Total Reducing Sugar Yield Research Articles

Utilization of banana peel waste for the production of bioethanol and other high-value-added compounds

Bananas (Musa sp.) are one of the most produced crops worldwide, surpassing 100 million tons per year, and considering that approximately 30 % of their weight consists of peels, the annual generation of these residues is significant. Banana residues are rich in nutrients; nowadays, they are all disposed of in landfills. As a novelty, this work showed that it could be used for other things, such as alcohol, D-Limonene, and citric acid production. This study characterized the biomass, optimized pretreatment, and enzymatic hydrolysis and evaluated the fermentation process using this biomass, using the central composite design (CCD) methodology. The planning variables included sulfuric acid (H2SO4) concentration, banana peel waste mass in the pretreatment stage, and commercial cellulase enzyme (Sigma-Aldrich) concentration in the enzymatic hydrolysis stage. The responses quantified were sugars, ethanol, and acids using high-performance liquid chromatography, and the D-Limonene content was determined on a gas chromatograph coupled to a mass spectrometer. The highest yield of total reducing sugars found after enzymatic hydrolysis was 11.88 g/L, while the ethanol yield was 1.37 g/L. In the recovery of D-Limonene, the obtained value was 0.56 mg/g. The values of fermentable sugars confirm banana peel waste as a potential biomass for biorefinery processes. Furthermore, the possibility of an integrated D-Limonene recovery is essential in obtaining bioproducts. In this sense, it can be concluded that utilizing these residues is promising in reducing environmental issues and valorizing food waste.

Ultrasound-assisted hydrolysis of food waste using glucoamylase: Statistical optimization and mechanistic analysis with molecular simulations

This paper reports investigations in food waste hydrolysis using ternary approach that combines statistical optimization, ultrasound-assisted enhancement of hydrolysis kinetics, and molecular simulations that provide physical insight into the process. Initial optimization of hydrolysis parameters (Box–Behnken design) resulted in total reducing sugar yield of 263.4 mg/g biomass in 42 h. Sonication of hydrolysis mixture at 35 kHz at 20 % duty cycle yielded 4× reduction in hydrolysis time with 22 % enhancement in TRS yield (320 mg/g biomass). Analysis of GLCM's secondary structure through FTIR spectra deconvolution revealed significant changes induced by sonication. Sonication led to reduction in α-helix, and increase in random coil content. Molecular dynamics simulations unveiled majority of amino acid residues associated with GLCM binding pocket in α-helix and random coil regions. Consequently, sonication widened the binding pockets, facilitating easier transport of substrate and product. This effect translated into faster kinetics of enzymatic food waste hydrolysis.

Chromium-based metal-organic framework, MIL-101 (Cr), assisted hydrothermal pretreatment of teff (Eragrostis tef) straw biomass

Teff (Eragrostis tef) is a staple crop and holds the biggest share of grains cultivated area in Ethiopia, consequently, a large quantity of Teff straw is produced. The Teff straw was pretreated for the first time with Chromium-based Metal-Organic Framework, MIL-101(Cr), assisted hydrothermal method at temperatures ranging from 160 to 240 °C for 1/2, 1, or 2 h time independently. With an increase of pretreatment severity, the yield of total reducing sugar (TRS) was increased until reaching maximum (185 mg g−1). The identified optimum hydrothermal pretreatment condition, (180 °C and 1 h), had a feature of higher TRS yield and lower furfural concentration. The morphological analysis showed that treated Teff straw had degraded structure, higher surface area, and distorted bundles than native Teff straws. This study insight into MOFs’ application in lignocellulose biomass processing, and optimizing the pretreatment condition of Teff straw biomass.

A sustainable synthesis of polyhydroxyalkanoate from stubble waste as a carbon source using Pseudomonas putida MTCC 2475.

Polyhydroxyalkanoates (PHAs) are biodegradable polymers that can be produced from lignocellulosic biomass by microorganisms. Cheap and readily available raw material, such as corn stover waste, has the potential to lessen the cost of PHA synthesis. In this research study, corn stover is pretreated with NaOH under conditions optimized for high cellulose and low lignin with central composite design (CCD) followed by characterization using Fourier-transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), and scanning electron microscopy (SEM). Design expert software performed further optimization of alkali pretreated corn stover for high total reducing sugar (TRS) enhancement using CCD using response surface methodology (RSM). The optimized condition by RSM produced a TRS yield of 707.19mg/g. Fermentation using corn stover hydrolysate by Pseudomonas putida MTCC 2475 gave mcl-PHA detected through gas c hromatography - t andem m ass s pectrometry (GC-MS/MS) and characterization of the PHA film by differential scanning calorimetry (DSC), FTIR, and nuclear magnetic resonance (NMR). Thus, this research paper focuses on using agriculture (stubble) waste as an alternative feedstock for PHA production.

