Cellulase Preparation Research Articles

Study on screening of fermentation agents and optimization of the fermentation process for pharyngitis tablet residue.

This study aimed to screen an appropriate starter and determine the optimal fermentation process parameters to optimize the fermentation process of nutrient components and bioactive contents in pharyngitis tablet residue. This study included two experiments. In experiment I, single-factor experimental design was used to study the effects of different biological starters (cellulase preparation; Lactobacillus Plantarum and Bacillus subtilis preparation; mixture of cellulase + Lactobacillus Plantarum and Bacillus subtilis) on the nutritional values and bioactive ingredient contents in pharyngitis tablet residue. In experiment II, orthogonal design experiment was adopted to study the effects of initial water content (45, 50, and 55%), fermentation temperature (35, 37, and 40°C), and fermentation time (24, 48, and 72 h) on the changes in nutrient components, biologically activity contents, and toxin contents of the residue after optimal fermentation agent treatment. Cellulase preparation was found to be the optimal starter. The optimal fermentation conditions were: initial water content, 55%; fermentation temperature, 37°C; and fermentation time, 72 h. The contents of aflatoxin B1 and vomit toxin were in line with Chinese feed hygiene production standards. The fermentation quality of pharyngitis tablet residue can be improved by using the optimal starter and fermentation conditions.

Optimized Cellulase-Hydrolyzed Deoiled Coconut Cake Powder as Wheat Flour Substitute in Cookies.

Deoiled coconut cake powder (DCCP) was hydrolyzed to reduce the ratio of insoluble/soluble dietary fiber (RIS) by partially converting insoluble dietary fiber to soluble using Celluclast 1.5 L, a commercial cellulase preparation in citrate buffer medium. Firstly, the influence of citrate buffer amount, enzyme concentration, pH, and retention time on the enzymatic hydrolysis efficiency was investigated. Then, response surface methodology (RSM) was employed to optimize the process in which the insoluble and soluble dietary fiber contents were the responses. The results revealed that 10.3 g buffer/g of materials, 3.7 U/g of the materials, and 60 min of retention time were the optimal conditions for the enzymatic hydrolysis to obtain the insoluble and soluble contents of 68.21%db and 8.18%db, respectively. Finally, DCCP or hydrolyzed DCCP (HDCCP) was partially substituted for wheat flour at different replacement ratios in a cookie recipe at 0, 10, 20, 30, and 40%. The cookies with a 10% replacement ratio of hydrolyzed deoiled coconut cake powders had a lower RIS by more than two folds those of DCCP and had the same sensorial score as the control sample. This study proposed that Celluclast 1.5 L effectively reduced RIS by partially converting insoluble to soluble dietary fiber, improving the soluble dietary fiber content in fiber-enriched cookies.

RETRACTED ARTICLE: Preparation of carrierized cellulase Cellulase@MIL-88B(Fe) and its enzymatic properties

We, the Editors and Publisher of Green Chemistry Letters and Reviews have retracted the following article: Erhong Zhang, Meng Wang, Xiaoliang Pan, Xinfeng Wang & Wenju Zhang (2022) Preparation of carrierized cellulase Cellulase@MIL-88B(Fe) and its enzymatic properties, Green Chemistry Letters and Reviews, 15:3, 627-637, DOI: https://doi.org/10.1080/17518253.2022.2124890 Following publication, the authors requested the withdrawal of this article in 2022 due to a conflict with an associated patent. Further investigations by the Publisher, in close collaboration with the Editor-in-Chief, could not confirm a conflict with an associated patent but revealed that the article has significant overlap with a chapter of an unpublished thesis titled “Construction of an adaptive immobilized cellulase system and its enzymatic hydrolysis performance” authored by Tiantian Li in June 2022. More specifically, the overlap involves most of the data in the article which was reused without the required permissions, or the knowledge of the owners of the data. Upon query, the authors have cooperated with the investigation, but could neither demonstrate that their data was obtained appropriately, nor that their study was conducted in compliance with our editorial policies. As the reuse of data from a third-party without acquiring the required permissions is a serious breach of publishing ethics, we are retracting the article from the journal. The corresponding author listed in this publication has been informed. We have been informed in our decision-making by our editorial policies and the COPE guidelines. The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as ‘Retracted’.

