Butanol Production Research Articles

Capability of Immobilized Clostridium beijerinckii TISTR 1461 on Lotus Stalk Pieces to Produce Butanol from Sugarcane Molasses

Immobilized Clostridium beijerinckii TISTR 1461 was used to enhance the butanol production efficiency from sugarcane molasses. Lotus stalk (LS) pieces were used as carriers for cell immobilization. Sugarcane molasses containing 50 g/L of sugar supplemented with 1 g/L of yeast extract was found to be an appropriate medium for bacterial cell immobilization on the LS pieces. Carrier size (4, 12 and 20 mm in length) and carrier loading (1:15, 1:30 and 1:45, w/v) were optimized for high levels of butanol production using response surface methodology (RSM). The batch fermentation was carried out under anaerobic conditions in 1 L screw-capped bottles at 37 °C and an agitation rate of 150 rpm. It was found that the optimum conditions for the butanol production were the carrier size of 4 mm and carrier loading of 1:31 (w/v). Under these conditions, the butanol concentration (PB) was 12.89 g/L, corresponding to the butanol productivity (QB) of 0.36 g/L∙h and butanol yield (YB/S) of 0.36 g/g. These values were higher than those using free cells (PB, 10.20 g/L, QB, 0.28 g/L∙h and YB/S, 0.32 g/g). In addition, it was found that a 24 h incubation time for cell immobilization was appropriate for the immobilization process, which was confirmed by the results of the scanning electron microscope (SEM) and atomic force microscopy (AFM) images and specific surface area measurement. When the fermentation using the immobilized cells was carried out in a stirred-tank reactor (STR), column reactor (CR) and CR coupled with STR, the results showed that all reactors could be used to produce butanol production from the immobilized cells on LS pieces. However, the PB using CR and CR coupled with STR were only 75% and 45% of those using the screw-capped bottle and STR.

Sustainable Purification of Butanol from a Class of a Mixture Produced by Reduction of Volatile Fatty Acids

There has been an increasing interest in the conversion of biomass to biofuels, energy, and chemicals due to an increase in meeting environmental demands and price and decrease in the potential availability of crude oil. Among the biofuels postulated as viable alternatives due to their physicochemical characteristics is butanol. Given its high energy content, it is projected as a potential substitute for ordinary gasoline. However, butanol production process through fermentation of lignocellulosic material has shown some disadvantages. Another way of producing butanol is by reduction of volatile fatty acids (from waste streams of organic matters) with hydrogen. An effluent with a high content of water and butanol is obtained. In that sense, thermodynamic interactions make the separation process challenging. On the other hand, current policies and needs have guided the proposals for chemical processes to meet various sustainability metrics, for example, high profit margins and low environmental impact, with inherent safety and robust operation in the presence of disturbances. With this in mind, this work proposes purification schemes to obtain butanol of high purity, from a butanol–water mixture, in the compositions generated by reduction of volatile fatty acids, using pervaporation, pressure swing distillation, and azeotropic distillation. Comparing the results obtained, the pervaporation scheme turned out to be the most promising alternative as it presents reductions in all the “green” indicators (compared to the other purification alternatives) in percentages between 27 and 52%. The general indices for such alternative were 0.0392 ($/kgbutanol), 0.0066 (ecopoints/kgbutanol), 8274, 2.772 × 10–04 (probability/year), and 0.4281 $/kgbutanol regarding the total annual cost, ecological indicator 99, condition number, individual risk, and minimum selling price, respectively.

Orange peel extract enhanced sugar recovery and butanol production from potato peel by Clostridium acetobutylicum

ABSTRACT The present study is focused on biobutanol production from potato peel waste in dilute orange peel extract by Clostridium acetobutylicum MTCC 11274 at three concentrations of 20, 40, and 60 g/L as sole nutrient sources. The biomass and solvent production from potato peel were enhanced in orange peel extract with the concentration and improved the sugar recovery. The effect on solvent production from 60 g/L potato peel in dilute orange peel extract was relatively higher than 20 and 40 g/L. The butanol yield was improved from 0.04 to 0.35 g/g from 60 g/L potato peel waste in orange peel extract. The butyric acid concentration was highly correlated with butanol production. The Clostridium acetobutylicum MTCC 11274 showed no inhibitory effect from D-limonene in orange peel extract. The potato peel waste hydrolyzate prepared in orange peel extract enhanced the growth of Clostridium acetobutylicum MTCC 11274 and fermentation of acetone, butanol, and ethanol.