H-ZSM-5/GO Composites as a Catalyst for the Hydrolysis of Cellulose

Abstract: H-ZSM-5/GO composites were prepared for the catalytic hydrolysis of cellulose in ionic liquids of 1-butyl-3-methylimidazole chloride salt to obtain sugars. The materials catalyzed the hydrolysis of cellulose to produce total reducing sugar (TRS), super to ZSM-5 or graphene oxide (GO). The results suggested that the acidic sites of both materials and the mesopores of the composites enhanced the catalytic performance. With the optimized reaction conditions (e.g., ratio of catalyst to cellulose, temperature, reaction time), 87.8% yield of TRS was obtained.

Raspberry-like hydroxyl-rich carbon layer coated solid catalyst to promote the hydrolysis of cellulose to sugar through local strong absorption

Raspberry-like carbon layer coated silica nanoparticles (MSN-x-NH2 @C) with controllable hydroxyl-rich rough surface is designed and synthesized by use of alkaline etching strategy and hydrothermal carbonization (HTC) method, which is utilized as catalyst to provide particular interaction with cellulose between the solid-solid interfaces. Through the control of surface roughness of the raspberry-like catalyst, it induces the formation of local area with strong absorption on the surface and thus enhance the capability to disrupt the aggregated structure of cellulose. Accordingly, in low acidic aqueous system with MSN-x-NH2 @C, the cellulose conversion rate is up to 91.99% with the total reducing sugar yield at 81.46% and glucose yield of ∼70% after the second hydrolysis in aqueous system. Furthermore, the obtained MSN-x-NH2 @C exhibits good thermal stability and reusability. This research provides a new perspective for the design and preparation of nano-sized solid catalyst with controlled surface topological structure to enable the surface adsorption capability towards cellulose more efficiently in aqueous system.

Enzymolysis kinetics of corn straw by impeded Michaelis model and Box-Behnken design

Enzymatic hydrolysis is an essential step in the lignocellulosic biorefining process. In this paper, Box-Behnken was used to optimize the enzymatic hydrolysis process of corn stalk, and the promotion effect of three typical surfactants on the enzymatic hydrolysis process was investigated. The experimental results showed that the total reducing sugar yield reached 67.6% under the best-predicted conditions. When the concentration of Tween 80 is 0.1%, it could be increased to 80.2%. In addition, the Impeded Michaels Model (IMM) is introduced in this study to describe the enzymatic hydrolysis process of corn stalks. Finally, the initial contact coefficient between the enzyme and cellulose (Kobs,0) and the gradual loss coefficient of enzyme activity (ki) caused by reaction obstruction were obtained by fitting data, which successfully verified the rationality of the model.

Designing of recombinant hydrolytic enzymes cocktail for effective saccharification of delignified rice straw

Enzymatic saccharification is the most crucial and challenging step in lignocellulosic biorefineries for bioethanol production. The enzymatic saccharification requires a cocktail of hydrolytic enzymes, cellulases and hemicellulases for effective degradation of biomass. However, the complete bioconversion of biomass is governed by the proportion and synergistic action of hydrolytic enzymes present in the cocktail. Thus, this study aims to formulate an enzyme cocktail consisting of recombinant cellulases (bifunctional cellulolytic chimeric enzyme, CtGH1-L1-CtGH5-F194A and cellobiohydrolase, CtCBH5A) and xylanases (endo-1,4-β-xylanase, CtGH11A, and β-xylosidase, BoGH43), all in crude form (unpurified enzyme present in the total cellular proteins) for producing high yield of total reducing sugars using choline chloride:acetic acid pretreated rice-straw (CApRS). D-optimal mixture design approach was used for statistically designing the optimal proportion of crude recombinant enzyme cocktail for hydrolysis of delignified rice straw. The developed enzyme cocktail comprising chimera, CtCBH5A, CtGH11 and BoGH43 in ratio 35.1:41.8:10.0:13.1 resulted in cellulolytic enzyme activity, 5.6 FPU/mL and xylanase activity, 102 U/mL. Enzymatic saccharification of 3% (w/v) delignified CApRS using developed enzyme cocktail at 187 FPU/gCApRS, pH 5.8 and 35 °C, yielded total reducing sugar of 498 mg/gbiomass, glucose yield (410 mg/g) and xylose yield (71.3 mg/g) resulting in 72% saccharification efficiency. Whereas, significantly low total reducing sugar yield of 80 mg/g and 218 mg/g were released from raw RS and partially pretreated RS, respectively, by the developed cocktail under the same hydrolytic conditions, thereby inferring the effectiveness of the enzyme cocktail against CApRS biomass. The developed enzyme cocktail was compared with a commercial enzyme cocktail and prospects its applicability in lignocellulosic biorefineries.

