Filter Paper Units Research Articles

Obtaining oil and fermentable sugars from olive pomace using sequential supercritical fluid extraction and enzymatic hydrolysis

Olive pomace has high economic potential and the supercritical technology for extraction produces oil with potential use in the food and biofuels industries. The remaining solid can be used to obtain fermentable sugars from the hydrolysis. The present work aims to evaluate the effect of supercritical fluid extraction (SFE) of pomace olive oil at 60 and 40 °C and 18 and 22 MPa on oil yield and composition. Furthermore, use the remaining solid of SFE to obtain fermentable sugars from enzymatic hydrolysis. Kinetic curves were obtained for SFE, with the best oil yields 19.36 ± 2.43 wt% obtained at 40 °C / 22 MPa. The antioxidant activity was better for (60 °C / 22 MPa) presenting 137.87 ± 1.04 µmol TEAC / g am. dry. The highest yield of fermentable sugar (YFS) was 41.02 ± 4.79 g defatted olive pomace (DOP) for 26 Filter Paper Unit (FPU) / 1% solid loading (CS).

Culture of Trichoderma sp. with biochar to produce high-activity cellulase in a laboratory

Biochar (BC) was used in Trichoderma sp. culture to produce high-activity cellulase on a laboratory scale. The biochar was added into the flask before being applied to the fermenter to identify the enhancement effect and to determine the best amount of addition and the most suitable incubation period. Cellulase production was performed with a working volume of 4 L, and enzymatic hydrolysis was conducted to evaluate the saccharification ability of the enzyme. During incubation, the activities of three enzymes (Endoglucanase (EG), β-glucosidase (BGL), and cellobiohydrolase (CBH)) were measured for three days, and the cellulase activity was determined using a filter paper unit (FPU). In flask scale, EG, BGL, and CBH activities were increased by 1.4, 2.1, and 1.8 folds, respectively, and the incubation period was shortened by adding BC. In the fermenter scale, EG, BGL, and CBH activities were noticeably enhanced by 12.1, 5.8, and 7.2 folds, respectively, and FPU was 42.1 (9.8 folds). Additionally, the conversion rates of cellulose and steam exploded softwood and hardwood were 109.4%, 75.4%, and 87.3%, which were similar to a commercial enzyme (Cellic CTecⅡ). This study demonstrated that biochar could be used to produce high-activity cellulase in a shorter period and suggests a novel method for effective cellulase production.

Integrated rheological, morphological and biochemical approaches to investigate performances of an enzymatic secretome from Talaromyces sp. BRG1 to degrade pretreated sugarcane bagasse and paper-pulp

Exploring cellulolytic secretome is a promising strategy to produce a cost-effective enzyme preparation for biorefinery of lignocellulosic waste. In this study, a fungal secretome from Talaromyces sp. BRG1 was produced with pretreated sugarcane bagasse (SCB) as the inductive substrate. Hydrolysis capacity of BRG1 secretome and commercial enzyme Cellic® Ctec2 were compared on SCB and then applied to paper pulp (PP). Normalised activity profile of cocktails based on filter paper unit activity (FPU) indicated that BRG1 secretome exhibited a higher endo-glucanase (CMCU, ×4.4) and endo-xylanase activities (EnXU, ×1.6) but lower avicelase (AVCU, /8.4) and cellobiase (CBU, /3.4) activities. Lignocellulolytic cocktails were examined on SCB as reference before applied on PP at 3% w/v solid loading in batch mode for 24 h. For both substrates and enzyme loading ratio (0.3 and 3.0 FPU/g cellulose), the glucose yield by BRG1 secretome was 2 times higher than Cellic® Ctec2. In-situ viscometry confirmed the quicker liquefaction efficiency of secretome over Ctec2. At 0.3 FPU/g cellulose, the suspension viscosity of PP collapsed completely after 12 h with BRG1 secretome whereas a limited reduction to 44% was observed with Ctec2. Ex-situ particle size distribution revealed a stronger reduction of coarse and fine particles with BRG1 secretome compared to Ctec2.

