Lignocellulosic Ethanol Production Research Articles

Acetic acid- and furfural-based adaptive evolution of Saccharomyces cerevisiae strains for improving stress tolerance and lignocellulosic ethanol production

Acetic acid and furfural are known as prevalent inhibitors deriving from pretreatment during lignocellulosic ethanol production. They negatively impact cell growth, glucose uptake and ethanol biosynthesis of Saccharomyces cerevisiae strains. Development of industrial S. cerevisiae strains with high tolerance towards these inhibitors is thus critical for efficient lignocellulosic ethanol production. In this study, the acetic acid or furfural tolerance of different S. cerevisiae strains could be significantly enhanced after adaptive evolution via serial cultivation for 40 generations under stress conditions. The acetic acid-based adaptive strain SPSC01-TA9 produced 30.5 g·L–1 ethanol with a yield of 0.46 g·g–1 in the presence of 9 g·L–1 acetic acid, while the acetic acid/furfural-based adaptive strain SPSC01-TAF94 produced more ethanol of 36.2 g·L–1 with increased yield up to 0. 49 g·g–1 in the presence of both 9 g·L–1 acetic acid and 4 g·L–1 furfural. Significant improvements were also observed during non-detoxified corn stover hydrolysate culture by SPSC01-TAF94, which achieved ethanol production and yield of 29.1 g·L–1 and 0.49 g·g–1, respectively, the growth and fermentation efficiency of acetic acid/furfural-based adaptive strain in hydrolysate was 95% higher than those of wildtype strains, indicating the acetic acid- and furfural-based adaptive evolution strategy could be an effective approach for improving lignocellulosic ethanol production. The adapted strains developed in this study with enhanced tolerance against acetic acid and furfural could be potentially contribute to economically feasible and sustainable lignocellulosic biorefinery.

Systematic investigation of TetR-family transcriptional regulators and their roles on lignocellulosic inhibitor acetate tolerance in Zymomonas mobilis.

TetR-family transcriptional regulators are widely distributed among bacteria and involved in various cellular processes such as multidrug and inhibitor resistance. Zymomonas mobilis is a industrial bacterium for lignocellulosic ethanol production. Although TetR-family regulators and their associated RND-family efflux pumps in Z. mobilis have been identified to be differentially expressed under various inhibitors and stressful conditions, there are no systematic investigation yet. In this study, bioinformatic analyses indicated that there are three TetR-family transcriptional regulators (ZMO0281, ZMO0963, ZMO1547) and two RND-family efflux pumps (ZMO0282-0285, ZMO0964-0966) adjacent to corresponding TetR-family regulators of ZMO0281 and ZMO0963 in Z. mobilis. Genetics studies were then carried out with various mutants of TetR-family regulators constructed, and ZMO0281 was characterized to be related to acetate tolerance. Combining transcriptomics and dual-reporter gene system, this study demonstrated that three TetR-family regulators repressed their adjacent genes specifically. Moreover, TetR-family regulator ZMO0281 might also be involved in other cellular processes in the presence of acetate. In addition, the upregulation of RND-family efflux pumps due to ZMO0281 deletion might lead to an energy imbalance and decreased cell growth in Z. mobilis under acetate stress. The systematic investigation of all three TetR-family regulators and their roles on a major lignocellulosic inhibitor acetate tolerance in Z. mobilis thus not only unravels the molecular mechanisms of TetR-family regulators and their potential cross-talks on regulating RND-family efflux pumps and other genes in Z. mobilis, but also provides guidance on understanding the roles of multiple regulators of same family in Z. mobilis and other microorganisms for efficient lignocellulosic biochemical production.

Characterization of a thermophilic and glucose-tolerant GH1 β-glucosidase from hot springs and its prospective application in corn stover degradation.