Amperometric method for the determination of cellulase activity and its optimization using response surface method

Cellulases are a group of enzymes, which catalyse different steps of cellulose hydrolysis, and are broadly used in industry as unpurified mixtures of several enzymes. The total activity of cellulase is defined as the ability of the enzyme to produce glucose, which is the final product of cellulose hydrolysis, and is expressed in cellulase units. However, common strategies for the determination of the cellulolytic activity of industrial cellulase preparations are based on the assessment of different steps of cellulose hydrolysis, and the results obtained with different methods are not similar. The aim of the present study was to develop an assay for the determination of cellulase activity that relies on the amperometric determination of the final product of cellulose hydrolysis glucose. The assay conditions were optimized using response surface methodology (RSM) combined with Box-Behnken design. The detection limit of the proposed method was 1.71 ± 0.06 U. We compared the results of the amperometric method with the ones obtained with the spectrophotometric method and viscosimetry in a commercial cellulose preparation.

Enhancing cellulases through synergistic β-glucosidases for intensifying cellulose hydrolysis

β-glucosidases (BGs) are key enzymes in cellulose hydrolysis, being the rate-limiting enzyme that is regulated by feedback inhibition from its own product—glucose. Due to which low hydrolysis yields are observed thus affecting overall ethanol yield per ton biomass. In this study, hydrolysis yields influence as a function of BG deficiency, supplementation, and its synergy in cellulase cocktails were investigated. Results show that commercial cellulase preparations were low in BG and produce hydrolysis yields below 60 % in 12 h. On blending commercial preparations with BGs (from fungal and bacterial sources), the degree of synergism was observed greater than 1, indicating that the components depicted synergistic behaviour in the mixture. A triple blend cellulase cocktail comprising commercial preparation, and two different BG preparations were optimized using the mixture design approach based on the synergism demonstrated. According to the findings, the in-house optimized cocktail (C-Mix) was able to hydrolyze six different two-stage treated LBM feedstocks and release more than 900 mg/g of glucose. The stability of the optimized cocktail was also enhanced two-fold due to strong synergistic interactions. Thus, a simple blending of BGs in the commercial preparations can be a sustainable approach for developing effective cocktails and intensifying cellulose hydrolysis.

Justification of an approach to cellulase application in enzymatic softening of linen fabrics and clothing

We propose the final softening finishing of textile products based on a complex of modifying effects on a highly ordered cellulose backbone of the secondary cell wall G-layer or on a mesh corset of microfibrillar cellulose in the primary cell wall. We suggest using polypeptide molecule sizes and the strength of polypeptide molecule adsorption binding on a model solid-phase substrate as parameters of the structurally regulated action of cellulase preparations. The article presents an analysis of 11 cellulase preparations as groups of weakly, tightly and medium adsorption enzymes, and also groups of volumetric, peripheral or combined activity manifested in the fiber structure. We show the regularities of enzyme property influence on the fiber pore structure development, changes in the parameters of the fabric bending stiffness and breaking load and technological shrinkage for two modification modes: liquid treatment with flax fiber swelling or low-modulus application solution. For the liquid method of treatment (with a bath module of at least 10) it is recommended to apply highly adsorbable cellulases ( A > 50%), in which the content of molecules with a globule size less than 30 nm does not exceed 15%. For the low-modulus method of treatment it is recommended to use enzymes with the adsorption capacity of at least 40% and a low content of small-sized fractions (no more than 25%). The recommended treatment variants ensure a three to four-fold bending stiffness decrease at the mechanical strength loss of not more than 15% and shrinkage of linen fabrics not more than 0.5%.