Enzymatic digestibility of lignocellulosic wood biomass: Effect of enzyme treatment in supercritical carbon dioxide and biomass pretreatment

Energy and resource intensive mechanical and chemical pretreatment along with the use of hazardous chemicals are major bottlenecks in widespread lignocellulosic biomass utilization. Herein, the study investigated different pretreatment methods on spruce wood namely supercritical CO2 (scCO2) pretreatment, ultrasound-assisted alkaline pretreatment, and acetosolv pulping-alkaline hydrogen peroxide bleaching, to enhance the enzymatic digestibility of wood using optimized enzyme cocktail. Also, the effect of scCO2 pretreatment on enzyme cocktail was investigated after optimizing the concentration and temperature of cellulolytic enzymes. The impact of scCO2 and ultrasound-assisted alkaline pretreatments of wood were insignificant for the enzymatic digestibility, and acetosolv pulping-alkaline hydrogen peroxide bleaching was the most effective pretreatment that showed the release of total reducing sugar yield (TRS) of ∼95.0 wt% of total hydrolyzable sugars (THS) in enzymatic hydrolysis. The optimized enzyme cocktail showed higher yield than individual enzymes with degree of synergism 1.34 among the enzymes, and scCO2 pretreatment of cocktail for 0.5–1.0 h at 10.0–22.0 MPa and 38.0–54.0 °C had insignificant effect on the enzyme's primary and global secondary structure of cocktail and its activity.

Continuous and effective pretreatment of sugarcane bagasse lignocellulose with DBU/glycerol to enhance enzymatic saccharification

The new deep eutectic solvent (DES) synthesized from 1,8-Diazabicyclo [5.4.0] undec-7-ene (DBU) and glycerol was utilized for the continuous pretreatment of sugarcane bagasse (SCB) and facilitating enzymatic saccharification. Under the optimal pretreatment conditions (DBU/glycerol weight ratio of 1:1.7, 160 ℃, 70 min) and enzymatic hydrolysis of the pretreated SCB by Cellulases Cellic CTec 2 (20 FPU per gram of substrate) at 50 ℃ for 72 h, the total reducing sugar content (TRSC) and total reducing sugar yield (TRSY) were 15.76 g/L and 0.51 g/g, respectively, and the lignin removal rate was 83.21%. After five recycles, the recovered pretreatment liquid maintained excellent pretreatment performance, where TRSC (14.68 g/L) and TRSY (0.48 g/g) can remain at high levels. Besides, the physicochemical properties of the DBU/glycerol solvent system were characterized, and the structural changes of SCB after pretreatment to enhance enzymatic hydrolysis were investigated by field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), and Fourier transform infrared analyzer (FT-IR). The results show that DBU/glycerol pretreatment is a green, efficient, and sustainable lignocellulosic pretreatment strategy.