β-Glucosidase serves as the pivotal rate-limiting enzyme in the cellulose degradation process, facilitating the hydrolysis of cellobiose and cellooligosaccharides into glucose. However, the widespread application of numerous β-glucosidases is hindered by their limited thermostability and low glucose tolerance, particularly in elevated-temperature and high-glucose environments. This study presents an analysis of a β-glucosidase gene belonging to the GH1 family, denoted lqbg8, which was isolated from the metagenomic repository of Hehua hot spring located in Tengchong, China. Subsequently, the gene was cloned and heterologously expressed in Escherichia coli BL21(DE3). Post expression, the recombinant β-glucosidase (LQBG8) underwent purification through a Ni affinity chromatography column, thereby enabling the in-depth exploration of its enzymatic properties. LQBG8 had an optimal temperature of 70°C and an optimum pH of 5.6. LQBG8 retained 100 and 70% of its maximum activity after 2-h incubation periods at 65°C and 70°C, respectively. Moreover, even following exposure to pH ranges of 3.0-10.0 for 24 h, LQBG8 retained approximately 80% of its initial activity. Notably, the enzymatic prowess of LQBG8 remained substantial at glucose concentrations of up to 3 M, with a retention of over 60% relative activity. The kinetic parameters of LQBG8 were characterized using cellobiose as substrate, with Km and Vmax values of 28 ± 1.9 mg/mL and 55 ± 3.2 μmol/min/mg, respectively. Furthermore, the introduction of LQBG8 (at a concentration of 0.03 mg/mL) into a conventional cellulase reaction system led to an impressive 43.7% augmentation in glucose yield from corn stover over a 24-h period. Molecular dynamics simulations offered valuable insights into LQBG8's thermophilic nature, attributing its robust stability to reduced fluctuations, conformational changes, and heightened structural rigidity in comparison to mesophilic β-glucosidases. In summation, its thermophilic, thermostable, and glucose-tolerant attributes, render LQBG8 ripe for potential applications across diverse domains encompassing food, feed, and the production of lignocellulosic ethanol.

The impact of lignocellulosic ethanol production on the environment from a life cycle perspective

In the production process of cellulosic ethanol, all sections are closely related. It is difficult to study its producing law and key factors that affect the process only by experimental means. Computer simulation has a powerful data mining ability, which can analyze, design, and evaluate the production process of cellulosic ethanol. Specifically, it can use data calculated by Aspen Plus before, and analyze the impact of cellulose ethanol production on the environment by life cycle assessment (LCA). Results show that the total environmental impact value of the whole process is 0.20∼0.37 person·a/t. And in the whole process, the growth stage and the production stage have the biggest impact.

Molecular mechanism of engineered Zymomonas mobilis to furfural and acetic acid stress

Acetic acid and furfural (AF) are two major inhibitors of microorganisms during lignocellulosic ethanol production. In our previous study, we successfully engineered Zymomonas mobilis 532 (ZM532) strain by genome shuffling, but the molecular mechanisms of tolerance to inhibitors were still unknown. Therefore, this study investigated the responses of ZM532 and its wild-type Z. mobilis (ZM4) to AF using multi-omics approaches (transcriptomics, genomics, and label free quantitative proteomics). Based on RNA-Seq data, two differentially expressed genes, ZMO_RS02740 (up-regulated) and ZMO_RS06525 (down-regulated) were knocked out and over-expressed through CRISPR-Cas technology to investigate their roles in AF tolerance. Overall, we identified 1865 and 14 novel DEGs in ZM532 and wild-type ZM4. In contrast, 1532 proteins were identified in ZM532 and wild-type ZM4. Among these, we found 96 important genes in ZM532 involving acid resistance mechanisms and survival rates against stressors. Furthermore, our knockout results demonstrated that growth activity and glucose consumption of mutant strains ZM532∆ZMO_RS02740 and ZM4∆ZMO_RS02740 decreased with increased fermentation time from 42 to 55 h and ethanol production up to 58% in ZM532 than that in ZM532∆ZMO_RS02740. Hence, these findings suggest ZMO_RS02740 as a protective strategy for ZM ethanol production under stressful conditions.

Biomass pretreatment method affects the physicochemical properties of biochar prepared from residues of lignocellulosic ethanol production

Biomass pretreatment method affects the physicochemical properties of biochar prepared from residues of lignocellulosic ethanol production

Regulation of biofilm formation in Zymomonas mobilis to enhance stress tolerance by heterologous expression of pfs and luxS.

