GRAS gene family and resistance. According to the phylogenetic analysis, a total of 144 GRAS genes were identified and renamed which were classified into eight subfamilies. Chromosome distribution, tandem and segmental repeats analysis indicated that gene duplication events contributed a lot to the expansion of GRAS genes in the switchgrass genome. Sixty-six GRAS genes in switchgrass were identified as having orthologous genes with rice through gene duplication analysis. Most of these GRAS genes contained zero or one intron, and closely related genes in evolution shared similar motif composition. Interaction networks were analyzed including DELLA and ten interaction proteins that were primarily involved in gibberellin acid mediated signaling. Notably, online analysis indicated that the promoter regions of the identified PvGRAS genes contained many cis-elements including light responsive elements, suggesting that PvGRAS might involve in light signal cross-talking. This work provides key insights into resistance and bioavailability in switchgrass and would be helpful to further study the function of GRAS and GRAS-mediated signal transduction pathways.

Improved cellulosic ethanol production from corn stover with a low cellulase input using a β-glucosidase-producing yeast following a dry biorefining process.

A low-cost and sustainable cellulosic ethanol production is vital for fermentation-based industrial applications. Reducing the expenses of cellulose-deconstruction enzymes is one of the significant challenges to economic cellulose-to-ethanol conversion. Here, we report the improved ethanol production from corn stover after dry biorefining using a natural β-glucosidase-producing strain Clavispora NRRL Y-50464 with a low cellulase dose of 5mg protein/g glucan under separate enzymatic hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) conditions. Strain Clavispora NRRL Y-50464 exhibited a superior ethanol fermentation performance over Saccharomyces cerevisiae DQ1 under both conditions. It produced an ethanol titer of 38.1g/L within 96h at a conversion efficiency of 55.5% with 25% solids loading (w/w) via SSF without addition of extra β-glucosidase supplement. Improved performance of Y-50464 on a bioreactor with a helical stirring apparatus confirmed its advantage over the conventional bioreactors originally designed for liquid fermentations in cellulosic ethanol conversion by SSF. The results of this study suggested that the strain Clavispora NRRL Y-50464 has a potential as a candidate for lower-cost cellulosic ethanol production from lignocellulosic materials.

Consolidated bioprocessing for cellulosic ethanol conversion by cellulase–xylanase cell-surfaced yeast consortium

Consolidated bioprocessing (CBP) strategy was developed to construct a cell-surface displayed consortium for heterologously expressing functional lignocellulytic enzymes. The reaction system composed of two engineered yeast strains: Y5/XynII-XylA (co-displaying two types of xylanases) and Y5/EG-CBH-BGL (co-displaying three types of cellulases). The immobilization of recombinant fusion proteins and their cell-surface accessibility of were analyzed by flow cytometry and immunofluorescence. The feasibility of consolidated bioprocessing by using pretreated corn stover (CS) as substrate for direct bioconversion was further investigated, and the synergistic activity and proximity effect between cellulases and xylanases on lignocelluloses degradation were also discussed in this work. Without any commercial enzyme addition, the combined yeast consortium produced 1.61 g/L ethanol which achieved 64.7% of the theoretical ethanol yield during 144 h from steam-exploded CS. The results indicated that the assembly of cellulases and xylanases using a synthetic consortium capable of combined displaying lignocellulytic enzymes is a promising approach for simultaneous saccharification and fermentation to ethanol from lignocellulosic biomass.

Lifecycle economic analysis of biofuels: Accounting for economic substitution in policy assessment

This paper develops a lifecycle economic analysis (LCEA) model that integrates endogenous input substitution into the standard lifecycle analysis (LCA) of biofuel that typically assumes fixed-proportions production. We use the LCEA model to examine impacts of a pure carbon tax and a revenue-neutral tax-subsidy policy on lifecycle greenhouse gas emissions from cellulosic ethanol using forest residues as feedstock in Washington State. In a model allowing for input substitution in the cellulosic ethanol feedstock, conversion, and transportation process, we consider energy source substitution (woody biomass for coal in the cellulosic ethanol conversion plant and biodiesel for diesel in feedstock production and feedstock and ethanol transportation) as well as substitution of capital and labor for energy in all stages of the lifecycle. We find that ignoring endogenous input substitution by using standard LCA leads to substantial underestimation of the impact of carbon tax policies on carbon emissions. Both tax policies can substantially reduce carbon emissions by inducing substitution among inputs. The revenue-neutral tax-subsidy policy reduces emissions more effectively than the carbon tax policy for carbon tax rates currently in place throughout most of the world. It stimulates substitution of woody biomass for coal and biodiesel for diesel at much lower tax rates when accompanied by corresponding subsidies for reduced emissions from renewable sources.

