Production Of Lignocellulolytic Enzymes Research Articles

Successive process for efficient biovalorization of Brewers’ spent grain to lignocellulolytic enzymes and lactic acid production through simultaneous saccharification and fermentation

This study aimed to increase the value of brewers’ spent grain (BSG) by using it as feedstock to produce lignocellulolytic enzymes and lactic acid (LA). Twenty-two fungal strains were screened for lignocellulolytic enzyme production from BSG. Among them, Trichoderma sp. showed the highest cellulase activity (35.84 ± 0.27 U/g-BSG) and considerably high activities of xylanase (599.61 ± 23.09 U/g-BSG) and β-glucosidase (16.97 ± 0.77 U/g-BSG) under successive solid-state and submerged fermentation. The processes were successfully scaled up in a bioreactor. The enzyme cocktail was recovered and characterized. The maximum cellulase and xylanase activities were found at pH 5.0 and 50 °C, and the activities were highly stable at pH 4–8 and 30–50 °C. The enzyme cocktail was applied in simultaneous saccharification and fermentation of acid-pretreated BSG for LA production. The maximum LA obtained was 59.3 ± 1.0 g/L. This study has shown the efficient biovalorization of BSG, and this approach may also be applicable to other agro-industrial wastes.

Effect of co-cultivation of white and brown rot species on basidiome production, lignocelluloytic enzyme activity and dye decolourisation

The aim of this work was studying the impact of co-cultivating two mushroom species: a white (Pleurotus albidus CLA 45) and a brown rot one (Laetiporus sulphureus BAFC 205) in substrates based on poplar or pine sawdust, on their lignocellulolytic enzyme production, yield values and basidiomes properties. Laetiporus sulphureus only developed basidiome primordia, but P. albidus monoculture and co-culture in pine sawdust achieved biological efficiencies of up to 50–55 %. Co-cultivation on diverse substrates rendered varied enzyme titers. Laccase and Manganese peroxidase titers were highest in pine co-culture and P. albidus poplar monoculture, respectively. Enzymatic extracts obtained from spent poplar substrate of dual cultures displayed potential for treating non-sterile textile-coloured effluents, achieving 35 % decolourisation after 120 h. The knowledge available on the effects of co-culture of white and brown rot fungi is still limited. This study represents an initial exploration of the interaction between them within intensive cultivation conditions.

A closed-loop strategy for on-site production of saccharolytic enzymes for lignocellulose biorefinery using internal lignocellulosic hydrolysates

The cost-effective production of lignocellulose-saccharolytic enzymes is crucial for the bioconversion of plant biomass into fuels and chemicals. The integrated mode for producing these enzymes using lignocellulosic feedstocks as natural inducers has a cost advantage; however, enzyme production level is limited because of the solid-state and complex composition of the raw materials. This study aimed to establish an innovative process for the integrated production of lignocellulolytic enzymes using lignocellulosic hydrolysate as the carbon source. The cellulase-producing workhorse Trichoderma reesei was reprogrammed by combinatorial engineering of transcription factors and the introduction of heterologous β-glucosidases. This enabled inducer-independent production of a complete lignocellulose-saccharolytic enzyme cocktail using lignocellulose-derived monosaccharides as sole carbon source. Via continuous feeding of the enzymatic hydrolysate of pretreated corn stover, crude enzymes with filter paper enzyme activity of 60.4 U/ml and β-glucosidase activity of 503 U/ml were produced. Crude fermentation broth can be used directly to prepare hydrolysates, enabling nex-round enzyme production by recycling 3.1% of the hydrolysate. These results demonstrate a closed-loop strategy for integrated enzyme production using internally produced lignocellulosic sugars from biorefinery plants.

Isolation, screening wood rot fungi from the tropical forest of Thailand and their lignocellulolytic enzyme production under solid-state fermentation using agricultural waste as substrate

Isolation, screening wood rot fungi from the tropical forest of Thailand and their lignocellulolytic enzyme production under solid-state fermentation using agricultural waste as substrate

Structure-guided engineering of transcriptional activator XYR1 for inducer-free production of lignocellulolytic enzymes in Trichoderma reesei

The filamentous fungus Trichoderma reesei is widely used for the production of lignocellulolytic enzymes in industry. XYR1 is the major transcriptional activator of cellulases and hemicellulases in T. reesei. However, rational engineering of XYR1 for improved lignocellulolytic enzymes production has been limited by the lack of structure information. Here, alanine 873 was identified as a new potential target for the engineering of XYR1 based on its structure predicted by AlphaFold2. The mutation of this residue to tyrosine enabled significantly enhanced production of xylanolytic enzymes in the medium with cellulose as the carbon source. Moreover, xylanase and cellulase production increased by 56.7- and 3.3-fold, respectively, when glucose was used as the sole carbon source. Under both conditions, the improvements of lignocellulolytic enzyme production were higher than those in the previously reported V821F mutant. With the enriched hemicellulases and cellulases, the crude enzymes secreted by the A873Y mutant strain produced 51 % more glucose and 52 % more xylose from pretreated corn stover than those of the parent strain. The results provide a novel strategy for engineering the lignocellulolytic enzyme-producing capacity of T. reesei, and would be helpful for understanding the molecular mechanisms of XYR1 regulation.