Identification and Investigation of Autolysin Genes in Clostridium saccharoperbutylacetonicum Strain N1-4 for Enhanced Biobutanol Production.

Biobutanol is a valuable biochemical and one of the most promising biofuels. Clostridium saccharoperbutylacetonicum N1-4 is a hyperbutanol-producing strain. However, its strong autolytic behavior leads to poor cell stability, especially during continuous fermentation, thus limiting the applicability of the strain for long-term and industrial-scale processes. In this study, we aimed to evaluate the role of autolysin genes within the C. saccharoperbutylacetonicum genome related to cell autolysis and further develop more stable strains for enhanced butanol production. First, putative autolysin-encoding genes were identified in the strain based on comparison of amino acid sequence with homologous genes in other strains. Then, by overexpressing all these putative autolysin genes individually and characterizing the corresponding recombinant strains, four key genes were pinpointed to be responsible for significant cell autolysis activities. Further, these key genes were deleted using CRISPR-Cas9. Fermentation characterization demonstrated enhanced performance of the resultant mutants. Results from this study reveal valuable insights concerning the role of autolysins for cell stability and solvent production, and they provide an essential reference for developing robust strains for enhanced biofuel and biochemical production.IMPORTANCE Severe autolytic behavior is a common issue in Clostridium and many other microorganisms. This study revealed the key genes responsible for the cell autolysis within Clostridium saccharoperbutylacetonicum, a prominent platform for biosolvent production from lignocellulosic materials. The knowledge generated in this study provides insights concerning cell autolysis in relevant microbial systems and gives essential references for enhancing strain stability through rational genome engineering.

Aqueous pretreatment of triticale straw for integrated production of hemicellulosic methane and cellulosic butanol

Despite its advantages, lignocellulosic butanol cannot become a real alternative without processes for obtaining credits from non-cellulosic fraction of lignocellulose. In this study, the process of “cellulosic butanol” was integrated with “hemicellulosic methane” process, i.e., anaerobic digestion (AD), for higher energy return on investment (EROI). Aqueous pretreatment was evaluated as a connecting chain between the processes. The alkaline (1% NaOH), acidic (1% H2SO4), and neutral (autohydrolysis) pretreatments were utilized at 140, 160, and 180 °C for bioenergy recovery from triticale straw. Enzymatic hydrolysis of the straw pretreated by alkaline (180 °C), dilute acid (140 °C), and autohydrolysis (180 °C) and fermentation of the hydrolysates by Clostridium acetobutylicum led to 13.6, 8.5, and 10.2 g/L acetone-butanol-ethanol (ABE), respectively. AD of the liquors remained after pretreatment by alkaline, dilute acid, and autohydrolysis led to 83–121, 240–247, and 380–430 mL/g VSS methane leading to more than 60% increase in EROI.

Butyrate production in the acetogen Eubacterium limosum is dependent on the carbon and energy source.

Summary Eubacterium limosum KIST612 is one of the few acetogenic bacteria that has the genes encoding for butyrate synthesis from acetyl‐CoA, and indeed, E. limosum KIST612 is known to produce butyrate from CO but not from H2 + CO2. Butyrate production from CO was only seen in bioreactors with cell recycling or in batch cultures with addition of acetate. Here, we present detailed study on growth of E. limosum KIST612 on different carbon and energy sources with the goal, to find other substrates that lead to butyrate formation. Batch fermentations in serum bottles revealed that acetate was the major product under all conditions investigated. Butyrate formation from the C1 compounds carbon dioxide and hydrogen, carbon monoxide or formate was not observed. However, growth on glucose led to butyrate formation, but only in the stationary growth phase. A maximum of 4.3 mM butyrate was observed, corresponding to a butyrate:glucose ratio of 0.21:1 and a butyrate:acetate ratio of 0.14:1. Interestingly, growth on the C1 substrate methanol also led to butyrate formation in the stationary growth phase with a butyrate:methanol ratio of 0.17:1 and a butyrate:acetate ratio of 0.33:1. Since methanol can be produced chemically from carbon dioxide, this offers the possibility for a combined chemical‐biochemical production of butyrate from H2 + CO2 using this acetogenic biocatalyst. With the advent of genetic methods in acetogens, butanol production from methanol maybe possible as well.