Zymomonas mobilis is a potential alternative of Saccharomyces cerevisiae to produce cellulosic ethanol with strengths in cofactor balance, but its lower tolerance to inhibitors in the lignocellulosic hydrolysate restricts the application. Although biofilm can improve bacteria stress tolerance, regulating biofilm formation in Z. mobilis is still a challenge. In this work, we constructed a pathway by heterologous expressing pfs and luxS from Escherichia coli in Z. mobilis to produce AI-2 (autoinducer 2), a universal quorum-sensing signal molecule, to control cell morphology for enhancing stress tolerance. Unexpectedly, the results suggested that neither endogenous AI-2 nor exogenous AI-2 promoted biofilm formation, while heterologous expression of pfs can significantly raise biofilm. Therefore, we proposed that the main factor in assisting biofilm formation was the product accumulated due to heterologous expression of pfs, like methylated DNA. Consequently, ZM4::pfs produced more biofilm, which presented an enhanced tolerance to acetic acid. All these findings provide a novel strategy to improve the stress tolerance of Z. mobilis by enhancing biofilm formation for efficient production of lignocellulosic ethanol and other value-added chemical products.

Physiological and transcriptome analyses of Kluyveromyces marxianus reveal adaptive traits in stress response.

Kluyveromyces marxianus is a non-conventional yeast with outstanding physiological characteristics and a high potential for lignocellulosic ethanol production. However, achieving high ethanol productivity requires overcoming several biotechnological challenges due to the cellular inhibition caused by the inhibitors present in the medium. In this work, K. marxianus SLP1 was adapted to increase its tolerance to a mix of inhibitory compounds using the adaptive laboratory evolution strategy to study the adaptation and stress response mechanisms used by this non-Saccharomyces yeast. The fermentative and physiological parameters demonstrated that the adapted K. marxianus P8 had a better response against the synergistic effects of multiple inhibitors because it reduced the lag phase from 12 to 4 h, increasing the biomass by 40% and improving the volumetric ethanol productivity 16-fold than the parental K. marxianus SLP1. To reveal the effect of adaptation process in P8, transcriptome analysis was carried out; the result showed that the basal gene expression in P8 changed, suggesting the biological capability of K. marxianus to activate the adaptative prediction mechanism. Similarly, we carried out physiologic and transcriptome analyses to reveal the mechanisms involved in the stress response triggered by furfural, the most potent inhibitor in K. marxianus. Stress response studies demonstrated that P8 had a better physiologic response than SLP1, since key genes related to furfural transformation (ALD4 and ALD6) and stress response (STL1) were upregulated. Our study demonstrates the rapid adaptability of K. marxianus to stressful environments, making this yeast a promising candidate to produce lignocellulosic ethanol. KEY POINTS: • K. marxianus was adapted to increase its tolerance to a mix of inhibitory compounds • The basal gene expression of K. marxianus changed after the adaptation process • Adapted K. marxianus showed a better physiological response to stress by inhibitors • Transcriptome analyses revealed key genes involved in the stress response.

The Relationship between Structural Features of Lignocellulosic Materials and Ethanol Production Yield

Lignocellulosic materials are a mixture of natural polymers which can be considered a great alternative source of chemical products and energy. Hence, pinewood, poplar wood, and rice straw, as representatives of different types of lignocelluloses, were subjected to several pretreatment types in order to increase ethanol production yield. All pretreatments increased enzymatic hydrolysis and ethanol yield, specifically pretreatment with phosphoric acid. This pretreatment increased ethanol yields by 304.6% and 273.61% for poplar wood and pinewood, respectively, compared to untreated substrates. In addition, a number of analyses, including a BET test, buffering capacity, crystallinity, accessible surface area, and composition measurement, were conducted on the pretreated substrates to investigate their structural modifications in detail. Accessible surface area, as one of the most important parameters for performance of enzymes and microorganisms in the fermentation process, was examined by the water retention value test. The results of this method (using centrifuge) showed that the maximum accessible surface area was related to the pretreated samples with phosphoric acid so that it increased WRV to 132.19%, 149.41%, and 68.44% for poplar wood, pinewood, and rice straw, respectively, as compared to untreated substrates. On the whole, pretreatments restructured and opened up the tangled structure of lignocelluloses, resulting in a considerable increase in ethanol yields. Moreover, in this study, for the first time, a new correlation was presented for each substrate which indicates the relationship between ethanol yield and structural features of the lignocellulosic substrate.