Preparation and Catalytic Properties of Graphene Oxide/ Phosphotungstic Acid Composites

Background: Cellulose structures are in stable crystallineform. The hydrolysis of cellulose to small reducing sugars is difficult, but essential for its utilization Objective: To investigate the effect of graphene oxide (GO) loading on the catalytic performance of phosphotungstic acid (HPW) for the catalyzed hydrolysis of cellulose, with the purpose to get high yield of total reducing sugar (TRS). Methods: Graphene oxide/phosphotungstic acid (GO/HPW) composites were prepared using a liquid-phase composite method. The materials were applied to catalyze hydrolysis of microcrystalline cellulose in 1-butyl-3-methylimidazole chloride ionic liquid ([Bmim]Cl). The samples were characterized by Powder X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), Field emission scanning electron micrographs (FE-SEM), pyridine IR and acid-base chemical titration. Results: The Brønsted acidic sites were the main source of acidity in the composites and its concentration was determined to be 0.96 mmol/g. With the use of the GO/HPW composite as catalysts for cellulose hydrolysis, TRS yield of 90.5 % was obtained. Conclusion: GO/HPW composites retained the functional groups of both materials. It was the Brønsted acidic sites in the materials that effectively promoted the cellulose hydrolysis reaction. The structures of GO/HPW with the agglomeration of HPW scattered on GO had high accessibility of acidic sites and fast mass transfer of the reducing sugars to the outside of the catalysts in time to prevent their further conversion into by-products. TRS yield of 90.5 % was obtained from the hydrolysis of cellulose catalyzed by the GO/HPW (1:1.5) composites at 115 ℃ for 4 h using catalysts to cellulose 1:1 ratio.

Hydroxy sulfonic acid catalyzed hydrolysis of cellulose

Development of efficient catalytic methods for the hydrolysis of cellulose is a major research challenge in sustainable biofuel and polymer areas. In this study five hydroxy sulfonic acids were studied as simple model compounds for cellulase enzyme for the hydrolysis of cellulose. The catalytic activities were measured by analysis of total reducing sugar (TRS) yields produced in a series of reactions carried out at 150–190 °C using 0.050 M aqueous hydroxy sulfonic acid solutions. The highest catalytic activity was observed with isethionic acid, producing 62.7% TRS yield at 180 °C after 4 hr. In the second phase of the work, Density Functional Theory (DFT) calculations were used to study the interactions between hydroxy sulfonic acids and cellulose model compound D-cellobiose to supplement the experimental results. The D-cellobiose – hydroxy sulfonic binding energies and the distance between glycosidic oxygen and -SO3H acidic H were evaluated and the -SO3H to glycosidic oxygen distance was identified as the more important parameter co-related to the catalytic activity. The isethionic acid with highest cellulose hydrolysis activity showed the shortest -SO3H to glycosidic oxygen distance of 1.744 Å.

Using citric acid to suppress lignin repolymerization in the organosolv pretreatment of corn stalk

The esterification reactions that occur between citric acid (CA) and lignin on hydroxyl groups of side chains can inhibit the formation of reactive intermediates in acid-catalyzed pretreatments. The effects of CA on dilute hydrochloric acid (1.0%) pretreatments of corn stalk in hydrated 1,4–dioxane were investigated with sodium hypophosphite (0.5%) as the catalyst at 95 °C for 3.5 h. The resulting cellulose recovery (62.5–80.9%) and lignin removal (72.5–85.0%) after adding CA were significantly higher than those in the pretreatment without CA. Water content influenced pretreatment and delignification when CA was added into the treatment systems. The highest yield of total reducing sugar (97.1%) was achieved during the enzymatic hydrolysis of cellulose-rich materials obtained from the pretreatment with 11.4% water content. Meanwhile, β–O–4′ in the extracted lignin increased from 22.4/100Ar to 41.2/100Ar with increasing water content from 11.4% to 25.9%. These findings implied that CA as a stabilization agent could inhibit the repolymerization of degraded lignin fragments due to the esterification between CA and lignin and then improved the enzymatic saccharification of cellulose.

Simultaneous saccharification of hemicellulose and cellulose of corncob in a one-pot system using catalysis of carbon based solid acid from lignosulfonate.

The drive towards sustainable chemistry has inspired the development of active solid acids as catalysts and ionic liquids as solvents for an efficient release of sugars from lignocellulosic biomass for future biorefinery practices. Carbon-based solid acid (SI-C-S-H2O2) prepared from sodium lignosulfonate, a waste of the paper industry, was used with water or ionic liquid to hydrolyze corncob in this study. The effects of various reaction parameters were investigated in different solvent systems. The highest xylose yield of 83.4% and hemicellulose removal rate of 90.6% were obtained in an aqueous system at 130 °C for 14 h. After the pretreatment, cellulase was used for the hydrolysis of residue and the enzymatic digestibility of 92.6% was obtained. Following these two hydrolysis steps in the aqueous systems, the highest yield of total reducing sugar (TRS) was obtained at 88.1%. Further, one-step depolymerization and saccharification of corncob hemicellulose and cellulose to reducing sugars in an IL-water system catalyzed by SI-C-S-H2O2 was conducted at 130 °C for 10 h, with a high TRS yield of 75.1% obtained directly. After recycling five times, the solid acid catalyst still showed a high catalytic activity for sugar yield in different systems, providing a green and effective method for lignocellulose degradation.