Acetone, Butanol, Ethanol and, Xylitol Production Through a Biorefinery Platform: An Experimental & Simulation Approach

Butanol is an interesting biofuel and a product precursor, that could be obtained with acetone and ethanol via fermentation. The biofuels production has been identified as not economically competitive, thus, the parallel production of high value-added products, such as xylitol, could be an alternative to improve the profit. Xylitol can be produced from xylose, which might be considered as a coproduct in a second generation biorefinery. This study presents a systematic biorefinery process design for the simultaneous acetone, butanol, ethanol (ABE) and xylitol production, based on experimental and simulation approaches. Experiments were performed for the pretreatment of sugarcane bagasse and ABE fermentation. The simulation part used the experimental results and experimental data from literature, to perform rigorous calculations of the ABE and xylitol production process. The economic analysis (EA) was performed relying on some indicators such as, the net present value (NPV) and payback period (PBP); EA includes several scenarios for producing only ABE and some scenarios for simultaneous ABE and xylitol production. The results showed that the combined butanol and xylitol production could reduce by 17% the selling price of butanol, compared with only producing butanol. The study also included the combustion of residual solids and carbon dioxide depletion analyses. This approach illustrates the opportunity to perform a rigorous techno-economic analysis, to identify the feasibility of the process at industrial scale, based on realistic data. This approach was implemented for ABE and xylitol production, but it can be used to any other bioproduct.

A comparative phenotypic and genomic analysis of Clostridium beijerinckii mutant with enhanced solvent production

The acetone-butanol-ethanol (ABE) fermentation by solventogenic clostridia has a long history of industrial butanol production. The Clostridium beijerinckii mutant BA101 has been widely studied for ABE fermentation owing to its enhanced butanol production capacity. Here, we characterized the BA101 mutant under controlled environmental conditions in parallel with the parental strain C. beijerinckii NCIMB 8052. To investigate the correlation between phenotype and genotype, we carried out the genome sequencing of BA101. Through comparative genomic analysis, several mutations in the genes encoding transcriptional regulator, sensor kinase, and phosphatase were identified in the BA101 genome as well as other sibling mutants. Among them, the SNP in the Cbei_3078 gene encoding PAS/PAC sensor hybrid histidine kinase was unique to the BA101 strain. The identified mutations relevant to the observed physiological behaviors of BA101 could be potential genetic targets for rational engineering of solventogenic clostridia toward desired phenotypes.

Novel Methods Using an Arthrobacter sp. to Create Anaerobic Conditions for Biobutanol Production from Sweet Sorghum Juice by Clostridium beijerinckii

Biobutanol can be produced by Clostridia via an acetone–butanol–ethanol (ABE) fermentation under strictly anaerobic conditions. Oxygen-free nitrogen (OFN) gas is typically used to create anaerobic conditions for ABE fermentations. However, this method is not appropriate for large-scale fermentations as it is quite costly. The aim of this work was to study the feasibility of butanol production from sweet sorghum juice (SSJ) by Clostridium beijerinckii TISTR 1461 using various methods to create anaerobic conditions, i.e., growth of a strictly aerobic bacterium, an Arthrobacter sp., under different conditions and a chemical method using sodium dithionite (SDTN) to consume residual oxygen. SSJ containing 60 g/L of total sugar supplemented with 1.27 g/L of (NH4)2SO4 was used as a substrate for butanol production. The results showed that 0.25 mM SDTN could create anaerobic conditions, but in this case, C.beijerinckii TISTR 1461 could produce butanol at a concentration (PB) of only 8.51 g/L with a butanol productivity (QB) of 0.10 g/L·h. Arthrobacter sp. BCC 72131 could also be used to create anaerobic conditions. Mixed cultures of C.beijerinckii TISTR 1461 and Arthrobacter sp. BCC 72131 created anaerobic conditions by inoculating the C.beijerinckii 4 h after Arthrobacter. This gave a PB of 10.39 g/L with a QB of 0.20 g/L·h. Comparing butanol production with the control treatment (using OFN gas to create anaerobic conditions, yielding a PB of 9.88 g/L and QB of 0.21 g/L·h) indicated that using Arthrobacter sp. BCC 72131 was an appropriate procedure for creating anaerobic conditions for high levels of butanol production by C. beijerinckii TISTR 1461 from a SSJ medium.