RNA-seq Analysis of Overexpressing Ovine AANAT Gene of Melatonin Biosynthesis in Switchgrass.

Melatonin serves important functions in the promotion of growth and anti-stress regulation by efficient radical scavenging and regulation of antioxidant enzyme activity in various plants. To investigate its regulatory roles and metabolism pathways, the transcriptomic profile of overexpressing the ovine arylalkylamine N-acetyltransferase (oAANAT) gene, encoding the penultimate enzyme in melatonin biosynthesis, was compared with empty vector control using RNA-seq in switchgrass, a model plant of cellulosic ethanol conversion. The 85.22 million high quality reads that were assembled into 135,684 unigenes were generated by Illumina sequencing for transgenic oAANAT switchgrass with an average sequence length of 716 bp. A total of 946 differentially expression genes in transgenic line comparing to control switchgrass, including 737 up-regulated and 209 down-regulated genes, were mainly enriched with two main functional patterns of melatonin identifying by gene ontology analysis: the growth regulator and stress tolerance. Furthermore, KEGG maps indicated that the biosynthetic pathways of secondary metabolite (phenylpropanoids, flavonoids, steroids, stilbenoid, diarylheptanoid, and gingerol) and signaling pathways (MAPK signaling pathway, estrogen signaling pathway) were involved in melatonin metabolism. This study substantially expands the transcriptome information for switchgrass and provides valuable clues for identifying candidate genes involved in melatonin biosynthesis and elucidating the mechanism of melatonin metabolism.

목질계 셀룰로오스 에탄올 생산공정에서 전처리과정의 설계

A pretreatment process of cellulose decomposition to a monosaccharide plays an important role in the cellulosic ethanol production using the lignocellulosic biomass. In this study, a cellulosic ethanol was produced by using acidic hydrolysis and enzymatic saccharification process from the lignocellulosic biomass such as rice straw, sawdust, copying paper and newspaper. Three different pretreatment processes were compared; the acidic hydrolysis (100 ℃, 1 h) using 10~30 wt% of sulfuric acid, the enzymatic saccharification (30 min) using celluclast (55 ℃, pH = 5.0), AMG (60 ℃, pH = 4.5), and spirizyme (60 ℃, pH = 4.2) and also the hybrid process (enzymatic saccharification after acidic hydrolysis). The yield of cellulosic ethanol conversion with those pretreatment processes were obtained as the following order : hybrid process > acidic hydrolysis > enzymatic saccharification. The optimum fermentation time was proven to be two days in this work. The yield of cellulosic ethanol conversion using celluclast after the acidic hydrolysis with 20 wt% sulfuric acid were obtained as the following order : sawdust > rice straw > copying paper > newspaper when conducting enzymatic saccharification.

High β-Glucosidase Secretion in Saccharomyces cerevisiae Improves the Efficiency of Cellulase Hydrolysis and Ethanol Production in Simultaneous Saccharification and Fermentation

Bioethanol production from lignocellulose is considered as a sustainable biofuel supply. However, the low cellulose hydrolysis efficiency limits the cellulosic ethanol production. The cellulase is strongly inhibited by the major end product cellobiose, which can be relieved by the addition of β-glucosidase. In this study, three β-glucosidases from different organisms were respectively expressed in Saccharomyces cerevisiae and the β-glucosidase from Saccharomycopsis fibuligera showed the best activity (5.2 U/ml). The recombinant strain with S. fibuligera β-glucosidase could metabolize cellobiose with a specific growth rate similar to the control strain in glucose. This recombinant strain showed higher hydrolysis efficiency in the cellulose simultaneous saccharification and fermentation, when using the Trichoderma reesei cellulase, which is short of the β-glucosidase activity. The final ethanol concentration was 110% (using Avicel) and 89% (using acid-pretreated corncob) higher than the control strain. These results demonstrated the effect of β-glucosidase secretion in the recombinant S. cerevisiae for enhancing cellulosic ethanol conversion.