The crop residue conundrum: Maintaining long-term soil organic carbon stocks while reinforcing the bioeconomy, compatible endeavors?

Crop residues are a key bulk feedstock for supplying renewable carbon for bioenergy production and the broader bioeconomy without compromising food security. However, it is frequently advised to harvest no more than half of this potential to ensure the preservation of soil organic carbon (SOC) stocks. In this study, we challenge this recommendation and demonstrate that the crop residue potential allowing to maintain long-term SOC stocks is spatially differentiated and strongly dependent upon the bioeconomy conversion pathway for which it is intended. We assessed the interaction between the residues' usage for the bioeconomy and the maintenance of SOC stocks over 100 years by considering the coproduct return to soils from five bioeconomy pathways: pyrolysis, gasification, hydrothermal liquefaction, anaerobic digestion, and lignocellulosic ethanol production. To compare the long-term SOC changes from these scenarios against a reference where crop residues are unharvested, we developed a novel framework, applicable to any site or region, by coupling a SOC model that includes recalcitrant organic matter deriving from a bioeconomy calculation module. The adapted SOC model considers the recalcitrance to degradation of each coproduct, while the bioeconomy module determines the share of carbon from the crop residues ending in the coproducts. The framework was tested and applied with a high spatial resolution (>60,000 simulation units) to the context of French croplands over the period of 2020–2120, with state-of-the-art sensitivity analyses. The case study results revealed, among others, that an additional crop residue potential equivalent to 71–225 PJ (pathway-dependent) could be available for the French bioeconomy without SOC decreases, compared to applying a stringent removal limit of 31.5%.

Integrated Method of Pretreatment, Saccharification and Electro-Fermentation of Saccharum Spontaneum to Produce Bioethanol Using Microbial Consortium

Production of lignocellulosic ethanol using microbial consortium can be an economical strategy. Consortium of ligninolytic and cellulolytic microbe(s) that can depolymerize lignin and hydrolyze cellulose along with fermenting yeast (CDBT-2) can replace the separate techniques used for biomass pretreatment and hydrolysis prior to fermentation. So, the aim of this study was to screen fungi capable of producing laccase (lignin depolymerizing enzyme) and cellulase (converts cellulose to fermentable sugars), and use the strains along with Saccharomyces cerevisiae (CDBT-2) to develop an integrated method of simultaneous pretreatment saccharification and electro-fermentation of Saccharum spontaneum powder to produce ethanol. 18-s rRNA sequencing and BLAST analysis of screened isolates confirmed, cellulolytic isolate (F1) resembled as Aspergillus niger and, ligninocellulolytic isolate (F2) resembled to Ganoderma sessile. These fungi were used along with CDBT2 for ethanol production. Different consortiums sets of these isolates were tested for fermentation efficiency. All the analysis were done using crude untreated as well as hot water pretreated S. spontaneum powder to compare the ethanol production efficiency. In controlled environment of 28oC, shaking at 80 rpm and 5.0 pH, culture with consortium of G. sessile and CDBT2 produced higher ethanol with yield 57 mg/g biomass powder followed by consortium of G. sessile, A. niger and CDBT2 with yield of 44±0.18 mg/g biomass. Monoculture of G. sessile produced higher amount of phenol ie 292.48±17.74 µg/mL. Similarly, G. sessile, A. niger and CDBT2 consortium produced higher furfural of 58.48±3.86 µg/mL. However, the amount of phenol and furfural generated were very low in both cases. The ethanol yield was higher in crude biomass compared to hot water pretreated S. spontaneum powder. In addition, electro-fermentation of crude S. spontaneum enhanced ethanol production by 12% with G. sessile and CDBT2 consortium while 18% with the set of G. sessile, A. niger and CDBT2 consortium. This monophasic method of integrated pretreatment saccharification and electro-fermentation can be an alternative approach for lignocellulosic bioethanol production as it does not require separate pretreatment and fermentation approach.Keywords: Bioethanol, Saccharum spontaneum, microbial consortium, simultaneous saccharification and fermentation, and electro-fermentation Figure 1