Aspergillus-Derived Cellulase Preparation Exhibits Prebiotic-like Effects on Gut Microbiota in Rats

Aspergillus-derived cellulase, which is involved in the production of Aspergillus-fermented foods, has been employed in the food and animal feed industries. However, the effect of dietary Aspergillus cellulase on health is poorly understood. Previously, we discovered that supplemental Aspergillus-derived protease and lipase preparations had substantial bifidogenic effects on rats fed a high-fat diet. Therefore, this study reports on the effects of a 0.1% dietary Aspergillus-derived cellulase preparation (CEL) on the gut microbiota of rats fed a high-fat diet. Gene sequencing analysis of 16S rRNA revealed that CEL treatment markedly affected the microbiota profiles of the cecal contents (p < 0.05). Notably, CEL markedly increased the relative abundance (RA) of typical probiotics, such as Bifidobacterium and Lactobacillus, at the genus level (28- and 5-fold, respectively, p < 0.05). Similarly, at the family level, CEL treatment significantly increased the RA of Bifidobacteriaceae and Lactobacillaceae (p < 0.05). Furthermore, CEL increased the RA of other genera, such as Collinsella and Enterococcus, but decreased the RA of Oscillospira, Dorea and Coprobacillus (p < 0.05). The effects on these genera are similar to those reported for typical prebiotic oligosaccharides. Overall, this study demonstrates the prebiotic-like effects of dietary CEL by significantly increasing Bifidobacterium and Lactobacillus abundance.

Preparation and identification of carboxymethyl cellulose-degrading enzyme candidates for oilfield applications

Carboxymethyl cellulose (CMC) is often used during hydraulic fracturing (fracking) operations as a fluid viscosifier to facilitate proppant delivery. However, the accumulation of residual CMC at fracture faces can result in formation damage, thereby impeding oil and gas recovery. Whereas harsh chemical oxidizers are typically added to disrupt these polymer accumulations, there is now industrial interest in developing clean, biological approaches for the degradation of CMC under fracking conditions. Using a methanogenic culture known to utilize CMC under conditions typically found in oil fields, we developed an efficient method to isolate and purify CMC-degrading enzymes. Initial purification and concentration of cellular components produced an increase in exo-ß-(1,4)-exoglucanase and ß-(1,4)-glucosidase activities by 9-fold and 26-fold, respectively. Partially purified extracts provided substantial degradation of CMC as monitored by viscosity reduction within three hours at 50 °C, an improvement over the untreated cell-free extract which required 48 h to achieve similar viscosity values, outperforming a commercially-available cellulase preparation. Putative cellulases were identified within the isolated enzyme population, with endo-ß-(1,4)-xylanase from Caldicoprobacter faecalis hypothesized to be an important contributor to CMC degradation. This study demonstrates that enzyme technology holds great promise as a viable approach to degrade CMC accumulations under field conditions.

Constitutive expression of codon optimized Trichoderma reesei TrCel5A in Pichia pastoris using GAP promoter

To address the deficient activity of TrCel5A in naturally secreted cellulase preparation, this study used the GAP promoter to induce constitutive expression of Trichoderma reesei TrCel5A in Pichia pastoris. A recombinant TrCel5A was screened out after gene optimization, synthesis, and expression. The biochemical and enzymatic properties of the new recombinant were characterized. As a result, optimization of shake-flask fermentation of the recombinant was obtained at 28 °C, 2% inoculum volume, an initial pH of 6.0, as well as glycerol and Tween-80 additions of 30 g/L and 6 g/L, respectively. Under the above-optimized conditions, the recombinant produced 14.8 U/mL of the enzyme activity at 96 h of fermentation. To further enhance enzyme production, pilot-scale cultivation was evaluated using 5-L bioreactors. Using high-cell-density fermentation, the recombinant strain increased enzyme activity to 130.4 U/ml and protein content to 2.49 g/L. In addition, the kinetic factors, including Km and Vmax values for TrCel5A, were detected to be 5.1 mg/mL and 265.9 μmol/(min.mg), respectively. Thus, TrCel5A was effectively expressed in P.pastoris under the GAP promoter, and it demonstrated its potential in commercially relevant enzyme hydrolysis of lignocellulosic biomass.