Valorization of spent coffee grounds through integrated bioprocess of fermentable sugars, volatile fatty acids, yeast-based single-cell protein and biofuels production

Recovering nutrients from waste for biological processes aligns with sustainability principles. This study aimed to convert spent coffee grounds (SCG) into valuable products, including fermentable sugars, volatile fatty acids (VFAs), yeast-based single-cell protein and biofuels. Alkaline pretreatment was conducted before enzymatic hydrolysis, in which the pretreated SCG was hydrolyzed with varying enzyme loadings (20–60 filter paper units (FPU)/g-solid) and solid loadings (3–15 % w/v). The hydrolyzed slurry was utilized for VFAs and hydrogen production, yielding high values of 0.66 g/g-volatile solids (VS) and 109 mL/g-VS, respectively, using an enzyme loading of 50 FPU/g-solid and a solid loading of 3 % (w/v). The derived VFAs were used to cultivate a newly isolated yeast, Candida maltosa KKU-ARY2, resulting in an accumulated protein content of 43.7 % and a biomass concentration of 4.6 g/L. This study highlights the conversion of SCG into essential components, emphasizing the benefits of waste utilization through cascade bioprocesses.

Low-dosage enzymatic hydrolysis of organic municipal waste for sugar and ethanol production

AbstractEnzymatic hydrolysis (EH) of the organic fraction of municipal solid waste (OFMSW) is challenging due to its complexity and heterogeneous character. This study investigated the effects of OFMSW compositional characteristics and pretreatment techniques on the efficiency of EH. The experiments involved varying enzymatic dosages and pretreatment methods, including heat pretreatment and anaerobic incubation of raw OFMSW. Remarkably, after thermal pretreatment, a saccharification yield exceeding 50% was achieved with only 1 filter paper unit (FPU) of Cellic® Ctec3 and 0.5 glucoamylase unit (AGU) of AMG® 300 L BrewQ per g of dry OFMSW. This resulted in the recovery of 239 g of sugar per kilogram of dry OFMSW. The study revealed that the decomposition of OFMSW by enzymes is limited after reaching about 50% of the sugar recovery yield, and high concentrations of inhibitors such as organic acids, alcohols, and potentially other organics and inorganics may contribute to this limitation. These compounds, however, serve as excellent preservatives, achieving contamination control during biological conversion. Sugar-rich hydrolysates were subjected to ethanol fermentation using Saccharomyces cerevisiae, leading to the conversion of most hexose sugars into ethanol, reaching 331 g of ethanol per kilogram of dry OFMSW, and indicating that the limitation of OFMSW saccharification was not due to product inhibition. These findings highlight the need for further investigation of the saccharification process of biowaste to overcome inhibitory effects and subsequently achieve higher efficiency of the following bioprocesses.

An integrated biorefinery process for mandarin peel waste elimination

Mandarin peel waste, a representative agricultural waste, has been dumped on land and marine environments worldwide, causing significant environmental damage. Thus, a solution is urgently required to reduce direct dumping and convert mandarin peel waste into high value-added products. In this study, to achieve a zero-waste concept, an integrated biorefinery effectively reduced the mass of mandarin peel waste through consecutive processes that included washing, autohydrolysis, enzymatic hydrolysis, and melt compounding. This suggests that integrated biorefinery produced 92.4% of arabinose and arabinooligosaccharide by autohydrolysis and 81.9% of glucose by enzymatic hydrolysis using 5 filter paper unit of cellulase. Overall, the enzymatically hydrolyzed residue replaced 10% of polylactic acid for biocomposite production, which reached approximately 84% of the tensile strength level compared to that of neat polylactic acid. Finally, a concise economic assessment presents the feasibility of an integrated biorefinery by converting value-added products from mandarin peel waste to overcome the environmental and economic problems associated with direct dumping.

Integrated Bioprocess for Cellulosic Ethanol Production from Wheat Straw: New Ternary Deep-Eutectic-Solvent Pretreatment, Enzymatic Saccharification, and Fermentation

Wheat straw (WS) is an excellent raw material for biofuel ethanol production. However, the recalcitrance of WS prevents its efficient utilization. In this study, a novel ternary deep eutectic solvent (DES) was developed for enhancing component separation and enzymatic saccharification of WS. Without any detoxification and sterilization, the DES-treated WS hydrolysate was successfully used to produce ethanol. Overall, this research evaluated the effect of ternary DES pretreatment on WS at various temperatures and adjusted the enzyme load, substrate concentration, and fermentation method of treated WS. The results suggested that the cellulose recovery of treated WS after DES pretreatment (120 °C, 1 h) was 94.73 ± 0.22%, while the removal of xylan and lignin reached 89.53 ± 0.36% and 80.05 ± 0.62%, respectively. Importantly, at enzyme loading of 11.4 filter paper unit (FPU)/g WS with 16% fermentation substrate concentration, 91.15 ± 1.07% of cellulose was hydrolyzed, and the glucose yield was 71.58 ± 1.34%. The maximum ethanol yield of DES-treated WS was 81.40 ± 0.01%.