Environmental assessment of the valorization of glycerol for the production of hyperthermophilic β-glucosidase under a biorefinery approach

Bioethanol production technologies from lignocellulosic biomass are not yet optimized and do not compete economically with first-generation bioethanol production. Strategies have been investigated to produce more active, stable and temperature-tolerant enzymes to be used for biomass hydrolysis such as the hyperthermophilic β-glucosidase produced by Yarrowia lipolytica. The use of this strain offers an additional competitive advantage, as it can use glycerol stream from the biodiesel process as a carbon source. In this way, not only is a by-product of biofuel production used, but the enzyme could be applied in the production of lignocellulosic ethanol, increasing the value chain by closing the bioeconomy cycle. To this end, large-scale process modelling of β-glucosidase production has been developed to collect the inventory data needed for life cycle assessment methodology. The fermentation stage is the largest contributor to environmental impacts, with electricity being the main hotspot identified, contributing more than 50% in most impact categories. Residual glycerol has also been identified as a critical input, with a significant contribution in some categories. To improve the environmental profile, a sensitivity analysis has been carried out considering reductions in electricity and heat consumption, and other alternative oil-based resources for the production of biodiesel. This analysis identified that large environmental reductions could be achieved, which makes the valorization of the glycerol obtained as a side stream of biodiesel production more realistic.

Production of bioethanol from Saccharum spontaneum by simultaneous saccharification and electro-fermentation using mixed culture of microbes

Fungal consortium of a cellulolytic isolate F1 (Aspergillus niger) and/or ligninocellulolytic isolate F2 (Ganoderma sessile) along with Saccharomyces cerevisiae (CDBT2) were used for integrated approach of saccharification, fermentation and electrofermentation of Saccharum spontaneum biomass to produce ethanol. Different consortium sets of these isolates were used to observe fermentation efficiency. All the analysis were done using crude as well as hot water pretreated S. spontaneum powder to compare the ethanol production efficiency. The ethanol yield was found to be higher in crude biomass compared to hot water pretreated S. spontaneum biomass. At 28°C, 80 rpm and 5.0 pH, simultaneous culture with combination of G. sessile and CDBT2 produced higher amount of ethanol (57 mg/g crude biomass) followed by combination of G. sessile, A. niger and CDBT2 (44 ± 0.18 mg/g crude biomass). However, the amount of phenol and furfural generated were very low in both cases. In addition, electro-fermentation enhanced ethanol production by 12% and 18% with a combination culture of G. sessile, CDBT2 and G. sessile, A. niger, CDBT2 respectively. This method of lignocellulosic ethanol production by simultaneous saccharification and electrofermentation can be developed as an alternative approach for cost effective production of bioethanol.

Using organosolv pretreatment with acid wastewater for enhanced fermentable sugar and ethanol production from rubberwood waste

The intensive utilization of water in the pretreatment of substrates for lignocellulosic ethanol production negatively impacts its economic feasibility. This study aimed to develop a method of pretreatment using acid wastewater to enhance the conversion of rubberwood waste into fermentable sugar and ethanol. The water was replaced with acid wastewater in hydrothermal pretreatments with and without alkaline post-treatment, and organosolv pretreatment (wastewater:ethanol 50:50). The organosolv pretreatment at 190 °C maximized the fermentable sugar concentration (43.60 g/L) and yield (301.0 ± 13.3 kg/ton rubberwood waste) because of partial delignification and physical redistribution of the rubberwood waste matrix. It also demonstrated markedly better reduction of water utilization, wastewater generation, and energy consumption than other pretreatments. In addition, the hydrolysate derived from the organosolv pretreatment was chosen for ethanol production. Saccharomyces cerevisiae showed better ethanol conversion rate than Kluyveromyces marxianus when the hydrolysate predominantly contained glucose, resulting in an ethanol production of 0.14 g/g rubberwood waste. The pretreatment and fermentation were effective and exhibited the potential to guide lignocellulosic ethanol production in collaboration with latex factories (acid wastewater sources) in future practical applications.

