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Corn fiber, a by-product from the corn-processing industry, is an attractive feedstock for cellulosic ethanol because of its rich carbohydrate content (mainly residual starch, cellulose, and hemicellulose), abundant reserves, easy collection, and almost no transportation cost. However, the complex structure and components of corn fiber, especially hemicellulose, make it difficult to be effectively hydrolyzed into fermentable sugars through enzymatic hydrolysis. This study developed a simple and easy industrialized process without detoxification treatment for high-yield ethanol produced from corn fiber. Corn fiber was pretreated by dilute acid under the conditions optimized by Box-Behnken design (0.5% H2SO4 at 105°C for 43 min), and 81.8% of total sugars, including glucose, xylose, and arabinose, could be recovered, then the mixture (solid and hydrolysates) was directly used for semi-simultaneous saccharification and fermentation without detoxification, and ethanol yield reached about 81% of the theoretical yield. [ABSTRACT FROM AUTHOR]

The use of a method of an effective delignification of lignocellulosic biomass is a key stage of designing processes of its microbiological conversion e.g. for the purposes of the production of cellulosic ethanol. The study was aimed at evaluating the effectiveness of microwave-assisted hydrotropic pretreatment using sodium cumene sulfonate (NaCS) for the delignification of pine and beech chips and wheat straw. Research results presenting the impact of process parameters of microwave-assisted hydrotropic delignification confirm a high effectiveness of this method of pretreatment of lignocellulosic biomass. The observed effects included changes in the composition of the biomass and an increased susceptibility of cellulose to the subsequent enzymatic hydrolysis. The use of microwave heating combined with an addition of hydrotrope of 40% w/v NaCS and 117 PSI for 60 min enabled a reduction of the absolute concentration of lignins by 36.58% in pine chips, by 57.68% in beech chips, and by 74.08% in wheat straw. After enzymatic hydrolysis was conducted, the highest concentration of glucose: 463.27 ± 11.25 mg glucose/g (hydrolysis yield 46.76 ± 1.14%) was obtained from the wheat straw, while 327.70 ± 22.15 mg glucose/g (hydrolysis yield 35.13 ± 2.37%) was acquired from the beech chips, and only 50.77 ± 0.75 mg glucose/g (hydrolysis yield 6.63 ± 0.10%) was obtained from the pine chips. Microwave-assisted hydrotropic delignification in the optimum process conditions additionally allows a complete removal of hemicellulose from biomass, which improves the effectiveness of enzymatic hydrolysis. Due to a significant reduction of lignin and hemicellulose concentration in biomass, cellulose—which is susceptible to enzymatic hydrolysis and a source of carbon in biosynthesis processes—becomes the main biomass component. [ABSTRACT FROM AUTHOR]

Reducing the cost of cellulosic ethanol production, especially for cellulose hydrolytic enzymes, is vital to growing a sustainable and efficient cellulosic ethanol industry and bio-based economy. Using an ethanologenic yeast able to produce hydrolytic enzymes, such as Clavispora NRRL Y-50464, is one solution. NRRL Y-50464 is fast-growing and robust, and tolerates inhibitory compounds 2-furaldehyde (furfural) and 5-hydroxymethyl-2-furaldehyde (HMF) associated with lignocellulose-to-fuel conversion. It produces three forms of β-glucosidase isozymes, BGL1, BGL2, and BGL3, and ferment cellobiose as the sole carbon source. These β-glucosidases exhibited desirable enzyme kinetic parameters and high levels of enzyme-specific activity toward cellobiose and many oligosaccharide substrates. They tolerate the product inhibition of glucose and ethanol, and are stable to temperature and pH conditions. These characteristics are desirable for more efficient cellulosic ethanol production by simultaneous saccharification and fermentation. NRRL Y-50464 provided the highest cellulosic ethanol titers and conversion rates at lower cellulase loadings, using either pure cellulose or agricultural residues, as so far reported in the literature. This review summarizes NRRL Y-50464 performance on cellulosic ethanol production from refined cellulose, rice straw, and corn stover processed in various ways, in the presence or absence of furfural and HMF. This dual functional yeast has potential to serve as a prototype for the development of next-generation biocatalysts. Perspectives on continued strain development and process engineering improvements for more efficient cellulosic ethanol production from lignocellulosic materials are also discussed. [ABSTRACT FROM AUTHOR]

The addition of poly(ethylene glycol) (PEG) in the pretreatment and enzymatic hydrolysis steps has become attractive to increase second-generation ethanol titers. Recent studies have shown the benefits of using high-concentration PEG on the yeast vitality and enzymatic digestibility of untreated biomass. Thus, this study investigated the ideal concentration of PEG in the second-generation ethanol production and the best step to add it. In the acid pretreatment, PEG 4000 favored the delignification (59.2%) and reduced the non-productive adsorption of cellulases onto corn cob, but its use was not significant on the sodium carbonate–based pretreatment. Changes in PEG dosage and PEG molecular weight strongly affected the performance of enzymatic hydrolysis and simultaneous saccharification and fermentation (SSF). PEG 4000 (150 g/L) increased the production of reducing sugars by up to 30.1% and the ethanol production via SSF by up to 44.7% from untreated corn cob. In turn, PEG slightly affected the enzymatic digestibility and fermentability of the pretreated corn cob. One-pot ethanol production from corn cob was performed using PEG as an additive. The addition of 150 g/L PEG 1500 and PEG 4000 increased the ethanol yield values (> 52%) in the one-pot processes; however, the ethanol concentration was lower than in the experiments with the pretreated corn cob. SSF experiments with high solid loadings were tested using PEG 1500. Under these conditions, PEG 1500 slightly increased ethanol production, from 53.4 to 55.7 g/L ethanol (equivalent to 92.8% ethanol yield) using 20% (w/v) acid-pretreated corn cob. [ABSTRACT FROM AUTHOR]