Techno‐economic analysis and life‐cycle assessment of cellulosic isobutanol and comparison with cellulosic ethanol and n‐butanol

AbstractThis work presents a detailed analysis of the production design and economics of the cellulosic isobutanol conversion processes and compares cellulosic isobutanol with cellulosic ethanol and n‐butanol in the areas of fuel properties and engine compatibility, fermentation technology, product purification process design and energy consumption, overall process economics, and life cycle assessment. Techno‐economic analysis is used to understand the current stage of isobutanol process development and the impact of key parameters on the overall process economics in a consistent way (i.e. using the same financial assumptions, plant scale, and cost basis). The calculated minimum isobutanol selling price is $3.62/gasoline gallon equivalent ($/GGE) – similar to $3.66/GGE from the n‐butanol process and higher than $3.26/GGE from the cellulosic ethanol conversion process. At the conversion stage, the n‐butanol process emits the most direct CO2, at 26.42 kg CO2/GGE. Isobutanol and ethanol plants have relatively similar CO2 emissions, at 21.91 kg CO2/GGE and 21.01 kg CO2/GGE, respectively. The consumptive water use of the biorefineries increases in the following order: ethanol (8.19 gal/GGE) < isobutanol (8.98 gal/GGE) < n‐butanol (10.84 gal/GGE). Field‐to‐wheel life cycle greenhouse gas (GHG) emissions for the ethanol and n‐butanol conversion processes are similar at 4.3 and 4.5 kg CO2‐eq/GGE, respectively. The life cycle GHG emissions result for the isobutanol conversion process is 5.0 kg CO2‐eq/GGE, approximately 17% higher than that of ethanol. The life cycle fossil fuel consumption is 39 MJ/GGE for n‐butanol, 43 MJ/GGE for ethanol and 51 MJ/GGE for isobutanol. The energy return on investment for each biofuel is also determined and compared: isobutanol (2.2:1) < ethanol (2.7:1) < n‐butanol (2.8:1). © 2013 Society of Chemical Industry and John Wiley & Sons, Ltd

Challenges of cellulosic ethanol production from xylose-extracted corncob residues

Xylose-extracted corncob residue (X-ER), a byproduct from the xylose production industry, is a potential cellulose-rich energy resource. However, attempts to achieve large-scale production of cellulosic ethanol using X-ER have been unsatisfactory due to a lack of understanding of the substrate. This study presents the first characterization of the X-ER to evaluate its potential utilization in the sequential production of cellulosic ethanol. The current dilute acid treatment procedures used for the corncobs by the xylose-production industry were insufficient for efficient deconstruction of cellulose structure to release available sugars for subsequent cellulosic ethanol conversion. After a secondary dilute acid hydrolysis of the X-ER, an additional 30% hemicellulose was recovered. In addition, a more efficient enzymatic hydrolysis of X-ER was observed resulting in a significantly higher yield of glucose conversion compared with an untreated X-ER control. These results suggest X-ER can be utilized for cellulosic ethanol production. However, improved corncob pretreatment procedures are needed for economical cellulosic ethanol conversion.

Quantitative Trait Loci and Trait Correlations for Maize Stover Cell Wall Composition and Glucose Release for Cellulosic Ethanol

In cellulosic ethanol production, the efficiency of converting maize (Zea mays L.) stover into fermentable sugars partly depends on the stover cell wall structure. Breeding for improved stover quality for cellulosic ethanol may benefit from the use of molecular markers. However, limited quantitative trait loci (QTL) studies have been published for maize stover cell wall components, and no QTL study has been published for glucose release (GLCRel) from stover by a cellulosic ethanol conversion process. Our objectives were to characterize the relationships among stover cell wall components and GLCRel, and to identify QTL with major effects, if any, influencing stover cell wall composition and GLCRel. Testcrosses of 223 intermated B73 × Mo17 recombinant inbreds and the parent lines were analyzed for cell wall composition and GLCRel after acid pretreatment and enzymatic hydrolysis. As expected, glucose (GLC), xylose (XYL), and Klason lignin (KL) composed the bulk (∼72%) of the stover dry matter. Significant genetic variance and moderate heritability were observed for all traits. Genetic and phenotypic correlations among traits were generally in the favorable direction but also reflected the complexity of maize stover cell wall composition. We found 152 QTL, mostly with small effects, for GLCRel and cell wall components on both a dry matter and cell wall basis. Because no major QTL were found, methods that increase the frequency of favorable QTL alleles or that predict performance based on markers would be appropriate in marker‐assisted breeding for maize stover quality for cellulosic ethanol.