Quantitative proteomic analysis to reveal expression differences for butanol production from glycerol and glucose by Clostridium sp. strain CT7

Clostridium sp. strain CT7 is a new emerging microbial cell factory with high butanol production ratio owing to its non-traditional butanol fermentation mode with uncoupled acetone and 1,3-propanediol formation. Significant changes of metabolic products profile were shown in glycerol- and glucose-fed strain CT7, especially higher butanol and lower volatile fatty acids (VFAs) production occurred from glycerol-fed one. However, the mechanism of this interesting phenomenon was still unclear. To better elaborate the bacterial response towards glycerol and glucose, the quantitative proteomic analysis through iTRAQ strategy was performed to reveal the regulated proteomic expression levels under different substrates. Proteomics data showed that proteomic expression levels related with carbon metabolism and solvent generation under glycerol media were highly increased. In addition, the up-regulation of hydrogenases, ferredoxins and electron-transferring proteins may attribute to the internal redox balance, while the earlier triggered sporulation response in glycerol-fed media may be associated with the higher butanol production. This study will pave the way for metabolic engineering of other industrial microorganisms to obtain efficient butanol production from glycerol.

Energy-efficient butanol production by Clostridium acetobutylicum with histidine kinase knockouts to improve strain tolerance and process robustness

Engineering histidine kinases in C. acetobutylicum enhanced cell viability and solventogenesis in ABE fermentation and enabled robust and energy-efficient butanol production.

Efficient Strategy to Alleviate the Inhibitory Effect of Lignin-Derived Compounds for Enhanced Butanol Production

In the present study, the effect of one of the most important lignin-derived inhibitors (lignosulfonate) was assessed. A technique to overcome the lignosulfonate inhibitory action in the acetone–bu...

The combined effect on initial glucose concentration and pH control strategies for acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum DSM 792

The use and depletion of fossil fuels raised the interest in biofuels like biobutanol. Clostridium acetobutylicum DSM 792 is capable of producing biobutanol through ABE fermentation. Butanol production can be influenced by low sugar concentrations, like those obtained after hydrolysis of pre-treated lignocellulosic biomass. This study aimed to evaluate the influence of the initial glucose concentrations (33, 66 and 100 g L−1) and pH control strategies on biobutanol production and glucose consumption. Uncontrolled pH fermentation exhibited low butanol production due to either glucose exhaustion (33 g L−1) or the phenomenon of acid crash (66 and 100 g L−1), which was alleviated by the use of any of the minimum pH set-points (4.8; 5.0; 5.1 and 5.5). Fermentation at a pHmin of 5.1 gave the best performance in butanol production and glucose consumption rate. Fermenting with a pHmin of 5.1 with 33 g L−1 of initial glucose caused acidogenic fermentation, while the use of 66 and 100 g L−1 of glucose led to a ∼1.5 and ∼1.7-fold increase in butanol concentration over their counterparts without pH control respectively. By controlling the pH after the acidogenic phase 15.8 g L−1 of butanol was obtained with 100 g L−1 of glucose, which was ∼2-fold higher than without pH control. Different initial glucose concentrations therefore require different pH strategies to optimize butanol production.

Effects of Carbon Ion Beam Irradiation on Butanol Tolerance and Production of Clostridium acetobutylicum.

Clostridium acetobutylicum (C. acetobutylicum) has considerable potential for use in bioenergy development. Owing to the repeated use of traditional mutagenesis methods, the strains have developed a certain tolerance. The rheology of the bioprocess and the downstream processing of the product heavily depend on the ability of C. acetobutylicum mutants to produce butanol. Carbon ion beam irradiation has advantages over traditional mutation methods for fermentative production because of its dose conformity and superb biological effectiveness. However, its effects on the specific productivity of the strains have not been clearly understood. In this study, we screened five mutants through carbon ion beam irradiation; mutant Y217 achieved a butanol-production level of 13.67 g/L, exceeding that of wild-type strain ATCC 824 (i.e., 9.77 g/L). In addition, we found that the mutant maintained normal cell membrane integrity under the stimulation of 15 g/L butanol, whereas the intracellular macromolecules of wild-type strain ATCC 824 leaked significantly. Subsequently, we used the response surface methodology (RSM) to determine if the mutant cell membrane integrity improved the butanol tolerance. We verified that with the addition of butanol, the mutant could be fermented to produce 8.35 g/L butanol, and the final butanol concentration in the fermentation broth could reach 16.15 g/L. In this study, we proved that under butanol stress, mutant Y217 features excellent butanol production and tolerance and cell membrane integrity and permeability; no prior studies have attempted to do so. This will serve as an interesting and important illustration of the complexity of genetic control of the irradiation mutation of C. acetobutylicum strains. It may also prove to be useful in the bioengineering of strains of the mutant for use in the predevelopment stage.

