Project description:This study explores the production of furfural, xylose and methylxylosides through the methanolysis of wood flour using anhydrous HCl gas. The process involves methanolysis of wood flour with HCl gas under pressure to generate methylxylosides, which are subsequently converted to xylose and furfural via autohydrolysis in a Parr batch reactor system. The methanolysis was conducted in temperature-controlled HCl gas reactor employing 24 h reaction time and 50 % methanol content in wood flour. During the methanolysis step with HCl gas, 65 % of the available xylan in wood flour was converted to water-soluble methylxylosides, xylose, xylooligosaccharides (XO) and water-soluble methyl xylooligosaccharides (MXO). Methanolysis filtrates were then autohydrolyzed with Parr 50 mL batch reactor system to xylose and furfural in two different pH values at 180 °C. The highest furfural yield of 91 % from methanolysis filtrate was achieved with pH 1.2 and 25 min reaction time.
Project description:In the context of a biorefinery, lignocellulosic materials represent an important source of raw material for the bioconversion of cellulose, hemicellulose, and lignin into value-added products, such as xylose for fermentation, oligosaccharides, and bioplastics for packaging. Among the most abundant lignocellulosic materials in Brazil, sugarcane bagasse biomass stands out, as it is rich in cellulose and hemicellulose. In this context, through an experimental design, this study developed a robust enzyme cocktail containing xylanases and accessory enzymes to complete the hydrolysis of xylan from sugarcane bagasse, obtaining a low xylose yield and concentration (9% and 1.8 g/L, respectively, observed in experiment number 16 from the complete hydrolysis of a xylan assay), a fermentable sugar that is important in the production of second-generation ethanol, and a high xylooligosaccharides (XOS) yield and concentration (93.1% and 19.6 g/L, respectively, obtained from a xylooligosaccharides production assay); in general, xylan has prebiotic activities that favor an improvement in intestinal functions, with immunological and antimicrobial actions and other benefits to human health. In addition to completely hydrolyzing the sugarcane bagasse xylan, this enzymatic cocktail has great potential to be applied in other sources of lignocellulosic biomass for the conversion of xylan into xylose and XOS due to its enzymes content, involving both main chain and pendant groups hydrolysis of hemicelluloses.
Project description:BACKGROUND:Pretreatment is a critical step in the conversion of lignocellulose to fermentable sugars. Although many pretreatment processes are currently under investigation, none of them are entirely satisfactory in regard to effectiveness, cost, or environmental impact. The use of hydrogen peroxide at pH 11.5 (alkaline hydrogen peroxide (AHP)) was shown by Gould and coworkers to be an effective pretreatment of grass stovers and other plant materials in the context of animal nutrition and ethanol production. Our earlier experiments indicated that AHP performed well when compared against two other alkaline pretreatments. Here, we explored several key parameters to test the potential of AHP for further improvement relevant to lignocellulosic ethanol production. RESULTS:The effects of biomass loading, hydrogen peroxide loading, residence time, and pH control were tested in combination with subsequent digestion with a commercial enzyme preparation, optimized mixtures of four commercial enzymes, or optimized synthetic mixtures of pure enzymes. AHP pretreatment was performed at room temperature (23°C) and atmospheric pressure, and after AHP pretreatment the biomass was neutralized with HCl but not washed before enzyme digestion. Standard enzyme digestion conditions were 0.2% glucan loading, 15 mg protein/g glucan, and 48 h digestion at 50°C. Higher pretreatment biomass loadings (10% to 20%) gave higher monomeric glucose (Glc) and xylose (Xyl) yields than the 2% loading used in earlier studies. An H2O2 loading of 0.25 g/g biomass was almost as effective as 0.5 g/g, but 0.125 g/g was significantly less effective. Optimized mixtures of four commercial enzymes substantially increased post-AHP-pretreatment enzymatic hydrolysis yields at all H2O2 concentrations compared to any single commercial enzyme. At a pretreatment biomass loading of 10% and an H2O2 loading of 0.5 g/g biomass, an optimized commercial mixture at total protein loadings of 8 or 15 mg/g glucan gave monomeric Glc yields of 83% or 95%, respectively. Yields of Glc and Xyl after pretreatment at a low hydrogen peroxide loading (0.125 g H2O2/g biomass) could be improved by extending the pretreatment residence time to 48 h and readjusting the pH to 11.5 every 6 h during the pretreatment. A Glc yield of 77% was obtained using a pretreatment of 15% biomass loading, 0.125 g H2O2/g biomass, and 48 h with pH adjustment, followed by digestion with an optimized commercial enzyme mixture at an enzyme loading of 15 mg protein/g glucan. CONCLUSIONS:Alkaline peroxide is an effective pretreatment for corn stover. Particular advantages are the use of reagents with low environmental impact and avoidance of special reaction chambers. Reasonable yields of monomeric Glc can be obtained at an H2O2 concentration one-quarter of that used in previous AHP research. Additional improvements in the AHP process, such as peroxide stabilization, peroxide recycling, and improved pH control, could lead to further improvements in AHP pretreatment.