Optimization of Pretreatment Conditions and Enzymatic Hydrolysis of Corn Cobs for Production of Microbial Lipids by Trichosporon oleaginosus

Microbial lipids produced from lignocellulosic biomass are sustainable alternative feedstock for biodiesel production. In this study, corn cobs were used as a carbon source for lipid production and growth of oleaginous yeast Trichosporon oleaginosus. Lignocellulosic biomass was subjected to alkali and acid pretreatment using sulfuric acid and sodium hydroxide under different temperatures, catalyst concentrations and treatment times. Pretreatment of corn cobs was followed by cellulase hydrolysis. Hydrolysis of alkali pretreated (2% NaOH at 50 °C for 6 h, 1% NaOH at 50 °C for 16 h, 2% NaOH at 121 °C for 1 h, 1% NaOH at 121 °C for 2 h) and acid pretreated (1% H2SO4 120 °C for 20 min, and 2% H2SO4 120 °C for 10 min) corn cobs resulted in more than 80% of the theoretical yield of glucose. The effect of substrate (5, 10, 15 and 20%, g g−1) and cellulase loading (15 and 30 Filter Paper Units per gram of glucan, FPU g−1) on fermentable sugar yield was also studied. The maximal glucose concentration of 81.64 g L−1 was obtained from alkali-pretreated corn cobs (2% NaOH at 50 °C for 6 h) at 20% substrate loading and 30 FPU of Cellic CTec2 g−1 of glucan. Enzymatic hydrolysates of pretreated biomasses and filtrates of lignocellulosic slurries obtained after pretreatment were used for growth and lipid synthesis by T. oleaginosus. The highest lipid concentration of 18.97 g L−1 was obtained on hydrolysate of alkali-pretreated corn cobs (with 1% NaOH at 50 °C for 16 h) using a 15% (g g−1) substrate loading and 15 FPU g−1 of cellulase loading. Significant lipid accumulation was also achieved using undetoxified filtrates of pretreated slurries as substrates. Results showed that pretreated corn cobs and undetoxified filtrates are suitable carbon sources for the growth and efficient accumulation of lipids in T. oleaginosus.

Effects of Colloid Milling and Hot-Water Pretreatment on Physical Properties and Enzymatic Digestibility of Oak Wood

A two-step process using colloid milling (CM) and hot water (HW) treatment was evaluated for its ability to improve xylose recovery and the enzymatic digestibility of oak wood. In the first step, CM pretreatment was applied at a milling (feeding) speed of 100 mL/min with four different milling times (3, 6, 9, and 12 h), and the enzymatic digestibility and physical properties of each substrate were measured. In the second-step, the HW pretreatment was applied to enhance the enzymatic digestibility and xylan recovery at various reaction severities (Log R0) from 2.07 to 4.43 using 12 h colloid-milled (CM-treated) oak wood. Compared with untreated oak wood, CM not only significantly disrupted the structure of oak wood but also increased its Brunauer–Emmett–Teller surface area (42-fold) and pore volume (28-fold). The crystallinity of two-step-treated oak wood was decreased to 34.8, while the enzymatic digestibility of 12 h CM-treated oak wood was increased to 58.1% at enzyme loading of 30 filter paper units (FPU)/g glucan for 96 h. After HW treatment of CM-treated oak wood at Log R0 = 3.83, 80.7% of xylan recovery yield and 91.1% of enzymatic digestibility (with 15 FPU/g glucan at 96 h) was obtained, which was 84.2% higher than the enzymatic digestibility of untreated oak wood (6.9%).

