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The demand for fossil derivate fuels and chemicals has increased, augmenting concerns on climate change, global economic stability, and sustainability on fossil resources. Therefore, the production of fuels and chemicals from alternative and renewable resources has attracted considerable and growing attention. Ethanol is a promising biofuel that can reduce the consumption of gasoline in the transportation sector and related greenhouse gas (GHG) emissions. Lignocellulosic biomass is a promising feedstock to produce bioethanol (cellulosic ethanol) because of its abundance and low cost. Since the conversion of lignocellulose to ethanol is complex and expensive, the cellulosic ethanol price cannot compete with those of the fossil derivate fuels. A promising strategy to lower the production cost of cellulosic ethanol is developing a biorefinery which produces ethanol and other high-value chemicals from lignocellulose. The selection of such chemicals is difficult because there are hundreds of products that can be produced from lignocellulose. Multiple reviews and reports have described a small group of lignocellulose derivate compounds that have the potential to be commercialized. Some of these products are in the bench scale and require extensive research and time before they can be industrially produced. This review examines chemicals and materials with a Technology Readiness Level (TRL) of at least 8, which have reached a commercial scale and could be shortly or immediately integrated into a cellulosic ethanol process.

Lignocellulosic biomass (LCB) is globally available and sustainable feedstock containing sugar-rich platform that can be converted to biofuels and specialty products through appropriate processing. This review focuses on the efforts required for the development of sustainable and economically viable lignocellulosic biorefinery to produce carbon neutral biofuels along with the specialty chemicals. Sustainable biomass processing is a global challenge that requires the fulfillment of fundamental demands concerning economic efficiency, environmental compatibility, and social responsibility. The key technical challenges in continuous biomass supply and the biological routes for its saccharification with high yields of sugar sources have not been addressed in research programs dealing with biomass processing. Though many R&D endeavors have directed towards biomass valorization over several decades, the integrated production of biofuels and chemicals still needs optimization from both technical and economical perspectives. None of the current pretreatment methods has advantages over others since their outcomes depend on the type of feedstock, downstream process configuration, and many other factors. Consolidated bio-processing (CBP) involves the use of single or consortium of microbes to deconstruct biomass without pretreatment. The use of new genetic engineering tools for natively cellulolytic microbes would make the CBP process low cost and ecologically friendly. Issues arising with chemical characteristics and rigidity of the biomass structure can be a setback for its viability for biofuel conversion. Integration of functional genomics and system biology with synthetic biology and metabolic engineering undoubtedly led to generation of efficient microbial systems, albeit with limited commercial potential. These efficient microbial systems with new metabolic routes can be exploited for production of commodity chemicals from all the three components of biomass. This paper provides an overview of the challenges that are faced by the processes converting LCB to commodity chemicals with special reference to biofuels.

Great effort has been devoted to engineering Saccharomyces cerevisiae with pentose metabolism through the oxido-reductase pathway for cellulosic ethanol production, but intrinsic cofactor imbalance is observed, which substantially compromises ethanol yield. Zymomonas mobilis not only can be engineered for pentose metabolism through the isomerase pathway without cofactor imbalance but also metabolizes sugar through the Entner–Doudoroff pathway with less ATP and biomass produced for more sugar to be used for ethanol production. Moreover, the availabilities of genome sequence information for multiple Z. mobilis strains and advanced genetics tools have laid a solid foundation for engineering this species, and the self-flocculation of the bacterial cells also presents significant advantages for bioprocess engineering. Here, we highlight some of recent advances in these aspects. Zymomonas mobilis metabolizes glucose through the Entner–Doudoroff (ED) pathway, with less ATP generated and biomass accumulated for more ethanol production. The large specific cell surface of Z. mobilis together with the ED pathway facilitates glucose uptake and ethanol fermentation. Its metabolic characteristics and narrow substrate spectrum make Z. mobilis unsuitable for fuel ethanol production from sugar- and starch-based feedstocks, but it would be a good host to be engineered for cellulosic ethanol production. When self-flocculated, Z. mobilis can be immobilized within fermenters for high cell density to improve ethanol productivity. Meanwhile, its tolerance to environmental stresses may be enhanced by this morphological change. Both Z. mobilis ZM4 and its self-flocculating mutant ZM401 can tolerate more than 100g/L ethanol, which is sufficient for cellulosic ethanol production.