Effects of cellulolytic treatment conditions on dietary fiber content of grape pomace and use of enzyme-treated pomace in cookie making

Grape pomace is a rich source of dietary fibers and phenolic compounds; it has been added to bakery products to enhance their dietary fiber content and antioxidant activity. Nevertheless, the Insoluble Dietary Fiber (IDF) content of grape pomace is significantly higher than the Soluble Dietary Fiber (SDF) content, the IDF/SDF ratio of bakery products was much higher than the recommended value of 3:1 from the dietetic associations. In this research, grape pomace was treated with cellulase preparation to partially convert IDF to SDF. The appropriate conditions of the cellulolytic treatment were as follows: initial moisture content of grape pomace of 7.5 g water/g dry basis, cellulase concentration of 6 U/g dry basis and treatment time of 1 h. Under these conditions, the SDF content increased by 20% and the IDF/SDF ratio decreased by 23.2%. Then, the enzyme-treated grape pomace (ETGP) and untreated grape pomace (UGPM) were separately mixed with wheat flour for cookie formulation; the grape pomace ratio was 20% of the composite flour weight. The use of ETGP or UTGP significantly improved the content of total, insoluble and soluble dietary fiber, total phenolics, anthocyanin of cookies as well as their antioxidant activity measured by DPPH and FRAP assays. Cookies supplemented with ETGP or UTGP had increased hardness and reduced brightness. The enzymatic treatment of grape pomace did not affect the total dietary fiber and antioxidant activity of cookies; however, their IDF/SDF ratio decreased to 12% and the value of 3.7 was close to the recommended ratio from the dietetic associations. Furthermore, the preference scores of ETGP added cookies and UTGP added cookies were higher than that of the control cookies without grape pomace addition.

Optimization of juice production from “cempedak” (Artocarpus integer) fruit pulp liquefied with the aid of enzymes

PurposeThe purpose of this research is to produce “cempedak” juice using enzyme aided-liquefaction by examining the effects of enzyme types (Pectinex® Ultra SP-L, Celluclast® 1.5 L and Fungamyl® 800 L), enzyme concentrations (0–1.5% v/w), incubation time (0–2.5 h) and incubation temperature (35–60 °C) on juice yield and viscosity, total soluble solids and color of fruit puree.Design/methodology/approachRipe “cempedak” pulp from CH28 fruit was first pureed in a blender and then homogenized with water at 1:2 ratio. The diluted puree was then liquefied with the enzymes separately to reduce its viscosity. Analyses such as juice yield, viscosity, total soluble solids and color of the liquefied “cempedak” puree were then carried out.FindingsResults indicated that the optimized use of 1.2% (v/w) Celluclast® 1.5 L (Novozymes, Denmark), a cellulase preparation, at 45 °C and 1 h produced juice with the lowest viscosity (349.4 cP) and the highest juice yield (82.3% v/w). Liquefied “cempedak” juice was darker (with L* value of 51.17) and more yellowish (b* value of 38.88) compared to “cempedak” juice without liquefaction (control). When compared to untreated “cempedak” juice, the droplet size of “cempedak” juice obtained after liquefaction under optimized conditions was found to be lower, regardless of whether the juice was filtered (with a total reduction of 23% of droplet size) or not filtered (with a total reduction of 16% of droplet size). The results indicate the possibility of employing Celluclast® 1.5 L to produce “cempedak” juice that can be further processed such as for the production of “cempedak” fruit powder.Originality/valueThis paper provides information on the enzyme concentration, incubation time and temperature for liquefying “cempedak” pulp such that the liquefied material produced can be used as a base feed for spray-drying to produce “cempedak” fruit powder.