Steam Pretreatment of Rice Hulls to Release Fermentable Saccharides: An Approach to Improve Recovery of (Hemi)Cellulosic Sugars Through Multivariate Design

The conversion of rice hulls into fermentable saccharides was explored through steam pretreatment employing 2.5% SO2. The interaction between temperature and time was assessed by means of the response surface method to achieve optimum contents of C6-sugars in water-insoluble solids (WIS) and C5-sugars in the liquor. Pretreatment carried out at 218°C for 2.3min released liquor containing 55.4g/L of sugars (29.1g/L of xylose). In parallel, the WIS was subjected to enzymatic saccharification using different solid and enzyme loads via an experimental design: assays using 22.0% WIS and 20.0 filter paper units (FPU)/g led to 90.6g/L of glucose, corresponding to a yield of 86.4% and an overall yield of 72.4%. The data reported are the highest ever found for such raw material, making it attractive to compete with conventional lignocellulosic biomass.

Biorefinery potential of chemically enhanced primary treatment sewage sludge to representative value-added chemicals - A de novo angle for wastewater treatment

Chemically enhanced primary treatment (CEPT) is an emerging sewage treatment strategy due to its high efficiency and small land requirement. CEPT sludge can be easily dewatered and used for energy recovery through incineration. However, with large amount of reusable nutrients (40% organic carbon, 23% lipids, and 17% protein), the value of CEPT sludge may have been underestimated. In this study, the biorefinery potential of CEPT sludge has been proven via production of 28.9 g/L ethanol or 50.3 g/L lactic acid (LA) or 1.43 filter paper unit (FPU)/mL cellulase from 10 g of CEPT sludge experiment. Inhibition on cell growth and potential inhibitors from plasticizers, pharmaceuticals, and surfactants were determined. Nevertheless, production titer was not affected or performed even better than the non-inhibitors controls. CEPT sludge showed significant potential in biochemical conversion, and the related products may offer an opportunity to support wastewater treatment toward sustainability and carbon neutrality.

A Simplified Procedure for Cellulase Filter Paper Assay

A new procedures to minimize labor intensiveness and complexity that has long been recognized in cellulase filter paper activity measurement (FPAase) described by the international union of pure and applied chemistry (IUPAC) was developed. It follows the main idea of IUPAC finding at least two cellulase dilutions have optical densities slightly more and less than a reference optical density of an arbitrary fixed 2 mg absolute glucose amount after a red-ox color reaction and due to cellulase-filter paper hydrolysis. The yielding glucose amount difference as compared to this reference is expressed in terms of absorbance difference percent determined by 3,5-dinitrosalicylic acid (DNS), in case of cellulase is cellubiase rich. If not, an external supplemental portion should be added. The intersection of the line of these two cellulase dilutions with abscissa intersect a vertical at a hypothetical 0% absorbance difference percent corresponding to a critical cellulase dilution exactly release this fixed arbitrary 2 mg glucose amount value. The factor 0.37 of this critical cellulase dilution equals to its filter paper units expressed in FPU per ml. A cellulase Cellic Ctech2 from novozymes has been tested with this methodology giving a satisfactory results with IUPAC procedures.

Isolation of a cellulase hyperproducing mutant strain of Trichoderma reesei

Trichoderma reesei has been employed as a commercial cellulase producer; however, the cellulase obtained has low β-glucosidase activity. Lactose is a promising cellulase inducer because it is easy to be dissolved, but its induction efficiency is lower than that of solid inducers, such as biomass. We aimed to obtain a cellulase-producing strain that is compatible to on-site production, and developed a cellulase hyperproducing mutant, T. reesei T1281, through repeated random mutagenesis. Protein quantification using Bradford method showed that T1281 produced 35.4 g/L (119 g/L using Lowry method) protein in 212 h. Moreover, T1281 exhibited the highest filter paper unit (15.6 U/mL) and β-glucosidase (53.8 U/mL) activity on lactose among the strains developed using random mutagenesis. Enzymes produced by T1281 hydrolyzed biomass into glucose and xylose without cellobiose and xylobiose accumulation, indicating that T1281 can be a potential ideal strain for industrial cellulase production.