Evaluation of lignocellulose degradation and ethanol production via dilute acid and alkali pretreatment of hybrid Pennisetum

Chemically pretreating lignocellulose has been widely used in the production of clean energy. An energy grass called hybrid Pennisetum was pretreated with different H2SO4 and NaOH concentrations to analyse the effects of an acid and alkali pretreatment on the degradation of hybrid Pennisetum lignocellulose. The chemical composition, hydrolyzed sugar yield, and microstructure were determined. The results showed that a NaOH pretreatment retained more cellulose and removed more lignins, while a H2SO4 pretreatment significantly inhibited the removal of lignins and increased hemicellulose degradation. X-ray diffraction analysis indicated that increasing the concentration of NaOH or H2SO4 could intensify the cellulose polymerization; this effect was stronger with NaOH compared to H2SO4. Scanning electron microscopy images showed that the cellulose bundles pretreated with H2SO4 were arranged tightly, while the cellulose bundles pretreated with NaOH were loose. Fourier transform infrared spectroscopy exhibited that the ether/ester bond between lignins and carbohydrates was broken by the H2SO4 pretreatment. The NaOH pretreatment increased the number of carbonyl and hydroxyl groups, resulting in more hydrophilic lignins, enabling greater facile degradation. Although the two methods are different in terms of destroying the structure of lignocellulose, the NaOH pretreatment can retain more cellulose, which is beneficial to subsequent ethanol production.

Crops with potential for diclosulam remediation and concomitant bioenergy production

The objective of this work was to evaluate crops for their ability to phytoremediate diclosulam residues in the soil and produce lignocellulosic ethanol. Physiological characteristics, biomass production, soil cover rate, fermentable sugar production and lignocellulosic ethanol production potential of the crops were evaluated in soil with diclosulam residues. The experimental design was a randomized block with four replications. The treatments were arranged in a 4 × 2 factorial scheme with the following crops as the first factor: Avena sativa, Canavalia ensiformis, Mucuna aterrima, and Pennisetum glaucum. The second factor was the presence or absence of the herbicide diclosulam in the soil (30 g ha−1). The physiological variables of the plant species were not affected by the presence of diclosulam; the soil cover of P. glaucum was lower in the area with diclosulam, with a value of 26%. The levels of glucose were not affected by the presence of diclosulam in A. sativa, C. ensiformis, and M. aterrima, indicate not change the estimated yield of ethanol for this species. Avena sativa and Pennisetum glaucum have the potential to phytoremediate soils containing diclosulam residues, with concomitant lignocellulosic ethanol production ability.

Formate Dehydrogenase Improves the Resistance to Formic Acid and Acetic Acid Simultaneously in Saccharomyces cerevisiae.

Bioethanol from lignocellulosic biomass is a promising and sustainable strategy to meet the energy demand and to be carbon neutral. Nevertheless, the damage of lignocellulose-derived inhibitors to microorganisms is still the main bottleneck. Developing robust strains is critical for lignocellulosic ethanol production. An evolved strain with a stronger tolerance to formate and acetate was obtained after adaptive laboratory evolution (ALE) in the formate. Transcriptional analysis was conducted to reveal the possible resistance mechanisms to weak acids, and fdh coding for formate dehydrogenase was selected as the target to verify whether it was related to resistance enhancement in Saccharomyces cerevisiae F3. Engineered S. cerevisiae FA with fdh overexpression exhibited boosted tolerance to both formate and acetate, but the resistance mechanism to formate and acetate was different. When formate exists, it breaks down by formate dehydrogenase into carbon dioxide (CO2) to relieve its inhibition. When there was acetate without formate, FDH1 converted CO2 from glucose fermentation to formate and ATP and enhanced cell viability. Together, fdh overexpression alone can improve the tolerance to both formate and acetate with a higher cell viability and ATP, which provides a novel strategy for robustness strain construction to produce lignocellulosic ethanol.