Expression and production of thermophilic alginate lyases in Bacillus and direct application of culture supernatant for seaweed saccharification

Brown seaweeds are rich in carbohydrates and may be used as a source of fermentable sugars. Saccharification of the seaweed biomass can be carried out enzymatically by a combination of cellulases and alginate lyases. In this study, thermotolerant exo- and endo alginate lyases were cloned and expressed in Bacillus subtilis. The lyases were secreted to the culture supernatant and used directly together with a commercial cellulase preparation to saccharify Saccharina latissima biomass. The results showed that the strategy of using the culture supernatants directly as a source of alginate lyases worked very well, releasing glucose, mannitol, and uronic acids. The ratio between the exo- and endo-acting alginate lyases proved to be very important for saccharification yield, and under optimal reaction conditions the use of culture supernatants containing alginate lyases improved final glucose concentration by 73%, when compared to only applying cellulases. This direct use of culture supernatants as a source of alginate lyases shows that enzyme purification steps are not needed, saving seaweed processing costs and points to the possibility of a relatively simple on-site enzyme production for seaweed biorefining.

Efficiency enhancement of a new cellulase cocktail at low enzyme loading for high solid digestion of alkali catalyzed atmospheric glycerol organosolvent pre-treated sugarcane bagasse

The acquisition during biomass saccharification of elevated levels of fermentable sugars with lower cellulase concentration is central to ensuring an economically viable and industrially relevant hydrolytic process. Thus, using a new cellulase preparation (LT4) at low cellulase loading (2 mg protein/g dried substrate), this study assessed the possible boosting effect of integrating accessory enzymes and additives on high-solids hydrolysis of sugarcane bagasse via fed-batch feeding. Hydrolysis which commenced with initial 8% solids loading and subsequent substrate feeding of 4% solids at 6 h, 18 h, and 24 h respectively, proved optimal for the 20% high-solids saccharification producing 158 g/L total sugars and 83% glucose yield after 72 h with the combined optimized additives and accessory enzymes. The results obtained indicate that the integration of accessory enzymes and additives offers a benignant approach to minimizing the enzyme load and cost of high solids saccharification of lignocellulosic heteropolymers while also boosting enzyme hydrolytic performance.

Improvements of the productivity and saccharification efficiency of the cellulolytic \u03b2-glucosidase D2-BGL in Pichia pastoris via directed evolution

Backgroundβ-Glucosidases are essential for cellulose hydrolysis by catalyzing the final cellulolytic degradation of cello-oligomers and cellobiose to glucose. D2-BGL is a fungal glycoside hydrolase family 3 (GH3) β-glucosidase isolated from Chaetomella raphigera with high substrate affinity, and is an efficient β-glucosidase supplement to Trichoderma reesei cellulase mixtures for the saccharification of lignocellulosic biomass.ResultsWe have carried out error-prone PCR to further increase catalytic efficiency of wild-type (WT) D2-BGL. Three mutants, each with substitution of two amino acids on D2-BGL, exhibited increased activity in a preliminary mutant screening in Saccharomyces cerevisiae. Effects of single amino acid replacements on catalysis efficiency and enzyme production have been investigated by subsequent expression in Pichia pastoris. Substitution F256M resulted in enhancing the tolerance to substrate inhibition and specific activity, and substitution D224G resulted in increasing the production of recombinant enzyme. The best D2-BGL mutant generated, Mut M, was constructed by combining beneficial mutations D224G, F256M and Y260D. Expression of Mut M in Pichia pastoris resulted in 2.7-fold higher production of recombinant protein, higher Vmax and greater substrate inhibition tolerance towards cellobiose relative to wild-type enzyme. Surprisingly, Mut M overexpression induced the ER unfolded protein response to a level lower than that with WT D2 overexpression in P. pastoris. When combined with the T. reesei cellulase preparation Celluclast 1.5L, Mut M hydrolyzed acid-pretreated sugarcane bagasse more efficiently than WT D2.ConclusionsD2-BGL mutant Mut M was generated successfully by following directed evolution approach. Mut M carries three mutations that are not reported in other directed evolution studies of GH3 β-glucosidases, and this mutant exhibited greater tolerance to substrate inhibition and higher Vmax than wild-type enzyme. Besides the enhanced specific activity, Mut M also exhibited a higher protein titer than WT D2 when it was overexpressed in P. pastoris. Our study demonstrates that both catalytic efficiency and productivity of a cellulolytic enzyme can be enhanced via protein engineering.