The industrial production of sugar syrups from lignocellulosic materials requires the conduction of the enzymatic hydrolysis step at high-solids loadings (i.e., with over 15% solids [w/w] in the reaction mixture). Such conditions result in sugar syrups with increased concentrations and in improvements in both capital and operational costs, making the process more economically feasible. However, this approach still poses several technical hindrances that impact the process efficiency, known as the “high-solids effect” (i.e., the decrease in glucan conversion yields as solids load increases). The purpose of this review was to present the findings on the main limitations and advances in high-solids enzymatic hydrolysis in an updated and comprehensive manner. The causes for the rheological limitations at the onset of the high-solids operation as well as those influencing the “high-solids effect” will be discussed. The subject of water constraint, which results in a highly viscous system and impairs mixing, and by extension, mass and heat transfer, will be analyzed under the perspective of the limitations imposed to the action of the cellulolytic enzymes. The “high-solids effect” will be further discussed vis-à-vis enzymes end-product inhibition and the inhibitory effect of compounds formed during the biomass pretreatment as well as the enzymes’ unproductive adsorption to lignin. This review also presents the scientific and technological advances being introduced to lessen high-solids hydrolysis hindrances, such as the development of more efficient enzyme formulations, biomass and enzyme feeding strategies, reactor and impeller designs as well as process strategies to alleviate the end-product inhibition. We surveyed the academic literature in the form of scientific papers as well as patents to showcase the efforts on technological development and industrial implementation of the use of lignocellulosic materials as renewable feedstocks. Using a critical approach, we expect that this review will aid in the identification of areas with higher demand for scientific and technological efforts.

Yeast to directly convert cellulose and, especially, the microcrystalline cellulose into bioethanol, was engineered through display of minicellulosomes on the cell surface of Saccharomyces cerevisiae . The construction and cell surface attachment of cellulosomes were accomplished with two individual miniscaffoldins to increase the display level. All of the cellulases including a celCCA (endoglucanase), a celCCE (cellobiohydrolase), and a Ccel_2454 (β-glucosidase) were cloned from Clostridium cellulolyticum , ensuring the thermal compatibility between cellulose hydrolysis and yeast fermentation. Cellulases and one of miniscaffoldins were secreted by α-factor; thus, the assembly and attachment to anchoring miniscaffoldin were accomplished extracellularly. Immunofluorescence microscopy, flow cytometric analysis (FACS), and cellulosic ethanol fermentation confirmed the successful display of such complex on the yeast surface. Enzyme–enzyme synergy, enzyme-proximity synergy, and cellulose–enzyme–cell synergy were analyzed, and the length of anchoring miniscaffoldin was optimized. The engineered S. cerevisiae was applied in fermentation of carboxymethyl cellulose (CMC), phosphoric acid-swollen cellulose (PASC), or Avicel. It showed a significant hydrolytic activity toward microcrystalline cellulose, with an ethanol titer of 1,412 mg/L. This indicates that simultaneous saccharification and fermentation of crystalline cellulose to ethanol can be accomplished by the yeast, engineered with minicellulosome.

Switchgrass is a leading dedicated bioenergy feedstock in the United States because it is a native, high-yielding, perennial prairie grass with a broad cultivation range and low agronomic input requirements. Biomass conversion research has developed processes for production of ethanol and other biofuels, but they remain costly primarily because of the intrinsic recalcitrance of biomass. We show here that genetic modification of switchgrass can produce phenotypically normal plants that have reduced thermal-chemical ([less-than or equal to]180 °C), enzymatic, and microbial recalcitrance. Down-regulation of the switchgrass caffeic acid O-methyltransferase gene decreases lignin content modestly, reduces the syringyl:guaiacyl lignin monomer ratio, improves forage quality, and, most importantly, increases the ethanol yield by up to 38% using conventional biomass fermentation processes. The down-regulated lines require less severe pretreatment and 300-400% lower cellulase dosages for equivalent product yields using simultaneous saccharification and fermentation with yeast. Furthermore, fermentation of diluted acid-pretreated transgenic switchgrass using Clostridium thermocellum with no added enzymes showed better product yields than obtained with unmodified switchgrass. Therefore, this apparent reduction in the recalcitrance of transgenic switchgrass has the potential to lower processing costs for biomass fermentation-derived fuels and chemicals significantly. Alternatively, such modified transgenic switchgrass lines should yield significantly more fermentation chemicals per hectare under identical process conditions.