Saccharomyces cerevisiae and newly isolated Candida boidinii co-fermentation of industrial tea waste for improved bioethanol production

ABSTRACT Although bioethanol is a suitable alternative to fossil-based fuels, limited technologies or lack of microorganisms with a broad substrate spectrum are serious bottlenecks for lignocellulosic ethanol production. In this context, the current study aimed to optimize bioethanol production from industrial tea waste by applying co-fermentation strategy for the first time. For this purpose, newly isolated xylose fermenter Candida boidinii and Saccharomyces cerevisiae produced 12.1 g/L and 14.1 g/L bioethanol, respectively, when the yeasts were cultivated alone in 20% (w/v) dilute acid pretreated and enzymatically hydrolyzed industrial tea waste. Bioethanol production increased to 21.9 g/L, and 70% improvement was achieved when the co-fermentation strategy was applied under the same conditions. This study shows that co-fermentation of industrial tea waste is an economically viable choice for value-added bioethanol production.

Coupling hydrothermal liquefaction and aqueous phase reforming for integrated production of biocrude and renewable H2

AbstractLignin‐rich stream from lignocellulosic ethanol production was converted into biocrude by continuous hydrothermal liquefaction (HTL) while hydrogen was produced by aqueous phase reforming (APR) of the HTL aqueous by‐product. The effects of Na2CO3 and NaOH were investigated both in terms of processability of the feedstock as well as yield and composition of the obtained products. A maximum biocrude yield of 27 wt% was reached in the NaOH‐catalyzed runs. A relevant amount of dissolved phenolics were detected in the co‐produced aqueous phase (AP), and removed by liquid–liquid extraction using butyl acetate or diethyl ether, preserving the APR catalyst stability and reaching an hydrogen yield up to 146 mmol H2 L−1 AP. Preliminary mass balances integrating HTL and APR showed that the hydrogen provided by APR may account for up to 46% of the hydrogen amount theoretically required for upgrading the HTL biocrude, thus significantly improving the process performance and sustainability.

Techno-Economic Evaluation of Novel Hybrid Biomass and Electricity-Based Ethanol Fuel Production

In order to limit climate change, fast greenhouse gas reductions are required already before 2030. Ethanol commonly produced by fermentation of sugars derived either from starch-based raw material such as corn, or lignocellulosic biomass is an established fuel decarbonizing the transport sector. We present a novel selective and flexible process concept for the production of ethanol with electricity and lignocellulosic biomass as main inputs. The process consists of several consecutive steps. First synthesis gas from gasification of biomass is purified by filtration and reforming and fed to methanol synthesis. The produced methanol is fed to acetic acid synthesis, together with a carbon monoxide-rich stream separated from the synthesis gas by membranes. Finally, acetic acid is hydrogenated to yield ethanol. With the exception of acetic acid hydrogenation, the overall process consists of technically mature subprocesses. Each process step was modelled in Aspen Plus to generate the mass and energy balances for the overall process. Additionally, the CO2 emissions and economic feasibility were assessed. Three separate cases were investigated. In the first two cases, the syngas carbon (CO and CO2) was split between methanol and acetic acid synthesis. The cases included either allothermal (case A) or electrically heated reforming (case B). In case C, maximum amount of CO was sent to acetic acid synthesis to maximize the acetic acid output, requiring a small additional carbon dioxide input to methanol synthesis. In all cases, additional hydrogen to methanol synthesis was provided by water electrolysis. Each case was designed at biomass input of 27.9 MW and the electrolyzer electricity requirement between 36 and 43.5 MW, depending on the case. The overall energy efficiency was calculated at 53–57%, and carbon efficiencies were above 90%. The lowest levelized cost of ethanol was 0.65 €/l, at biomass cost of 20 €/MWh and electricity cost of 45 €/MWh and production scale of approximately 42 kt ethanol per year. The levelized cost is competitive with the current biological route for lignocellulosic ethanol production. The ethanol price is very sensitive to the electricity cost, varying from 0.56 to 0.74 €/l at ±30% variation in electricity cost.