Two-stage acid-alkali pretreatment of vetiver grass to enhance the subsequent sugar release by cellulase digestion

Vetiver grass (VG; Chrysopogon zizanioides) can be used as a potential feedstock for biofuels and biochemical production due to its chemical composition, which is rich in cellulose, hemicellulose, and lignin. However, the complex and recalcitrant nature of a lignocellulosic biomass requires a pretreatment process to disrupt the lignin seal in the cell wall in order to increase the accessibility for enzymatic hydrolysis and to liberate the sugars contained within the cellulose. The sequential acid-base pretreatment of VG using dilute sulphuric acid (H2SO4) followed by sodium hydroxide (NaOH) was found to be optimal when using a 5% (w/v) VG loading in 0.5% (v/v) H2SO4 at 120 °C, 60 min in the first stage followed by 4% (w/v) NaOH at 120 °C, 60 min in the second stage. This gave the highest yield of total reducing sugars (32.6 g/L as 22.0 and 10.6 g/L of hexose and pentose, respectively) in the subsequent cellulase digestion, which was 10.3% and 46.1% higher than that obtained with the single-stage alkali (24.1 g/L) or acid (13.5 g/L) pretreatment, respectively.

Enhanced enzymatic digestibility of water lettuce by liquid hot water pretreatment

Liquid hot water (LHW) pretreatment conditions were optimized from various severities using response surface methodology to convert water lettuce (WL; Pistia stratiotes) to fermentable sugars to enhance the enzymatic digestibility of pretreated WL. The LHW pretreatment was performed using a 2% (w/v) WL loading over a temperature range of 152–208 °C and pretreatment times of 9–51 min. The optimal conditions for the LHW pretreatment were determined to be 190.7 °C for 51 min [a severity factor (log R0) of 4.38], which gave the highest total reducing sugar yield (17.26 g/L) and a high removal level of hemicellulose (68.32%) in the subsequent cellulase digestion. The physical structure of WL after pretreatment revealed changes to the WL structure that increased its susceptibility to attack by enzymes; an increased crystallinity, surface area, and total pore volume together with a lower hemicellulose and lignin content compared to the untreated WL.

SUSTAINABLE ENERGY-EFFICIENT CONVERSION OF WASTE TEA LEAVES TO REDUCING SUGAR: OPTIMIZATION AND LIFE-CYCLE ENVIRONMENTAL IMPACT ASSESSMENT

Innovative protocols involving energy-proficient pretreatment of waste tea leaves (WTL) for preparation of cellulose and its subsequent photocatalytic hydrolysis (PH) for production of total reducing sugar (TRS) have been reported. The WTL was subjected to alkali pretreatment (60 °C, 1 h) followed by bleaching (employing peracetic acid, 65 °C, 2 h) in a quartz halogen irradiated batch reactor (QHIBR) for efficient separations of lignin and hemicellulose fractions to produce WTL derived cellulose fiber (WTLDCF; 94.5% cellulose). Consequent PH of WTLDCF in QHIBR using combination of Amberlyst-15 and nano-TiO2 catalysts was optimized (parameters: 40 min, 70 °C, 1:30 WTLDCF to water weight ratio and 5 wt. % catalyst concentration) employing Taguchi design that provided maximum 68.25% TRS yield. The QHIBR demonstrated faster hydrolysis and superior energy-efficiency over conventional reactor owing to quartz halogen irradiation. Life cycle assessment indicated an acceptable global warming potential of 2.215 kg CO2 equivalent; thus, establishing an energy-efficient environmentally sustainable WTL valorization process.