Inhibitors tolerance analysis of Clostridium sp. strain LJ4 and its application for butanol production from corncob hydrolysate through electrochemical detoxification

Lignocellulose is a promising feedstock for butanol production owing to its abundance and renewability, however, most solventogenic clostridia could not directly convert these low-cost resources. Fermentable sugars could only be produced through pretreatment and hydrolysis processes, but furan and phenolic derivatives occurring in the lignocellulosic hydrolysate would inhibit cells’ growth and final butanol production capability. Compared to reported solventogenic Clostridia, the newly isolated Clostridium sp. strain LJ4 possessed high tolerance towards furan derivatives. Genes of sdr4, sdr1, akr, fmn and ad2 in strain LJ4 may play key roles in the bioconversion of furan derivatives based on the transcriptional levels’ analysis. As the phenolic derivatives containing in the undetoxified corncob hydrolysate were higher than the limited tolerance of strain LJ4, an electrochemical detoxification method was accordingly adopted to reduce the toxicity of phenolic derivatives without the loss of reducing sugars and generation of by-products. After 24 h detoxification of acid treated corncob hydrolysate, the phenolic compounds were decreased by nearly 40%, and 4.7 g/L of butanol was finally produced by strain LJ4 with 1.6-fold improvement than that from the un-detoxified corncob hydrolysate. Electrochemical technique could become a promising approach for biofuels or biochemicals production from undetoxified lignocellulosic hydrolysate owing to its high removal efficiency of phenolic derivatives without sugar loss.

Enhanced coproduction and trade-off of the hydrogen and butanol in the coupled system of pervaporation and repeated-cycle fixed-bed fermentation

In the study, a coupled system of pervaporation and repeated-cycle fixed bed fermentation with the bagasse as the carrier material was developed. Three repeated cycles of fermentation lasting for 540 h was efficiently carried out to coproduce hydrogen and butanol with the strain Clostridium acetobutylicum. From 1st cycle to 3rd cycle, the average hydrogen productivity and butanol productivity were around 130 mLL−1 h−1 and 0.15 gL−1 h−1, respectively. Average hydrogen yield and butanol yield were around 0.17 Lg−1 and 0.19 gg−1. The hydrogen and butanol production per unit cell weight were 27 Lg−1 and 31 gg−1, which were more than two times of those in a conventional repeated-cycle fermentation. The trade-off and competition between the hydrogen and butanol production was studied and analyzed. Compared with the pervaporation-free fermentation, the hydrogen yield was decreased by 14 % and butanol yield was increased by 27 % in the coupled system. Instantaneous solvent removal from the broth by pervaporation changed probably the metabolic pathway to more butanol production over hydrogen. From 1st cycle to 3rd cycle, the respiration heat generated during each fermentation cycle could cover 45 %, 24 % and 37 % of the total energy requirement of the pervaporation achieving the best use of the energy.

Deeper below the surface-transcriptional changes in selected genes of Clostridium beijerinckii in response to butanol shock.

The main bottleneck in the return of industrial butanol production from renewable feedstock through acetone–butanol–ethanol (ABE) fermentation by clostridia, such as Clostridium beijerinckii, is the low final butanol concentration. The problem is caused by the high toxicity of butanol to the production cells, and therefore, understanding the mechanisms by which clostridia react to butanol shock is of key importance. Detailed analyses of transcriptome data that were obtained after butanol shock and their comparison with data from standard ABE fermentation have resulted in new findings, while confirmed expected population responses. Although butanol shock resulted in upregulation of heat shock protein genes, their regulation is different than was assumed based on standard ABE fermentation transcriptome data. While glucose uptake, glycolysis, and acidogenesis genes were downregulated after butanol shock, solventogenesis genes were upregulated. Cyclopropanation of fatty acids and formation of plasmalogens seem to be significant processes involved in cell membrane stabilization in the presence of butanol. Surprisingly, one of the three identified Agr quorum‐sensing system genes was upregulated. Upregulation of several putative butanol efflux pumps was described after butanol addition and a large putative polyketide gene cluster was found, the transcription of which seemed to depend on the concentration of butanol.