Progress in the production of lignocellulolytic enzyme systems using Penicillium species

The efficient production of lignocellulolytic enzyme systems is an important support for large-scale biorefinery of plant biomass. On-site production of lignocellulolytic enzymes could increase the economic benefits of the process by lowering the cost of enzyme usage. Penicillium species are commonly found lignocellulose-degrading fungi in nature, and have been used for industrial production of cellulase preparations due to their abilities to secrete complete and well-balanced lignocellulolytic enzyme systems. Here, we introduce the reported Penicillium species for cellulase production, summarize the characteristics of their enzymes, and describe the strategies of strain engineering for improving the production and performance of lignocellulolytic enzymes. We also review the progress in fermentation process optimization regarding the on-site production of lignocellulolytic enzymes using Penicillium species, and suggest prospect of future work from the perspective of building a "sugar platform" for the biorefinery of lignocellulosic biomass.

Effects of enzymatic treatment conditions on dietary fibre content of wheat bran and use of cellulase‐treated bran in cookie

SummaryA commercial cellulase preparation (Viscozyme Cassava C) was employed to process wheat bran to improve the ratio of insoluble to soluble dietary fibre. The effects of initial moisture content, enzyme dosage, and incubation time on the content of soluble and insoluble fibre during the enzymatic treatment were investigated. The appropriate conditions for the cellulase treatment were initial moisture content of 0.75 g water g−1 dry matter, enzyme dosage of 9 U g−1 dry matter and incubation time of 120 min under which the ratio of insoluble to soluble fibre of wheat bran was lowered by 42%. Untreated and cellulase‐treated wheat bran was incorporated into cookies at levels of 0%, 20%, 30%, 40% and 50%. Cookie with 50% cellulase‐treated bran showed 21% lower ratio of insoluble to soluble fibre as well as 14% lower hardness and 13% higher overall acceptability than sample with the same amount of untreated bran.

Efficient Constitutive Expression of Cellulolytic Enzymes in Penicillium oxalicum for Improved Efficiency of Lignocellulose Degradation.

Efficient cellulolytic enzyme production is important for the development of lignocellulose-degrading enzyme mixtures. However, purification of cellulases from their native hosts is time- and labor-consuming. In this study, a constitutive expression system was developed in Penicillium oxalicum for the secreted production of proteins. Using a constitutive polyubiquitin gene promoter and cultivating with glucose as the sole carbon source, nine cellulolytic enzymes of different origins with relatively high purity were produced within 48 h. When supplemented to a commercial cellulase preparation, cellobiohydrolase I from P. funiculosum and cellobiohydrolase II from Talaromyces verruculosus showed remarkable enhancing effects on the hydrolysis of steam-exploded corn stover. Additionally, a synergistic effect was observed for these two cellobiohydrolases during the hydrolysis. Taken together, the constitutive expression system provides a convenient tool for the production of cellulolytic enzymes, which is expected to be useful in the development of highly efficient lignocellulose-degrading enzyme mixtures.