Sugarcane bagasse (SCB), a major waste of sugar industry, provides a huge potential for cellulosic ethanol production. Efficient pretreatment method is essential to promote its bioconversion. In this study, a novel and cost-effective pretreatment "Densifying Lignocellulosic biomass with Chemicals followed by Autoclave (DLCA)" was applied on SCB for the first time. Sulfuric acid (sa) was used as the reagent. The effects of pretreatment conditions (temperature, time, acid dosage, solid biomass loading) were investigated. High solid-loading enzymatic hydrolysis and fermentations were conducted. The results showed that DLCA(sa)-SCB owned high enzymatic digestibility and high fermentability. Simultaneous saccharification and co-fermentation (SSCF) was also investigated on DLCA(sa)-SCB. A high ethanol titer of 77.51 g/L (nearly 10%, v/v) and a high ethanol yield of 234.09 g per kg SCB were obtained at 30% solid loading by SSCF with fed-batching biomass and enzymes. This is the highest ethanol titer reported on SCB without washing or detoxification. [ABSTRACT FROM AUTHOR]

In this study, the effect of residual lignin and its structural modification of pretreated sugarcane bagasse (SCB) on enzymatic hydrolysis and fermentation was comprehensively evaluated by alkali and alkaline hydrogen peroxide (AHP) pretreatment. It was found that the AHP pretreatment had a negative influence on lignin dissolution compared with alkali pretreatment accompanied by a relatively low NaOH concentration (≤1%, w/v). However, there was less significance of enzymatic hydrolysis efficiency between NaOH and AHP pretreatment with >1% NaOH loading due to a high lignin removal above 78%. In addition, PEG 6000-assisted pretreatment was also performed to stabilize the enzymatic activity and further promote enzymatic digestibility and fermentability. The highest ethanol concentration of 59.96 g/L (61.22%) was obtained with PEG 6000-assisted 1% NaOH pretreated SCB at 30% solid loading. The structural features of the residual lignin indicated that most of the side chain structures were oxidized and that some aryl-ether bonds were dissociated after the pretreatments, especially in alkali pretreatment. The reduction of molecular weight and the breaking of lignin substructures help to achieve a more effective yield of enzymatic hydrolysis and fermentation. • High glucose yield was obtained from Alkali and AHP pretreatment on sugarcane bagasse. • Surfactants assisted pretreatment has potential to shorten reaction time and accelerate cellulosic ethanol production. • High ethanol concentration (59.96 g/L) was achieved to lower the distillation cost. • The reduced Mw values and broken lignin substructures achieve more glucose yield. [ABSTRACT FROM AUTHOR]

Cellulosic ethanol has been regarded as excellent additive into petrol fuels for reduced net carbon release, and yeast fermentation is thus a crucial step for bioethanol production. In this study, three (XYL1 / Candida tropicalis, XYL2 / Candida tropicalis, XKS1 / Saccharomyces cerevisiae) genes were isolated to construct four novel vectors using gene fusion and tandem technology. Four constructs were then transformed into common Saccharomyces cerevisiae strain, leading to varied and limited xylose utilization. While two representative constructs were transformed into industrial yeast strain (SF7), the engineered SF7-Ft3 strain could consume 95% of total xylose for ethanol yield at 2.08 g/L, whereas the control strain only utilized 13% xylose with ethanol yield at 0.56 g/L. Additional XYL2 overexpression into the SF7-Ft3 strain led to consistently enhanced xylose utilization by from diverse enzymatic hydrolats of steam-exploded lignocellulose residues in three major bioenergy crops (wheat, maize, Miscanthus). These consequently increased bioethanol yields (% dry matter) and concentrations (g/L) by 11%–42%. Therefore, this study has demonstrated an applicable yeast-engineering approach for efficient xylose consumption and also provided a powerful strategy for enhancing bioethanol production in bioenergy crops. [Display omitted] • Diverse integration of XYL1, XYL2 and XKS1 for varied xylose utilizations in yeast. • Engineered industrial strain (SF7-Ft3) consumed 95% xylose for yeast fermentation. • XYL2 double-overexpression with SF7-Ft3 for further enhanced xylose consumption. • Ethanol yield and concentration were raised at 11%–42% by SF7-Ft3-X2 strain. • Integrating engineered yeast strain with steam-exploded biomass for bioethanol. [ABSTRACT FROM AUTHOR]

The finite nature of fossil fuels and the environmental impact of its use have raised interest in alternate renewable energy sources. Specifically, nonfood carbohydrates, such as lignocellulosic biomass, can be used to produce next generation biofuels, including cellulosic ethanol and other nonethanol fuels like butanol. However, currently there is no native microorganism that can ferment all lignocellulosic sugars to fuel molecules. Thus, research is focused on engineering improved microbial biocatalysts for production of liquid fuels at high productivity, titer, and yield. A clear understanding and application of the basic principles of microbial physiology and biochemistry are crucial to achieve this goal. In this review, we present and discuss the construction of microbial biocatalysts that integrate these principles with ethanol-producing Escherichia coli as an example of metabolic engineering. These principles also apply to fermentation of lignocellulosic sugars to other chemicals that are currently produced from petroleum. [ABSTRACT FROM AUTHOR]