Three-stage repeated-batch immobilized cell fermentation to produce butanol from non-detoxified sugarcane bagasse hemicellulose hydrolysates

To enable the production of butanol with undiluted, non-detoxified sugarcane bagasse hemicellulose hydrolysates, this study developed a three-staged repeated-batch immobilized cell fermentation in which the efficiency of a 3D-printed nylon carrier to passively immobilize Clostridium saccharoperbutylacetonicum DSM 14923 was compared with sugarcane bagasse. The first stage consisted of sugarcane molasses fermentation, and in the second stage, non-detoxified sugarcane bagasse hemicellulose hydrolysates (SBHH) was pulse-fed to sugarcane molasses fermentation. In the next four batches, immobilized cells were fed with undiluted SBHH supplemented with molasses, and SBHH-derived xylose accounted for approximately 50% of the sugars. Bagasse was a superior carrier, and the average xylose utilization (33%) was significantly higher than the treatment with the 3D-printed carrier (16%). Notably, bagasse allowed for 43% of the butanol to be SBHH-derived. Overall, cell immobilization on lignocellulosic materials can be an efficient strategy to produce butanol from repeated-batch fermentation of non-detoxified hemicellulose hydrolysates.

Metabolomic and kinetic investigations on the electricity-aided production of butanol by Clostridium pasteurianum strains.

In this contribution, we studied the effect of electro‐fermentation on the butanol production of Clostridium pasteurianum strains by a targeted metabolomics approach. Two strains were examined: an electrocompetent wild type strain (R525) and a mutant strain (dhaB mutant) lacking formation of 1,3‐propanediol (PDO). The dhaB‐negative strain was able to grow on glycerol without formation of PDO, but displayed a high initial intracellular NADH/NAD ratio which was lowered subsequently by upregulation of the butanol production pathway. Both strains showed a 3–5 fold increase of the intracellular NADH/NAD ratio when exposed to cathodic current in a bioelectrochemical system (BES). This drove an activation of the butanol pathway and resulted in a higher molar butanol to PDO ratio for the R525 strain. Nonetheless, macroscopic electron balances suggest that no significant amount of electrons derived from the BES was harvested by the cells. Overall, this work points out that electro‐fermentation can be used to trigger metabolic pathways and improve product formation, even when the used microbe cannot be considered electroactive. Accordingly, further studies are required to unveil the underlying (regulatory) mechanisms.

Enhanced production of butanol and xylosaccharides from Eucalyptus grandis wood using steam explosion in a semi-continuous pre-pilot reactor

In this work, the co-production of xylosaccharides and butanol from Eucalyptus grandis wood using steam explosion pretreatment in a biorefinery platform was investigated. The effect of different temperature conditions on xylosaccharide production and selectivity as well as enzymatic hydrolysis of pretreated E. grandis substrates was studied using a pre-pilot steam explosion reactor. Under selected conditions (200 °C, 1.5 MPa, 10 min), steam explosion pretreatment led to about 50% xylan recovery as xylooligosaccharides (XOS) and xylose and 80% glucan enzymatic hydrolysis even at high solid loading. A xylosaccharide-rich hydrolysate was effectively separated by ion-exchange and resin treatment, containing mostly xylose and xylobiose as XOS. The fermentation performance of four native butanol-producing strains (Clostridium acetobutylicum DSM 1731, Clostridium beijerinckii DSM 6422, Clostridium beijerinckii DSM 6423 and Clostridium saccaroperbutylacetonicum DSM 14923) were evaluated using semi-synthetic media and enzymatic hydrolysates for butanol production. Results showed that ABE strain Clostridium beijerinckii DSM 6422 and IBE strain Clostridium beijerinckii DSM 6423 are robust native strains for butanol production from lignocellulosic materials. This work highlights a potential integrated biorefinery process for the efficient utilization of E. grandis wood to produce biofuels and biochemicals.