Pretreatment of Corn Stover by Levulinic Acid-Based Protic Ionic Liquids for Enhanced Enzymatic Hydrolysis

Economically competitive and sustainable dissolution pretreatment technology is in high demand due to the remarkable achievement in the pretreatment of lignocellulose using ionic liquids (ILs) for enhanced enzymatic hydrolysis. Herein, levulinic acid (Lev), a well-known bio-based platform chemical derived from carbohydrates, in conjunction with a series of organic superbases, was used as a raw chemical to prepare a new class of protic ILs (PILs). The findings indicated that the obtained PILs exhibited a good solubility of up to 10 wt % toward corn stover-based lignocellulose at 140 °C in 40 mins, based on which a facile and efficient dissolution pretreatment technology was developed for enhanced enzymatic hydrolysis of corn stover. The facile dissolution and regeneration of corn stover resulted in significant changes in the composition and physical–chemical structures. These changes led to significantly enhanced enzymatic hydrolysis behavior of the pretreated sample with total reducing sugar and glucose yields of 0.8 and 0.49 g/g within 48 h under optimal conditions, respectively. The Kamlet–Taft solvent parameters of the PILs were systematically evaluated, and the interaction between the lignocellulose and PILs was studied by 13C NMR, indicating that the satisfactory performance of PILs was benefited from the higher ß parameter value and the stronger interactions, particularly the existence of the extra hydrogen-bond interaction from the potential keto–enol tautomerism of the ketone groups in levulinate anions. The compositional and physicochemical changes in the lignocellulose were systematically evaluated to achieve an in-depth understanding of the dissolution activation mechanism using various characterization techniques. The findings also showed that 44.3% of lignin could be fractionated during the facile dissolution and regeneration process, together with a transformation of the crystalline structure of cellulose I to cellulose II and of the packed morphology to a porous one. The structure of the fractionated lignin was also characterized for a better understanding of the fractionation process. The study demonstrated a new sustainable dissolution pretreatment technology for enhanced enzymatic hydrolysis, which can provide significant insights into the design of new solvents for biomass dissolution and processing.

Acid Leaching Vermiculite: A Multi-Functional Solid Catalyst with a Strongly Electrostatic Field and Brönsted Acid for Depolymerization of Cellulose in Water.

Vermiculite is a natural mineral. In this study, vermiculite and acid-activated vermiculite was used as a solid acid catalyst for the hydrolysis of cellulose in water. The catalysts were characterized by XRD, FT-IR, and BET. The effects of time, temperature, mass ratio and water amount on the reaction were investigated in the batch reactor. The results showed that the highest total reducing sugars (TRS) yield of 40.1% could be obtained on the vermiculite activated by 35 (wt)% H2SO4 with the mass ratio of catalyst to cellulose of 0.18 and water to cellulose of 16 at 478 K for 3.5 h. The acid-activated vermiculite was a stable catalyst through calcination at 628 K and the yield of TRS decreased to 36.2% after three times reuse. The results showed that the crystal structure of vermiculite was destroyed and the surface -OH groups increased after the acid treatment. However, the synergistic effect of a strongly electrostatic polarization and Brönsted acid was responsible for the efficient conversion of cellulose. The mechanism of cellulose hydrolysis on the acid-activated vermiculite was suggested. This work provides a promising strategy to design an efficient solid catalyst for the cellulose hydrolysis, and expands the use of vermiculite in a new field.

Construction of an enzymatic shuttling compartment based on reverse micellar for bamboo biomass hydrolysis in ionic liquids

The enzymatic saccharification of regenerated lignocellulose must occur separately due to the toxicity of ionic liquids to cellulase. Therefore, it is important to develop a biocompatible IL-cellulase system which effectively achieves activation and saccharification of lignocellulose. For this purpose, a dual-phase “enzyme-shuttling compartment” was constructed in this study. Tween 80 was found to form reverse micelles in the isooctane-IL two-liquid phase, acting as a microenvironment that maintains the energetic conformation of the reactive cellulase. The activated bamboo biomass was enzymatically hydrolyzed in 20% (w/v) 1-ethyl-3-methylimidazolium dimethyl phosphate and 50 mM citrate buffer at 50 °C, achieving a high total reducing sugar yield of 71.2% and maintaining an enzymatic activity of 91.2% after 24 h. Thus, an efficient system with the simultaneous activation and saccharification of natural biomass was successfully developed in a one-pot procedure at low temperatures, ensuring large-scale biomass conversion into biofuels and biological products.