Corn stalks have been traditionally disposed of by in-situ incineration or for the production of animal feed, due to the main crop residues in northeastern China. However, the conventional treatments of corn stalks have posed a great threat to the environment and resource utilization in recent years. Alternatively, biomass materials (corn stalks) can be widely expected to replace wood preparation in paper production. Different pretreatments can be utilized to improve the quality of pulp made from corn stover. The common-used pulping pretreatment can include various physical, chemical, and biological reactions. This study aims to explore the possibility and feasibility of a co-culture procedure of anaerobic microorganisms and yeast to pretreat the corn stalks. The culture medium was firstly prepared in the experiment, and then the resulting goat rumen fluid was filtered and centrifuged, where the centrifuged supernatant was used for the identification. After that, the supernatant of rumen fluid (anaerobic fungi: Pecoramyces sp.) and Saccharomyces cerevisiae S1145 were added to the medium for the simple co-culture. The components of the naturally air-dried corn stalks were separated, where the corn stalk husks were collected, crushed, and sieved, and the shredded husks (as a substrate) were added to the medium for the simple co-cultivation at 39°C for 72 hours. The results show that the Saccharomyces cerevisiae presented the ethanol in the metabolites of fermentation broth under the action of rumen fungi, indicating that the rumen fungi and yeast were co-cultured in vitro. The content of Saccharomyces cerevisiae was greatly contributed to the metabolism. The ethanol content in the product reached the maximum when adding 5ml of Saccharomyces cerevisiae, accounting for 32.09% of the total metabolite content. The ethanol content was increased by 23.04 percentage points with Saccharomyces cerevisiae, compared with the control without. The co-culture process of anaerobic fungi and Saccharomyces cerevisiae was utilized to pretreat the corn stalk bark, where the Saccharomyces cerevisiae was improved the ethanol production. The enzyme preparation was normally used to enzymatically hydrolyze plant cellulose in the conventional second-generation cellulosic ethanol production. Nevertheless, the enzyme preparation cannot be reused during processing, leading to the high production cost of cellulosic ethanol. Taking the rumen of ruminants as a bionic object, the living environment of anaerobic microorganisms was simulated in vitro to create the conditions for the sustainable production of cellulosic ethanol. Consequently, this experiment provided a new way to treat the corn stalks. The corn stalks after biological treatment can be used to produce the biomass pulp. The experiments have verified that the rumen anaerobic microorganisms and Saccharomyces cerevisiae can self-reproduce the corn in an artificial biomimetic environment, indicating the feasibility of straw to produce ethanol. The treatment and reuse of corn stalks can greatly contribute to the economic benefits of corn stalk pulping, while reducing the overall cost consumption for the high resource utilization. [ABSTRACT FROM AUTHOR]

Miscanthus × 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. Core ideas: Miscanthus × giganteus is suitable for cultivation in northern China.Agronomic traits were significantly correlated with biomass yield.Biomass yield of M. × giganteus increased over a period of three years.Biomass quality exhibited better suitability for ethanol production.High calorific value may be used for direct combustion and co‐combustion. [ABSTRACT FROM AUTHOR]

Anaerobes harbor some of the most efficient biological machinery for cellulose degradation, especially thermophilic bacteria, such as Acetivibrio thermocellus and Thermoclostridium stercorarium, which play a fundamental role in transferring lignocellulose into ethanol through consolidated bioprocessing (CBP). In this study, we compared activities of two cellulase systems under varying kinds of hemicellulose and cellulose. A. thermocellus was identified to contribute specifically to cellulose hydrolysis, whereas T. stercorarium contributes to hemicellulose hydrolysis. The two systems were assayed in various combinations to assess their synergistic effects using cellulose and corn stover as the substrates. Their maximum synergy degrees on cellulose and corn stover were, respectively, 1.26 and 1.87 at the ratio of 3:2. Furthermore, co-culture of these anaerobes on the mixture of cellulose and xylan increased ethanol concentration from 21.0 to 40.4 mM with a high cellulose/xylan-to-ethanol conversion rate of up to 20.7%, while the conversion rates of T. stercorarium and A. thermocellus monocultures were 19.3% and 15.2%. The reason is that A. thermocellus had the ability to rapidly degrade cellulose while T. stercorarium co-utilized both pentose and hexose, the metabolites of cellulose degradation, to produce ethanol. The synergistic effect of cellulase systems and metabolic pathways in A. thermocellus and T. stercorarium provides a novel strategy for the design, selection, and optimization of ethanol production from cellulosic biomass through CBP. [ABSTRACT FROM AUTHOR]

In a bioeconomy, the valorization of lignin beyond its use to generate energy in renewable biomass-based industries is highly attractive and economically critical. However, most of its proposed applications are highly challenging and lignin remains a highly under-utilized renewable resource despite being extensively investigated. In this study, a disc ultra-refining process was used to produce lignin nanoparticles (LNPs). The resulting spherical LNPs were investigated using spectroscopic analysis (UV, FT-IR, and 1H NMR), which revealed that no significant changes occurred in the chemical structure when compared to unprocessed lignin with the exception that a greater number of chemical groups were exposed during the process. LNPs were produced in ∼100% yield with enhanced properties, including antibacterial activity, uniform particle size, and good UV absorption. They also showed very high pH stability (pH 2–14) over several months and formed a homogenous dispersion in a polymeric matrix. In addition, the proposed method also appears to be feedstock agnostic because it was efficient in producing LNPs from lignins obtained from cellulosic ethanol and the Kraft pulping process. This study demonstrates a single-step rapid mechanical method that required no chemical additives or heating, has low-energy consumption, and which makes use of a disc ultra-refining process, as an innovative green approach to produce pH-stable LNPs in high yield with high uniformity that are highly effective as a UV protector and antibacterial agent. [ABSTRACT FROM AUTHOR]

BACKGROUND Cotton stalk (CS) was deconstructed by sequential dilute acid‐alkali pretreatment (DALP) using sulfuric acid (1%, v/v) and sodium hydroxide (3%, w/v). The enzymatic hydrolysis of DALP‐CS by cellulase was optimized by Taguchi method, employing factors like solid loading, enzyme dose, tween 80, and pH as main process variables. For bioethanol production, pretreated CS was subjected to simultaneous saccharification and fermentation (SSF) using a newly identified yeast strain Kluyveromyces marxianus JKSG‐6. RESULTS: Sequential pretreatment of CS resulted in 66% and 42% removal of hemicellulose and lignin, respectively. The same was evident from the changes in the fourier transforming infrared (FT‐IR) spectroscopy absorption spectrum. Under optimized conditions, the enzymatic hydrolysis of DALP‐CS released 375.50 ± 0.04 mg g−1 of glucose. Further, by employing K. marxianus JKSG‐6 under SSF at 42 °C, the strain produced an ethanol titer of 26.10 ± 0.05 g L−1 from pretreated CS. CONCLUSION: Sequential dilute acid‐alkali pretreatment rendered CS, a highly recalcitrant LCB feedstock for bioethanol, more accessible for hydrolysis by cellulase. SSF of the effectively pretreated CS was successfully carried out for bioethanol production by employing a newly identified yeast K. marxianus JKSG‐6. © 2021 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