Project description:Purified endoxylanase from Thermomyces lanuginosus PC7S1T was immobilized in calcium alginate, resulting in a yield of 78.5% and a reusability for 11 cycles. The stability of the immobilized enzyme was given for a pH range of 4 to 9 for 96 h. Endoxylanase immobilized in calcium alginate at 65 °C exhibited thermal stability equal to the soluble enzyme for 5 h, and at high temperatures of 75 °C and 85 °C showed half-lives of 4 and 3 h, respectively. Both soluble endoxylanase and immobilized forms were able to hydrolyze hemicellulose, obtained from low-lignin sorghum biomass pretreated with 5% H2O2 and 2% NaOH, after 1 h of incubation at 65 °C, releasing a mixture of short-chain xylooligosaccharides (X2-X6). The highest amounts of XOS generated were those for X5 (24 to 40%), X4 (33 to 39%), and X3 (11 to 22%). These XOS acted as prebiotics, promoting the growth of the probiotic L. acidophilus, similar to glucose in the MRS broth. These results show the potential of low-lignin sorghum to generate XOS with prebiotic activity, suggesting the application of these compounds in the food industry.
Project description:Three lignocellulosic pretreatment techniques (ammonia fiber expansion, dilute acid and ionic liquid) are compared with respect to saccharification efficiency, particle size and biomass composition. In particular, the effects of switchgrass particle size (32-200) on each pretreatment regime are examined. Physical properties of untreated and pretreated samples are characterized using crystallinity, surface accessibility measurements and scanning electron microscopy (SEM) imaging. At every particle size tested, ionic liquid (IL) pretreatment results in greater cell wall disruption, reduced crystallinity, increased accessible surface area, and higher saccharification efficiencies compared with dilute acid and AFEX pretreatments. The advantages of using IL pretreatment are greatest at larger particle sizes (>75 µm).
Project description:Hydrogen chloride (HCl) gas is highly toxic to the human body. Therefore, HCl gas detection sensors should be installed at workplaces where trace HCl gas is continuously generated. Even though various polymer-based HCl-gas-sensing films have been developed, simpler and novel sensing platforms should be developed to ensure the cost effectiveness and reusability of the sensing platforms. Therefore, we present a simple strategy to fabricate reusable HCl-gas-sensing platforms using aminated polystyrene (a-PS) colloids and investigate their sensitivity, reusability, and selectivity using a quartz crystal microbalance (QCM). The reusable a-PS(1.0) colloidal sensor with a high degree of amination (DA) exhibited the highest binding capacity (102 μg/mg) based on the frequency change (Δf) during the HCl gas adsorption process. Further, its sensitivity and limit of detection (LOD) were 3.88 Hz/ppm and 5.002 ppm, respectively, at a low HCl gas concentration (<10 ppm). In addition, the sensitivity coefficient (k*) of the a-PS(1.0) colloid sensor with respect to HCHO was higher than that in the case of HF because of the lower binding affinity of the former with the a-PS(1.0) colloids. Based on these results, highly sensitive and reproducible a-PS colloids could be reused as an HCl-gas-sensing platform and used as an HCl sorbent in a gas column filter.