Boosting second-generation ethanol titers from green coconut fiber by using high-concentration polyethylene glycol

Efforts have been made to improve and modify lignocellulosic pretreatments, but this step corresponds to a high cost in second-generation ethanol production. Previous studies have shown that high-concentration polyethylene glycol (PEG) is a possible way to dispense chemical and physicochemical pretreatments and increase ethanol production from lignocellulosic material. However, no operational conditions of this strategy were optimized. The present study investigated the effects of operational conditions on simultaneous saccharification and fermentation (SSF) performance using high PEG concentrations and green coconut fiber (GCF) as a substrate. The high-concentration PEG (150 g.L−1) increased ethanol production compared to the PEG-free, 10, and 50 g.L−1 PEG conditions. Using 20 % (w.v−1) solid loading, the batch SSF cultivation with PEG 1500 and without supplementary nutrients reached ethanol production and ethanol yield equal to 22.21 g.L−1 and 64.0 %, respectively. The batch SSF cultivation with 30 % (w.v−1) solid loading presented mass transfer limitations. The addition of salts, yeast extract, or peptone improved ethanol production. Inhibition by ethanol on the Saccharomyces cerevisiae CAT-1 strain was indifferent to the presence of PEG 1500, while the addition of PEG ensured the cellulolytic activity reuse after three fermentation cycles. The fed-batch SSF with PEG facilitated the GCF liquefaction so that it was possible to operate with up to 30 % (w.v−1) solid loading without problems of viscosity and free water. Using 150 g.L−1 PEG 1500 and enzyme loading equal to 13.3 filter paper unit. g−1, the fed-batch SSF reached 35.1 g.L−1 ethanol at 48 h, representing an increase of 70 % compared to the fed-batch SSF without PEG.

Carbonate-oxygen pretreatment of waste wheat straw for enhancing enzymatic saccharification

Waste wheat straw (WWS), as the main solid waste from straw-handling yard in pulp mill, is often managed by combustion or sanitary landfill, representing an inefficient use and causing a great threat to the environment. Therefore, it is highly desirable to develop a facile and cost-effective method for the efficient utilization of WWS. In this work, Na2CO3-O2 pretreatment was conducted to WWS for enhancing the sugar recovery by enzymatic hydrolysis. The results indicated that Na2CO3-O2 pretreatment showed high delignification selectivity at relatively low temperature (110 °C) and total titratable alkali (TTA) charge (12 %, as Na2O), with the lignin removal and pretreatment yield of 42.0 % and 67.9 %, respectively. The low pH value of the spent liquor after Na2CO3-O2 pretreatment (∼ 8.8) prevented the severe alkaline degradation of carbohydrates. At a cellulase loading of 20 filter paper unit (FPU)/g-cellulose, the total sugar recovery based on oven dry matter of raw material was up to 66.1 %. Na2CO3-O2 pretreatment is considered to be an operable and cost-efficient biorefining process and represents a promising method for the value-added utilization of WWS.

Improved Sugar Recovery from Orange Peel by Statistical Optimization of Thermo-Alkaline Pretreatment

Orange peel, which is a by-product of oranges, contains carbohydrates that can be converted into sugars and used in the fermentation process. In this study, the thermal alkaline pretreatment process was chosen because of its simplicity and lesser reaction time. In addition, the reaction factors were optimized using response surface methodology. The determined optimal conditions were as follows: 60.1 g/L orange peels loading, 3% KOH and 30 min. Under the optimal conditions, glucan content (GC) and enzymatic digestibility (ED) were found to be 32.8% and 87.8%, respectively. Enzymatic hydrolysis was performed with pretreated and non-pretreated orange peels using three types of enzyme complex (cellulase, cellobiase and xylanase). The minimum concentrations of enzyme complex required to obtain maximum ED were 30 FPU (filter paper unit), 15 CBU (cellobiase unit), and 30 XNU (xylanase unit) based on 1 g-biomass. Additionally, ED of the treated group was approximately 3.7-fold higher than that of the control group. In conclusion, the use of orange peel as a feedstock for biorefinery can be a strategic solution to reduce wastage of resources and produce sustainable bioproducts.

Alkaline incubation improves the saccharification of poplar after sodium chlorite pretreatment with ultra-low cellulase loading

The limitation of xylan hydrolysis is a characteristic problem of sodium chlorite (SC) pretreatment, which greatly restricted the saccharification of SC-pretreated lignocelluloses. In this work, the effect of alkali incubation on the xylan hydrolysis of SC-pretreated poplar was investigated. SC pretreatment of poplar removed 84.7% lignin, which yielded 71.2% glucose and 60.0% xylose under a cellulase loading of 20 filter paper units (FPU)/g dry matter (DM). The subsequent alkali incubation reduced the acetyl group, surface lignin, and surface chlorine contents of SC-pretreated poplar. Glucose and xylose yields were increased to 92.4% and 86.3%, respectively, by incubating SC-pretreated samples with 1.0% NaOH at 50 °C for 1 h using a cellulase loading of 2 FPU/g DM. Both Tween 80 and xylanase improved the enzymatic hydrolysis yield of SC-pretreated poplar. Alkali incubation showed a 9-fold decrease in cellulase loading and increased the xylose yield of SC-pretreated poplar by 119.8%. This work overcame the limitation of xylan hydrolysis in sodium chlorite-pretreated lignocelluloses and represented a novel method to efficiently saccharification of poplar with ultra-low cellulase loading.