Agricultural residues (e.g. corn stover (CS)) representing a huge lignocellulosic biomass waste are regarded as a promising renewable resource that can be converted to fuels and chemicals via biochemical route. Nevertheless, the unfavorable properties, such as low bulk density, contamination by microorganisms, fluffy and thereby difficult to handle, cause huge problems for biomass logistics and biomass conversion. Furthermore, traditional biomass pretreatment methods often use severe conditions, consume much energy, difficult to scale up and generate a feedstock with many toxic degradation products that inhibit fermentation. In this study, we developed a novel, low-cost and easy-to-implement pretreatment method: "Densifying Lignocellulosic biomass with acidic Chemicals (DLC)" on CS. The DLC-CS owning a uniform shape showed a bulk density 4 times higher compared to loose CS and was highly resistant to microbial contamination, which greatly facilitates biomass handling, transportation and storage. DLC-CS after regular steam autoclave treatment at 121 ○C exhibited high enzymatic digestibility and much higher fermentability compared to traditional dilute acid pretreatment. An ethanol titer as high as 68.1 g/L was achieved without washing or detoxification of the pretreated biomass. The superior performances of DLC for biomass logistics and biomass conversion render it very promising for industrial use. • A novel lignocellulosic biomass pretreatment method was developed. • This method increased biomass bulk density and prevented microbial contamination. • The pretreated biomass showed high digestibility and high fermentability. • A high ethanol titer (68.1 g/L) was achieved without washing or detoxification. • This method facilitates biomass logistics and greatly reduces pretreatment costs. [ABSTRACT FROM AUTHOR]

Abstract Background The growth of the cellulosic ethanol industry is currently impeded by high production costs. One possible solution is to improve the performance of fermentation itself, which has great potential to improve the economics of the entire production process. Here, we demonstrated significantly improved productivity through application of an advanced fermentation approach, named self-cycling fermentation (SCF), for cellulosic ethanol production. Results The flow rate of outlet gas from the fermenter was used as a real-time monitoring parameter to drive the cycling of the ethanol fermentation process. Then, long-term operation of SCF under anaerobic conditions was improved by the addition of ergosterol and fatty acids, which stabilized operation and reduced fermentation time. Finally, an automated SCF system was successfully operated for 21 cycles, with robust behavior and stable ethanol production. SCF maintained similar ethanol titers to batch operation while significantly reducing fermentation and down times. This led to significant improvements in ethanol volumetric productivity (the amount of ethanol produced by a cycle per working volume per cycle time)—ranging from 37.5 to 75.3%, depending on the cycle number, and in annual ethanol productivity (the amount of ethanol that can be produced each year at large scale)—reaching 75.8 ± 2.9%. Improved flocculation, with potential advantages for biomass removal and reduction in downstream costs, was also observed. Conclusion Our successful demonstration of SCF could help reduce production costs for the cellulosic ethanol industry through improved productivity and automated operation.

The article discusses the failure of the United States to meet its goal of producing 16 billion gallons per year of ethanol from cellulosic biomass by 2022. Despite investing $10 billion in public and private funds, less than 10 million gallons of cellulosic ethanol were produced in 2022. The author argues that the failure can be attributed to the lack of five necessary subsystems: large-scale feedstock production, large-scale transport systems, large-scale biorefining systems, large-scale systems to transport biofuels to end markets, and large-scale end markets for the biofuels. The author suggests that the success of renewable natural gas (RNG) produced from waste cellulosic biomass can be attributed to meeting these subsystem requirements, while cellulosic ethanol did not. The article concludes by emphasizing the need to pay attention to these subsystems in order to achieve large-scale cellulosic biofuels production. [Extracted from the article]

Levulinic acid (LA) is one of the most promising biomass-derived platform chemicals owing to its wider convertibility to a large number of commodity chemicals. Numerous LA derivatives have paramount importance in the global economy. In this article, we have comprehensively reviewed various processes that have been developed to produce LA and its derivatives from different sugars and cellulosic feedstocks. These designs are discussed in order to provide comparative information on their chemical mechanisms, process merits, demerits, and scale-up potentials. Monosaccharides such as fructose, glucose, and xylose, and disaccharides such as sucrose are good feedstocks for LA production with Brønsted or Lewis acids as the catalysts in either homogenous or heterogeneous reaction systems. LA yield is in the range of 2–90%, which is greatly dependent on the reaction conditions. Polysaccharides such as cellulose and even lignocellulose are also employed for LA production. Brønsted acids, especially mineral acids, appear to be more efficient than Lewis acids to catalyze the conversion of polysaccharides and lignocellulosic biomass to LA. The important LA derivatives and their preparation reactions such as aminolevulinic acid, diphenolic acid, γ-valerolactone, various alkyl esters, and valerate have also been reviewed. These derivatives have extended utilization in modern industries due to the emergent environmental concerns. Furthermore, the challenges arising during these lab-scale processes are critically unveiled to pave the way for process selection and scale-up study for the production of LA and different derived chemicals. It has been recommended that to improve the production efficiency of LA and its derivatives, efforts should be made to develop robust catalysts and reaction systems in order to improve the reaction selectivity. The mechanisms of LA formation from various feedstocks are also significantly important to guide the intensification of the process. Reactors with potential of industrial application are one of the crucial steps for the scaling up of LA production. [ABSTRACT FROM AUTHOR]