Project description:Xylooligosaccharides (XOS) have gained increased interest as prebiotics during the last years. XOS and arabinoxylooligosaccharides (AXOS) can be produced from major fractions of biomass including agricultural by-products and other low cost raw materials. Endo-xylanases are key enzymes for the production of (A)XOS from xylan. As the xylan structure is broadly diverse due to different substitutions, diverse endo-xylanases have evolved for its degradation. In this review structural and functional aspects are discussed, focusing on the potential applications of endo-xylanases in the production of differently substituted (A)XOS as emerging prebiotics, as well as their implication in the processing of the raw materials. Endo-xylanases are found in at least eight different glycoside hydrolase families (GH), and can either have a retaining or an inverting catalytic mechanism. To date, it is mainly retaining endo-xylanases that are used in applications to produce (A)XOS. Enzymes from these GH-families (mainly GH10 and GH11, and the more recently investigated GH30) are taken as prototypes to discuss substrate preferences and main products obtained. Finally, the need of new and accessory enzymes (new specificities from new families or sources) to increase the yield of different types of (A)XOS is discussed, along with in vitro tests of produced oligosaccharides and production of enzymes in GRAS organisms to facilitate use in functional food manufacturing.
Project description:As is well-known, endo-1,4-β-xylanase and β-xylosidase are the rate-limiting enzymes in the degradation of xylan (the major hemicellulosic component), main functions of which are cleavaging xylan to release xylooligosaccharides (XOS) and xylose that these two compounds have important application value in fuel, food, and other industries. This study focuses on enzymatic hydrolysis of poplar sawdust xylan for production of XOS and xylose by a GH11 endo-1,4-β-xylanase MxynB-8 and a GH39 β-xylosidase Xln-DT. MxynB-8 showed excellent ability to hydrolyze hemicellulose of broadleaf plants, such as poplar. Under optimized conditions (50°C, pH 6.0, dosage of 500 U/g, substrate concentration of 2 mg/mL), the final XOS yield was 85.5%, and the content of XOS2-3 reached 93.9% after 18 h. The enzymatic efficiency by MxynB-8 based on the poplar sawdust xylan in the raw material was 30.5%. Xln-DT showed excellent xylose/glucose/arabinose tolerance, which is applied as a candidate to apply in degradation of hemicellulose. In addition, the process and enzymatic mode of poplar sawdust xylan with MxynB-8 and Xln-DT were investigated. The results showed that the enzymatic hydrolysis yield of poplar sawdust xylan was improved by adding Xln-DT, and a xylose-rich hydrolysate could be obtained at high purity, with the xylose yield of 89.9%. The enzymatic hydrolysis yield was higher (32.2%) by using MxynB-8 and Xln-DT together. This study provides a deep understanding of double-enzyme synergetic enzymolysis of wood polysaccharides to valuable products.
Project description:BackgroundPopulus (poplar) tree species including hybrid varieties are considered as promising biomass feedstock for biofuels and biochemicals production due to their fast growing, short vegetative cycle, and widely distribution. In this work, poplar was pretreated with acetic acid (AC) to produce xylooligosaccharides (XOS), and hydrogen peroxide-acetic acid (HPAC) was used to remove residual lignin in AC-pretreated poplar for enzymatic hydrolysis. The aim of this work is to produce XOS and monosaccharides from poplar by a two-step pretreatment method.ResultsThe optimal conditions for the AC pretreatment were 170 °C, 5% AC, and 30 min, giving a XOS yield of 55.8%. The optimal HPAC pretreatment conditions were 60 °C, 2 h, and 80% HPAC, resulting in 92.7% delignification and 87.8% cellulose retention in the AC-pretreated poplar. The two step-treated poplar presented 86.6% glucose yield and 89.0% xylose yield by enzymatic hydrolysis with a cellulases loading of 7.2 m/g dry mass. Very high glucose (93.8%) and xylose (94.6%) yields were obtained with 14.3 mg cellulases/g dry mass. Both Tween 80 and β-glucosidase enhanced glucose yield of HPAC-pretreated poplar by alleviating the accumulation of cellobiose. Under the optimal conditions, 6.9 g XOS, 40.3 g glucose, and 8.9 g xylose were produced from 100 g poplar.ConclusionsThe AC and HPAC pretreatment of poplar represented an efficient strategy to produce XOS and fermentable sugars with high yields. This two-step pretreatment was a recyclable benign and advantageous scheme for biorefinery of the poplar into XOS and monosaccharides.