Evolutionarily engineered ethanologenic yeast detoxifies lignocellulosic biomass conversion inhibitors by reprogrammed pathways.

Lignocellulosic biomass conversion inhibitors, furfural and HMF, inhibit microbial growth and interfere with subsequent fermentation of ethanol, posing significant challenges for a sustainable cellulosic ethanol conversion industry. Numerous yeast genes were found to be associated with the inhibitor tolerance. However, limited knowledge is available about mechanisms of the tolerance and the detoxification of the biomass conversion inhibitors. Using a robust standard for absolute mRNA quantification assay and a recently developed tolerant ethanologenic yeast Saccharomyces cerevisiae NRRL Y-50049, we investigate pathway-based transcription profiles relevant to the yeast tolerance and the inhibitor detoxification. Under the synergistic inhibitory challenges by furfural and HMF, Y-50049 was able to withstand the inhibitor stress, in situ detoxify furfural and HMF, and produce ethanol, while its parental control Y-12632 failed to function till 65 h after incubation. The tolerant strain Y-50049 displayed enriched genetic background with significantly higher abundant of transcripts for at least 16 genes than a non-tolerant parental strain Y-12632. The enhanced expression of ZWF1 appeared to drive glucose metabolism in favor of pentose phosphate pathway over glycolysis at earlier steps of glucose metabolisms. Cofactor NAD(P)H generation steps were likely accelerated by enzymes encoded by ZWF1, GND1, GND2, TDH1, and ALD4. NAD(P)H-dependent aldehyde reductions including conversion of furfural and HMF, in return, provided sufficient NAD(P)(+) for NAD(P)H regeneration in the yeast detoxification pathways. Enriched genetic background and a well maintained redox balance through reprogrammed expression responses of Y-50049 were accountable for the acquired tolerance and detoxification of furfural to furan methanol and HMF to furan dimethanol. We present significant gene interactions and regulatory networks involved in NAD(P)H regenerations and functional aldehyde reductions under the inhibitor stress.

Woody biomass availability for bioethanol conversion in Mississippi

Abstract This study evaluated woody biomass from logging residues, small-diameter trees, mill residues, and urban waste as a feedstock for cellulosic ethanol conversion in Mississippi. The focus on Mississippi was to assess in-state regional variations and provide specific information of biomass estimates for those facilities interested in locating in Mississippi. Supply and cost of four woody biomass sources were derived from Forest Inventory Analysis (FIA) information, a recent forest inventory conducted by the Mississippi Institute for Forest Inventory, and primary production costs. According to our analysis, about 4.0 million dry tons of woody biomass are available for production of up to 1.2 billion liters of ethanol each year in Mississippi. The feedstock consists of 69% logging residues, 21% small-diameter trees, 7% urban waste, and 3% mill residues. Of the total, 3.1 million dry tons (930 million liters of ethanol) can be produced for $34 dry ton−1 or less. Woody biomass from small-diameter trees is more expensive than other sources of biomass. Transportation costs accounted for the majority of total production costs. A sensitivity analysis indicates that the largest impacts in production costs of ethanol come from stumpage price of woody biomass and technological efficiency. These results provide a valuable decision support tool for resource managers and industries in identifying parameters that affect resource magnitude, type, and location of woody biomass feedstocks in Mississippi.

An Economic Assessment of Traditional and Cellulosic Ethanol Technologies

Ethanol is a clean, renewable fuel that can be produced from biomass resources in nearly every region of the United States. Today in the U.S. the majority of ethanol is produced from corn, a practice that relies on subsidies to compete economically with gasoline and diverts food supplies. Significant growth of the ethanol industry will depend on the development of new processes that convert cellulosic biomass from non-food crops and waste materials into ethanol. The purpose of this research is to compare the private and total social cost of producing ethanol using conventional methods from corn to the production of ethanol from cellulosic biomass feedstocks, and to analyze the sensitivity of the technology to key variables such as capital costs, feedstock and energy prices, and estimates of social costs. Results indicate that existing ethanol technologies hold an economic advantage due primarily to the larger capital costs and greater complexity associated with the cellulosic ethanol conversion process. However, under certain economic conditions cellulosic technology is competitive with traditional methods.