Ethanol to middle distillates (ETMD) is a promising pathway to produce sustainable liquid fuels to decarbonize the hard-to-electrify transportation sectors due to (1) the abundant sugar/starch and lignocellulosic biomass, (2) the existing deployment scale of fuel ethanol production (∼29 billion gallons per year globally), and (3) emerging opportunities in C2+ alcohol synthesis from CO2. Here we report a conceptual market-responsive biorefinery centered around a new ETMD pathway based on one-step ethanol to butene-rich olefins (ETO) over a Cu–Zn–Y/Beta catalyst. Specifically, this ethanol conversion pathway comprises one-step ETO, oligomerization, and hydrotreating. This ETO is distinct from that in the conventional ethanol-to-jet process which is based on two-step ethanol to ethylene and ethylene oligomerization to butenes. Butene-rich olefins can be shifted to butadiene-rich products by simply changing the reaction atmosphere from hydrogen to inert gas over the same ETO catalyst. Leveraging the experimental results, baseline techno-economic analysis (TEA) and sensitivity analysis indicate that the ethanol conversion cost is $0.60 per gallon gasoline equivalent (GGE), with opportunities for further cost reduction via improving the liquid hydrocarbon yield and space velocities, and process optimization on balancing dewatering of ethanol feed prior to the ETO step. The minimum fuel selling price (MFSP) of liquid hydrocarbons derived from corn starch ethanol with butadiene as coproduct is $1.64 per GGE, in the range that is cost competitive with petroleum kerosene-type jet fuel. Projected MFSP for cellulosic ethanol (corn stover) derived hydrocarbons is below $3.00 per GGE and co-production of butadiene further reduces the MFSP to $1.70 per GGE. The Well-to-Wake life-cycle analysis indicates that 85% greenhouse gas emission reduction can be achieved when using corn stover compared to petroleum reference and the associated carbon credits will provide significant economic incentives to favor the cellulosic ethanol-derived hydrocarbon fuels. This study demonstrates a low-cost pathway to middle distillate fuels leveraging existing ethanol infrastructure, where catalysis innovation drives the reduction of process complexity and flexible coproduction of a value-added chemical product. [ABSTRACT FROM AUTHOR]

Kluyveromyces marxianus is an ascomycetous yeast which has shown promising results in cellulosic ethanol and renewable chemicals production. It can survive on a variety of carbon sources under industrially favorable conditions due to its fast growth rate, thermotolerance, and acid tolerance. K. marxianus, is generally regarded as a safe (GRAS) microorganism, is widely recognized as a powerhouse for the production of heterologous proteins and is accepted by the US Food and Drug Administration (USFDA) for its pharmaceutical and food applications. Since lignocellulosic hydrolysates are comprised of diverse monomeric sugars, oligosaccharides and potential metabolism inhibiting compounds, this microorganism can play a pivotal role as it can grow on lignocellulosic hydrolysates coping with vegetal cell wall derived inhibitors. Furthermore, advancements in synthetic biology, for example CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats with Cas9)-mediated genome editing, will enable development of an engineered yeast for the production of biochemicals and biopharmaceuticals having a myriad of industrial applications. Genetic engineering companies such as Cargill, Ginkgo Bioworks, DuPont, Global Yeast, Genomatica, and several others are actively working to develop designer yeasts. Given the important traits and properties of K. marxianus, these companies may find it to be a suitable biocatalyst for renewable chemicals and fuel production on the large scale. This paper reviews the recent progress made with K. marxianus biotechnology for sustainable production of ethanol, and other products utilizing lignocellulosic sugars. [ABSTRACT FROM AUTHOR]

Co-production is a process based on the biorefinery concept that maximizes the benefit of biomass by reusing residue from the production of one product to produce others. In this regard, biogas is one of the most researched second products for the production of ethanol from cellulosic biomass. However, operating this scheme requires additional investment in biogas processing equipment. This review compiles data from research studies on the co-production of bioethanol and biogas from lignocellulosic biomass to determine which is more worthwhile: leaving the residue or investing more to benefit from the second product. According to previous research, ethanol stillage can be converted to biogas via anaerobic digestion, increasing energy output by 2-3 fold. Techno-economic studies demonstrated that the co-production process reduces the minimum ethanol selling price to a level close to the market price of ethanol, implying the possibility of industrializing cellulosic ethanol production through this scheme. [ABSTRACT FROM AUTHOR]

Background: As one of the clean and sustainable energies, lignocellulosic ethanol has achieved much attention around the world. The production of lignocellulosic ethanol does not compete with people for food, while the consumption of ethanol could contribute to the carbon dioxide emission reduction. However, the simultaneous transformation of glucose and xylose to ethanol is one of the key technologies for attaining cost-efficient lignocellulosic ethanol production at an industrial scale. Genetic modification of strains and constructing consortia were two approaches to resolve this issue. Compared with strain improvement, the synergistic interaction of consortia in metabolic pathways should be more useful than using each one separately. Results: In this study, the consortia consisting of suspended Scheffersomyces stipitis CICC1960 and Zymomonas mobilis 8b were cultivated to successfully depress carbon catabolite repression (CCR) in artificially simulated 80G40XRM. With this strategy, a 5.52% more xylose consumption and a 6.52% higher ethanol titer were achieved by the consortium, in which the inoculation ratio between S. stipitis and Z. mobilis was 1:3, compared with the Z. mobilis 8b mono-fermentation. Subsequently, one copy of the xylose metabolic genes was inserted into the Z. mobilis 8b genome to construct Z. mobilis FR2, leading to the xylose final-consumption amount and ethanol titer improvement by 15.36% and 6.81%, respectively. Finally, various corn stover hydrolysates with different sugar concentrations (glucose and xylose 60, 90, 120 g/L), were used to evaluate the fermentation performance of the consortium consisting of S. stipitis CICC1960 and Z. mobilis FR2. Fermentation results showed that a 1.56–4.59% higher ethanol titer was achieved by the consortium compared with the Z. mobilis FR2 mono-fermentation, and a 46.12–102.14% higher ethanol titer was observed in the consortium fermentation when compared with the S. stipitis CICC1960 mono-fermentation. Furthermore, qRT-PCR analysis of xylose/glucose transporter and other genes responsible for CCR explained the reason why the initial ratio inoculation of 1:3 in artificially simulated 80G40XRM had the best fermentation performance in the consortium. Conclusions: The fermentation strategy used in this study, i.e., using a genetically modified consortium, had a superior performance in ethanol production, as compared with the S. stipitis CICC1960 mono-fermentation and the Z. mobilis FR2 mono-fermentation alone. This result showed that this strategy has potential for future lignocellulosic ethanol production. [ABSTRACT FROM AUTHOR]