The effect of fermentation time and addition of crude cellulase to concentration of bioethanol in bagasse fermentation

Bagasse is a solid waste from the sugar cane milling process in the sugar industry. Bagasse contains lignin, cellulose and hemicellulose, which through the fermentation process by Phanerochaete chrysosporium can produce crude cellulase, furthermore cellulase and other enzymes can be used in bagasse fermentation to produce bioethanol. The purpose of this study was to determine the effect of fermentation time and the addition of crude cellulase to the yield and concentration of bioethanol produced in bagasse fermentation. The research was carried out with the stages of the process: Preparation of raw materials (bagasse), Preparation of crude cellulase, Fermentation process, Product analysis (Bioethanol). The variables used in the experiment were fermentation time (96, 120, and 144 hours) and the addition of crude cellulase (10%, 20%, 30%, 40%, and 50% (v/v)). Analysis of crude cellulase activity using the DNS method, while the analysis of bioethanol concentration using the chromatography methods. The results showed that the maximum yield was 16.24% and the highest bioethanol concentration of 11.04% was obtained at the time of fermentation of 144 hours and the addition of crude cellulase by 50% (v/v).

Pasta from cellulase-treated wheat bran and durum semolina: Effects of vital gluten addition and/or transglutaminase treatment

Wheat bran was treated with a cellulase preparation and used in the production of high fiber pasta by the partial substitution of durum semolina. The nutritional, textural, and cooking quality of bran-incorporated pasta was measured. In addition, the potentials of vital gluten addition and/or transglutaminase treatment in improving the quality of bran-incorporated pasta were investigated. The use of cellulase-treated wheat bran resulted in a higher soluble fiber content, a more balanced insoluble to soluble fiber ratio, and a lower predicted glycemic index of pasta compared to that of untreated bran. The addition of vital gluten or treatment with transglutaminase improved the cooking performance, textural properties, and overall acceptability of pasta. In particular, the combination of vital gluten addition and transglutaminase treatment was more effective in improving the quality of pasta with cellulase-treated wheat bran.

Improving the fermentable sugar yields of wheat straw by high-temperature pre-hydrolysis with thermophilic enzymes of Malbranchea cinnamomea

BackgroundEnzymatic hydrolysis is a key step in the conversion of lignocellulosic polysaccharides to fermentable sugars for the production of biofuels and high-value chemicals. However, current enzyme preparations from mesophilic fungi are deficient in their thermostability and biomass-hydrolyzing efficiency at high temperatures. Thermophilic fungi represent promising sources of thermostable and highly active enzymes for improving the biomass-to-sugar conversion process. Here we present a comprehensive study on the lignocellulosic biomass-degrading ability and enzyme system of thermophilic fungus Malbranchea cinnamomea N12 and the application of its enzymes in the synergistic hydrolysis of lignocellulosic biomass.ResultsMalbranchea cinnamomea N12 was capable of utilizing untreated wheat straw to produce high levels of xylanases and efficiently degrading lignocellulose under thermophilic conditions. Temporal analysis of the wheat straw-induced secretome revealed that M. cinnamomea N12 successively degraded the lignocellulosic polysaccharides through sequential secretion of enzymes targeting xylan and cellulose. Xylanase-enriched cocktail from M. cinnamomea N12 was more active on native and alkali‑pretreated wheat straw than the commercial xylanases from Trichoderma reesei over temperatures ranging from 40 to 75 °C. Integration of M. cinnamomea N12 enzymes with the commercial cellulase preparation increased the glucose and xylose yields of alkali‑pretreated wheat straw by 32 and 166%, respectively, with pronounced effects at elevated temperature.ConclusionsThis study demonstrated the remarkable xylanase-producing ability and strategy of sequential lignocellulose breakdown of M. cinnamomea N12. A new process for the hydrolysis of lignocellulosic biomass was proposed, comprising thermophilic enzymolysis by enzymes of M. cinnamomea N12 followed with mesophilic enzymolysis by commercial cellulases. Developing M. cinnamomea N12 as platforms for thermophilic enzyme mixture production will provide new perspectives for improved conversion yields for current biomass saccharification schemes.