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.

Background During the past few years, the first industrial-scale cellulosic ethanol plants have been inaugurated. Although the performance of the commercial cellulase enzymes used in this process has greatly improved over the past decade, cellulases still represent a very significant operational cost. Depending on the region, transport of cellulases from a central production facility to a biorefinery may significantly add to enzyme cost. The aim of the present study was to develop a simple, cost-efficient cellulase production process that could be employed locally at a Brazilian sugarcane biorefinery. Results Our work focused on two main topics: growth medium formulation and strain improvement. We evaluated several Brazilian low-cost industrial residues for their potential in cellulase production. Among the solid residues evaluated, soybean hulls were found to display clearly the most desirable characteristics. We engineered a Trichoderma reesei strain to secrete cellulase in the presence of repressing sugars, enabling the use of sugarcane molasses as an additional carbon source. In addition, we added a heterologous β-glucosidase to improve the performance of the produced enzymes in hydrolysis. Finally, the addition of an invertase gene from Aspegillus niger into our strain allowed it to consume sucrose from sugarcane molasses directly. Preliminary cost analysis showed that the overall process can provide for very low-cost enzyme with good hydrolysis performance on industrially pre-treated sugarcane straw. Conclusions In this study, we showed that with relatively few genetic modifications and the right growth medium it is possible to produce considerable amounts of well-performing cellulase at very low cost in Brazil using T. reesei. With further enhancements and optimization, such a system could provide a viable alternative to delivered commercial cellulases.

Globally, bioethanol is the largest volume biofuel used in the transportation sector, with corn-based ethanol production occurring mostly in the US and sugarcane-based ethanol production occurring mostly in Brazil. Advances in technology and the resulting improved productivity in corn and sugarcane farming and ethanol conversion, together with biofuel policies, have contributed to the significant expansion of ethanol production in the past 20 years. These improvements have increased the energy and greenhouse gas (GHG) benefits of using bioethanol as opposed to using petroleum gasoline. This article presents results from our most recently updated simulations of energy use and GHG emissions that result from using bioethanol made from several feedstocks. The results were generated with the GREET (Greenhouse gases, Regulated Emissions, and Energy use in Transportation) model. In particular, based on a consistent and systematic model platform, we estimate life-cycle energy consumption and GHG emissions from using ethanol produced from five feedstocks: corn, sugarcane, corn stover, switchgrass and miscanthus. We quantitatively address the impacts of a few critical factors that affect life-cycle GHG emissions from bioethanol. Even when the highly debated land use change GHG emissions are included, changing from corn to sugarcane and then to cellulosic biomass helps to significantly increase the reductions in energy use and GHG emissions from using bioethanol. Relative to petroleum gasoline, ethanol from corn, sugarcane, corn stover, switchgrass and miscanthus can reduce life-cycle GHG emissions by 19-48%, 40-62%, 90-103%, 77-97% and 101-115%, respectively. Similar trends have been found with regard to fossil energy benefits for the five bioethanol pathways.

Ethanol production from cellulosic material is considered one of the most promising options for future biofuel production contributing to both the energy diversification and decarbonization of the transport sector, especially where electricity is not a viable option (e.g., aviation). Compared to conventional (or first generation) ethanol production from food and feed crops (mainly sugar and starch based crops), cellulosic (or second generation) ethanol provides better performance in terms of greenhouse gas (GHG) emissions savings and low risk of direct and indirect land-use change. However, despite the policy support (in terms of targets) and significant R&D funding in the last decade (both in EU and outside the EU), cellulosic ethanol production appears to be still limited. The paper provides a comprehensive overview of the status of cellulosic ethanol production in EU and outside EU, reviewing available literature and highlighting technical and non-technical barriers that still limit its production at commercial scale. The review shows that the cellulosic ethanol sector appears to be still stagnating, characterized by technical difficulties as well as high production costs. Competitiveness issues, against standard starch based ethanol, are evident considering many commercial scale cellulosic ethanol plants appear to be currently in idle or on-hold states.