Background: Hemicellulose acts as one factor contributing to the recalcitrance of lignocellulose that prevents cellulases to degrade the cellulose efficiently even in low quantities. Supplement of hemicellulases can enhance the performance of commercial cellulases in the enzymatic hydrolyses of lignocellulose. Kluyveromyce marxianus is an attractive yeast for cellulosic ethanol fermentation, as well as a promising host for heterologous protein production, since it has remarkable thermotolerance, high growth rate, and broad substrate spectrum etc. In this study, we attempted to coordinately express multiple hemicellulases in K.marxianus through a 2A-mediated ribosome skipping to self-cleave polyproteins, and investigated their capabilities for saccharification and ethanol production from corncobs. Results: Two polycistronic genes IMPX and IMPαX were constructed to test the self-cleavage of P2A sequence from the Foot-and-Mouth Disease virus (FMDV) in K.marxianus. The IMPX gene consisted of a β-mannanase gene M330 (without the stop codon), a P2A sequence and a β-xylanase gene Xyn-CDBFV in turn. In the IMPαX gene, there was an additional α-factor signal sequence in frame with the N-terminus of Xyn-CDBFV. The extracellular β-mannanase activities of the IMPX and IMPαX strains were 21.34 and 15.50 U/mL, respectively, but the extracellular β-xylanase activity of IMPαX strain was much higher than that of the IMPX strain, which was 136.17 and 42.07 U/mL, respectively. Subsequently, two recombinant strains, the IXPαR and IMPαXPαR, were constructed to coordinately and secretorily express two xylantic enzymes, Xyn-CDBFV and β-D-xylosidase RuXyn1, or three hemicellulolytic enzymes including M330, Xyn-CDBFV and RuXyn1. In fed-batch fermentation, extracellular activities of β-xylanase and β-xylosidase in the IXPαR strain were 1664.2 and 0.90 U/mL. Similarly, the IMPαXPαR strain secreted the three enzymes, β-mannanase, β-xylanase, and β-xylosidase, with the activities of 159.8, 2210.5, and 1.25 U/mL, respectively. Hemicellulolases of both strains enhanced the yields of glucose and xylose from diluted acid pretreated (DAP) corncobs when acted synergistically with commercial cellulases. In hybrid saccharification and fermentation (HSF) of DAP corncobs, hemicellulases of the IMPαXPαR strain increased the ethanol yield by 8.7% at 144 h compared with the control. However, both ethanol and xylose yields were increased by 12.7 and 18.2%, respectively, at 120 h in HSF of aqueous ammonia pretreated (AAP) corncobs with this strain. Our results indicated that coordinate expression of hemicellulolytic enzymes in K. marxianus promoted the saccharification and ethanol production from corncobs. Conclusions: The FMDV P2A sequence showed high efficiency in self-cleavage of polyproteins in K. marxianus and could be used for secretory expression of multiple enzymes in the presence of their signal sequences. The IMPαXPαR strain coexpressed three hemicellulolytic enzymes improved the saccharification and ethanol production from corncobs, and could be used as a promising strain for ethanol production from lignocelluloses. [ABSTRACT FROM AUTHOR]

This study investigated cellulosic ethanol production from sugar cane bagasse (SCB) pretreated by triethylbenzyl ammonium chloride/lactic acid (TEBAC/LA) deep eutectic solvent (DES). The results showed that the pretreatment of SCB with TEBAC/LA DES at 120 °C for 4 h with 1:15 (solid to liquid ratio) resulted in the best cellulose digestibility (88.23 ± 1.24%), which was approximately 228% higher than that of untreated SCB. Furthermore, the maximum cellulosic ethanol concentration of 16.84 g/L was achieved using glucose (36.06 g/L) and xylose (7.36 g/L). Moreover, the ethanol productivity and yield were 0.70 g/(L·h) and 0.42 g/g fermentable sugar, respectively. The efficient bioconversion was ascribed to the remarkable delignification (88.72 ± 1.63%), xylan removal (73.93 ± 0.17%), along with optimum cellulose recovery (95.89 ± 1.54%). Importantly, the enzymatic hydrolysis digestibility remained unchanged after 5 times DES recycling process. Overall, it also provided an insight into the efficient SCB biorefinery of DES systems. [Display omitted] [ABSTRACT FROM AUTHOR]

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As recovered from the byproducts stream of a cellulosic ethanol biorefinery, the renewable biopolymer lignin is too impure and polydisperse for many proposed applications. By mixing a hybrid poplar lignin with hot ethanol–water solutions, two liquid phases, one polymer-rich and one solvent-rich, are created. This liquid–liquid equilibrium phenomenon was used to generate solvated (and thus liquefied) lignin fractions of controlled molecular weight for which the impurities analyses for sugars and ash were near or below the limits of detection. Additionally, those carbohydrates and metals impurities end up highly concentrated in a single process stream also having potential value. [ABSTRACT FROM AUTHOR]