High-efficient cellulosic butanol production from deep eutectic solvent pretreated corn stover without detoxification

Biobutanol is one of the most promising renewable biofuels, which can be produced from lignocellulosic biomass by Clostridium. However, due to the dense structure of lignin, biomass is difficult to be hydrolyzed into fermentable sugars. This study has developed a short time (90 min) and low energy consumption (120 °C) pretreatment process using deep eutectic solvent (DES). Importantly, the cellulose recovery of treated corn stover (CS) after DES pretreatment at 120 °C was 99.24 %, while the removal of xylan and lignin reached 57.58 % and 61.40 %, respectively. The total sugars released from treated CS at 120 °C reached 37.94 g L−1 (glucose: 30.59 g L−1; xylose: 7.35 g L−1) at a low enzyme loading of 7.5 filter paper unit (FPU) g−1 pretreated CS. Furthermore, the DES treated CS hydrolysate was successfully used to produce butanol by Clostridium beijerinckii P260 without any detoxification and sterilization, thus producing 13.65 g L-1 total acetone-butanol-ethanol (ABE) with a high productivity (0.68 g L-1 h−1) and yield (0.38 g g−1). Overall, the present work provides a new insight into the efficient application of DES systems in lignocellulose biomass biorefinery.

Cellulose solvent–based pretreatment and enzymatic hydrolysis of pineapple leaf waste biomass for efficient release of glucose towards biofuel production

Leaf waste biomass of pineapple Ananas comosus can be used as the substrate for the second-generation biofuel production. The non-environmental friendly practice of pineapple leaf waste handling on the field can largely be avoided by directing the leaf waste as a feedstock for biofuels. In this study, three varieties of pineapple queen, simanchal (a local variety of India), and kew were used for the production of glucose. The leaf biomass of these pineapple varieties contained 19 to 22% extractives, 31 to 32% glucan, 17 to 22% hemicellulose, and 17 to 20% lignin. Biomass without any pre-treatment when digested with cellulase released 36 to 43% glucose, whereas glucose release percentages significantly elevated when the biomass was subjected to phosphoric acid-based cellulose solvent and organic solvent lignocellulose fractionation (COSLIF) pretreatment before hydrolysis. Herein, we found that cellulase enzyme dosages 5 filter paper unit (FPU) along with 10 international units (IU) of β-glucosidase per gram of glucan of COSLIF-pretreated leaf biomass of queen variety resulted in the generation of 84% glucose after 72 h of incubation. Assuming 100% conversion of glucose to ethanol, theoretical ethanol yield was calculated to be 212 L from 1 t dry leaf biomass.

Optimizing the saccharification of pretreated wood biomass using crude enzyme from Acanthophysium sp. KMF001

This study was conducted on crude enzyme from the novel strain Acanthophysium sp. KMF001 using a mediator (surfactant). The surfactant was applied to the steam-exploded pretreated domestic wood biomass, and response surface methodology (RSM) was conducted to determine the optimum conditions for saccharification using the optimum substrate concentration, enzyme concentration, and surfactant concentration. Steam-explosion of Korean oak (25 kgf/cm2) for 7 min showed a maximum-predicted saccharification of approximately 99.9% at 7.0% substrate concentration, 37.5 FPU (filter paper units) enzyme concentration, and 475.8 mg/g-glucan surfactant (polysorbate 80) concentration. Steam-explosion of red pine (25 kgf/cm2) for 7 min revealed a maximum prediction saccharification rate of approximately 58.7% at 6.5% substrate concentration, 36.3 FPU enzyme concentration, and 330.3 mg/g-glucan surfactant (polysorbate 80) concentration. The extents of saccharification of Korean oak (99.9%) and pine (58.7%) demonstrated the high applicability of the crude enzyme from Acanthophysium sp. KMF001.