Agricultural land and forestland are considered as two largest potential biomass sources for meeting the Renewable Fuel Standard (RFS) mandate for cellulosic biofuels. However, the land use change and greenhouse gas (GHG) savings with both agricultural and forest biomass production are yet to be examined systematically. This paper examines the effects of implementing a 16-billion gallon (60 billion liters) cellulosic biofuel mandate by 2035 on the mix of agricultural and forest biomass, land use change and GHG emissions by using a dynamic partial equilibrium model of the agricultural, forestry and transportation sectors in the US. Our results show that crop residues play a significant role in supplying cellulosic ethanol before 2030, while energy crops are the major feedstocks used for meeting the RFS cellulosic mandate after 2030. Milling and logging residues are economically viable supplements to agricultural biomass for cellulosic ethanol production, though their role in total biomass is small. Across different scenarios of cellulosic ethanol mandate that can be met with either agricultural biomass only or with both agricultural and forest biomass, we find GHG savings from displacing the gasoline range from 0.61 to 0.82 B MgCO2e over the 2015–2035 period. Induced land use change effects associated with expanded feedstock production are modest between and within the agricultural and forestry sectors. We conclude that a mixed feedstock base maximizes the economic and environmental benefits of cellulosic biofuel production. The mitigation potential of cellulosic biofuels is severalfold larger than natural-based solutions such as grassland restoration.

Corncob is a potential feedstock for biorefineries to produce cellulosic ethanol and other chemicals. Densifying lignocellulosic biomass with chemicals followed by autoclave (DLCA) has been confirmed an efficient and economical pretreatment method, and it was applied in the present work for conversion of corncob to bioethanol. The dosage of sulfuric acid, solid loading of biomass, and autoclave time for pretreatment were investigated. Enzymatic hydrolysis at 25–35% solids loadings resulted in 91–97% sugar conversions. Fermentation of the resulted hydrolysates went well with the highest ethanol titer reaching 75.71 g/L at 35% solid loading. Simultaneous saccharification and co-fermentation was applied to further improve xylose consumption at high solids loadings and the ethanol titer was enhanced to 82.0 g/L at 35% solid loading with an ethanol yield of 21.67 kg/100 kg corncob. This study demonstrated DLCA provided a highly digestible and highly fermentable corncob for biorefinery.

Second-generation technology (2G) is currently the industrial route that is being developed with the potential to meet the demand for biofuels and bioproducts. The technology studied in the present work is based on the non-food processing of sugarcane straw, which is unlike the bagasse processing route (a lignocellulosic residue) that is commonly used in boilers for steam and electricity generation. In the last years, cellulosic ethanol plants worldwide include the utilisation of corn stover, sugarcane bagasse, wood waste and plant residues. Therefore, a communication based on industrial aspects using sugarcane straw is important in order to expand the knowledge regarding this promising technology. The main objective of this work included listing the results obtained in a cellulosic-ethanol production industry using sugarcane straw, located in the Northeast region of Brazil and taking into account the efficiency of pre-treatment, enzymatic hydrolysis and fermentation processes and comparing these results with data found in the literature regarding studies with similar efficiency. It was possible to observe that the production of ethanol from sugarcane straw at an industrial scale achieved a productivity of 200 l of ethanol per tonne of biomass, exhibiting 90, 70 and 85% of pre-treatment, hydrolysis and fermentation efficiency, respectively. [ABSTRACT FROM AUTHOR]

One of the most important biofuels is cellulose ethanol which is a popular material for bioethanol production. The present cellulosic ethanol production is through the cellulolytic process and this involves the splitting of complex cellulose into simple sugars through the hydrolysis process of the lignocellulose pretreated with acids and enzymes after which the product is fermented and distilled. There are, however, some challenges due to the enzymatic and acid processes based on the fact that acid hydrolysis has the ability to corrode equipment and cause unwanted waste while the enzymatic hydrolysis process requires a longer time because enzymes are costly and limited. This means there is a need for innovations to minimize the problems associated with these two processes and this led to the application of solid catalysts as the green and effective catalyst to convert cellulose to ethanol. Solid catalysts are resistant to acid and base conditions, have a high surface area, and do not cause corrosion during the conversion of the cellulose due to their neutral pH. This review, therefore, includes the determination of the cellulose potential as feedstock to be used in ethanol production as well as the preparation and application of solid catalyst as the mechanism to convert cellulose into fuel and chemicals. [ABSTRACT FROM AUTHOR]

As a clean energy source, cellulosic ethanol can replace the traditional grain ethanol. If cellulosic ethanol is used widely, it can reduce dependence on fossil fuels. The cost of cellulase is one of the main bottlenecks in the progress of industrialization. How to reduce the cost of enzyme has attracted more attention. Recently, researchers have taken various measures to reduce the cost of cellulase. Enzyme recycling is one of the most efficient ways to reduce cellulase cost of cellulosic ethanol production. The different ways of cellulases recycling were analyzed, on the basis of in-depth analysis of the influence of pretreatment methods on cellulase adsorption, the adsorption mechanism and factors of enzyme and substrate. The cost of enzyme recycling was accounted. Finally, it was found that cellulases recycling had good application potential through cost accounting. [ABSTRACT FROM AUTHOR]

This work presents a study using coconut cultivation wastes as carbon sources to produce lignocellulosic ethanol, such as green coconut shell (GCS), coconut-tree leaflet (CLL) and coconut-tree leaf stalk (CLS). The biomasses were submitted to hydrothermal (HPT) and acidic (APT) pretreatment. Enzymatic hydrolysis was performed using Cellic® Ctec2 supplemented with 10% Cellic® Htec, mainly based on the identification of glucose and xylose. It was found that enzymatic hydrolysis after HPT promoted better results when compared to APT. The glucose concentrations obtained for the biomasses submitted to the HPT were: 31.85 g/L for the GCS, 21.31 g/L for the CLL and 45.39 g/L for the CLS. For xylose, the higher concentration obtained was 7.93 g/L. During the fermentation step applying the yeast Kluyveromyces marxianus, ethanol concentrations (g/L) from HPT and APT hydrolyzed liquors were: 8.83 and 9.71 g/L for GCS, 10.26 and 7.01 g/L for CLL and 12.99 and 7.44 g/L for CLS, respectively. This study identified the energy potential of coconut residues in the production of biofuels, specifically ethanol, from K. marxianus, species which is little explored commercially and able to ferment glucose and xylose. [ABSTRACT FROM AUTHOR]

Technology valuation of cellulosic ethanol production by Kluyveromyces marxianus CCT 7735 based on the concepts of circular economy was investigated. After an acid-base pretreatment followed by saccharification at 50 °C for 72 h, the sweet sorghum hydrolysate was used to produce ethanol at three temperatures: 37, 42, and 45 °C. About 17.83 g/L of cellulosic ethanol was reached at 42 °C in 24 h, with a yield of 2769.8 L/ha sorghum , almost double that obtained for corn straw. This was the best cellulosic ethanol production from sweet sorghum bagasse achieved by K. marxianus strain and, therefore, it was used in the technology valuation. This analysis was carried out through the application of the profit equation, in which the revenue corresponded to ethanol sales in the global market in 2020, and the costs were defined by the activity-based costing method. An integration between 1G and 2G ethanol production, without mixing material streams, was considered to improve the technology valuation analysis. As a result, the equivalent to 17.2% of the profit obtained with the sugarcane juice ethanol was achieved for the sweet sorghum bagasse. Therefore, the technology valuation indicated the potential of sweet sorghum bagasse in the biorefinery ethanol production. [Display omitted] [ABSTRACT FROM AUTHOR]

Cellulosic ethanol is an alternative for increasing the amount of bioethanol production in the world. In Brazil, sugarcane leads the bioethanol production, and to improve its yield, besides bagasse, sugarcane straw is a possible feedstock. However, the process that leads to cell wall disassembly under field conditions is unknown, and understanding how this happens can improve sugarcane biorefinery and soil quality. In the present work, we aimed at studying how sugarcane straw is degraded in the field after 3, 6, 9, and 12 months. Non-structural and structural carbohydrates, lignin content, ash, and cellulose crystallinity were analyzed. The cell wall composition was determined by cell wall fractionation and determination of monosaccharide composition. Non-structural carbohydrates degraded quickly during the first 3 months in the field. Pectins and lignin remained in the plant waste for up to 12 months, while the hemicelluloses and cellulose decreased 7.4 and 12.4%, respectively. Changes in monosaccharide compositions indicated solubilization of arabinoxylan (xylose and arabinose) and β-glucans (β-1,3 1,4 glucan; after 3 months) followed by degradation of cellulose (after 6 months). Despite cellulose reduction, the xylose:glucose ratio increased, suggesting that glucose is consumed faster than xylose. The degradation and solubilization of the cell wall polysaccharides concomitantly increased the level of compounds related to recalcitrance, which led to a reduction in saccharification and an increase in minerals and ash contents. Cellulose crystallinity changed little, with evidence of silica at the latter stages, indicating mineralization of the material. Our data suggest that for better soil mineralization, sugarcane straw must stay in the field for over 1 year. Alternatively, for bioenergy purposes, straw should be used in less than 3 months. [ABSTRACT FROM AUTHOR]

Cellulosic ethanol has attracted much attention as it is of great benefit to social and environmental sustainability. However, the adverse properties of lignocellulosic biomass (i.e. low bulk density, fluffy/difficult to handle, and being easily contaminated by microbes), and issues related to biomass pretreatment (i.e. high energy consumption, difficulty in scale-up and low fermentability of pretreated biomass), largely impede the commercialization of cellulosic ethanol. To address these issues, we invented a novel pretreatment method, Densifying Lignocellulosic biomass with alkaline Chemicals (DLC), and studied DLC on corn stover (CS). DLC-CS has high density and high durability, with contamination prevented and sugar fully preserved, which greatly facilitates biomass logistics. DLC-CS showed high enzymatic digestibility and high fermentability after storage. DLC-CS containing calcium hydroxide yielded 21.4 g of ethanol per 100 g of CS, which was further enhanced to 25.3 g of ethanol per 100 g of CS using a regular steam autoclave. An ethanol titer as high as 70.6 g L−1 was achieved, for the first time, without washing or detoxification of the pretreated biomass. These promising results demonstrated the great potential of DLC pretreatment for lignocellulosic biofuel production. [ABSTRACT FROM AUTHOR]

Cellobiase (E.C. 3.2.1.21) controls the rate limiting step of cellulose hydrolysis in the production of cellulosic ethanol chiefly from agricultural biomass. The effect of thermal stress on secretion of cellobiase by the filamentous fungus Penicillium chrysogenum was studied. Exposure of 40 hours old mycelial culture to 42°C temperature reduced mycelial growth by 10% and resulted in constricted hyphal morphology. The intracellular elucidation of heat stress was also confirmed by upregulation of specific activity of the stress marker protein disulphide isomerase. Concomitantly, heat stress also resulted in increased specific activity of extracellular cellobiase by 50% from 0.5 U/ml to 1 U/ml which was also confirmed through zymography after Native PAGE. The cellobiase retained in intracellular fraction also had higher specific activity indicating that more cellobiase was synthesized under thermal stress. Trehalose is synthesized as a protective osmolyte under physiological stress. However, it is enzymatically degraded as soon as stress subsides. Specific activity of intracellular trehalase slowly increased post stress which indicated gradual dissipation of stress induced trehalose accumulation. The results collectively suggested that the fungus responds to heat stress by resorting to increased production and secretion of cellobiase to mobilize more nutrients from the extracellular environment. The findings provided valuable insights for industrial upregulation of synthesis and secretion of cellobiose. [ABSTRACT FROM AUTHOR]