CRISPR/Cas9 mediated gene editing of transcription factor ACE1 for enhanced cellulase production in thermophilic fungus Rasamsonia emersonii

BackgroundThe filamentous fungus Rasamsonia emersonii has immense potential to produce biorefinery relevant thermostable cellulase and hemicellulase enzymes using lignocellulosic biomass. Previously in our lab, a hyper-cellulase producing strain of R. emersonii was developed through classical breeding and system biology approaches. ACE1, a pivotal transcription factor in fungi, plays a crucial role in negatively regulating the expression of cellulase genes. In order to identify the role of ACE1 in cellulase production and to further improve the lignocellulolytic enzyme production in R. emersonii, CRISPR/Cas9 mediated disruption of ACE1 gene was employed.ResultsA gene-edited ∆ACE1 strain (GN11) was created, that showed 21.97, 20.70 and 24.63, 9.42, 18.12%, improved endoglucanase, cellobiohydrolase (CBHI), β-glucosidase, FPase, and xylanase, activities, respectively, as compared to parental strain M36. The transcriptional profiling showed that the expression of global regulator (XlnR) and different CAZymes genes including endoglucanases, cellobiohydrolase, β-xylosidase, xylanase, β-glucosidase and lytic polysaccharide mono-oxygenases (LPMOs) were significantly enhanced, suggesting critical roles of ACE1 in negatively regulating the expression of various key genes associated with cellulase production in R. emersonii. Whereas, the disruption of ACE1 significantly down-regulated the expression of CreA repressor gene as also evidenced by 2-deoxyglucose (2-DG) resistance phenotype exhibited by edited strain GN11 as well as appreciably higher constitutive production of cellulases in the presence of glucose and mixture of glucose and disaccharide (MGDs) both in batch and flask fed batch mode of culturing. Furthermore, ∆ACE1 strains were evaluated for the hydrolysis of biorefinery relevant steam/acid pretreated unwashed rice straw slurry (Praj Industries Ltd; 15% substrate loading rate) and were found to be significantly superior when compared to the benchmark enzymes produced by parent strain M36 and Cellic Ctec3.ConclusionsCurrent work uncovers the crucial role of ACE1 in regulating the expression of the various cellulase genes and carbon catabolite repression mechanism in R. emersonii. This study represents the first successful report of utilizing CRISPR/Cas9 genome editing technology to disrupt the ACE1 gene in the thermophlic fungus R. emersonii. The improved methodologies presented in this work might be applied to other commercially important fungal strains for which genetic manipulation tools are limited.

Lignocellulose Degrading Weizmannia coagulans Capable of Enantiomeric L-Lactic Acid Production via Consolidated Bioprocessing

Second-generation lactic acid production requires the development of sustainable and economically feasible processes and renewable lignocellulose biomass as a starting raw material. Weizmannia coagulans MA42 was isolated from a soil sample in Chiang Mai province, Thailand and showed the highest production of L-lactic acid and lignocellulolytic enzymes (cellulase, β-mannanase, xylanase, β-glucosidase, β-mannosidase, and β-xylosidase) compared to other isolates. Weizmannia coagulans MA42 was able to grow, secrete lignocellulolytic enzymes, and directly produce L-lactic acid in the medium containing various lignocellulosic feedstocks as the sole carbon source. Moreover, L-lactic acid production efficiency was improved after the substrates were pretreated with diluted sulfuric acid and diluted sodium hydroxide. The highest L-lactic acid production efficiency of 553.4 ± 2.9, 325.4 ± 4.1, 326.6 ± 4.4, 528.0 ± 7.2, and 547.0 ± 2.2 mg/g total available carbohydrate was obtained from respective pretreated substrates including sugarcane bagasse, sugarcane trash, corn stover, rice straw, and water hyacinth. It is suggested that structural complexity of the lignocellulosic materials and properties of lignocellulolytic enzymes are the key factors of consolidated bioprocessing (CBP) of lignocellulosic feedstocks to lactic acid. In addition, the results of this study indicated that W. coagulans MA42 is a potent bacterial candidate for CBP of a variety of lignocellulosic feedstocks to L-lactic acid production; however, further bioprocess development and genetic engineering technique would provide higher lactic acid production efficiency, and this would lead to sustainable lactic acid production from lignocellulosic feedstocks.

Single Mutation in Transcriptional Activator Xyr1 Enhances Cellulase and Xylanase Production in Trichoderma reesei on Glucose.

To achieve cost-effective production of lignocellulolytic enzymes for biorefinery processes, engineering transcription factors represents a powerful strategy to boost cellulase and xylanase in Trichoderma reesei. In this study, a novel mutation (R434L) in xylanase regulator 1 (Xyr1) was identified based on the yeast one-hybrid screening system. The point mutation was located in the middle homology region of Xyr1 with unclear functions, indicating a significant role for this domain in tuning Xyr1 transactivation. When constitutively expressed in T. reesei Δxyr1 (OEXR434L), Xyr1R434L led to highly improved production of both cellulases and xylanases on glucose compared with a strain similarly expressing Xyr1 (OEX). The respective 0.8- and 0.7-fold increases in extracellular pNPCase and xylanolytic activity were further verified to result from the greatly elevated transcription of major cellulase and xylanase genes in OEXR434L. Moreover, the saccharification efficiency of corn stover with OEXR434L enzyme cocktails was enhanced by 21% compared with that of OEX.

Lignocellulolytic enzyme cocktail produced by plant endophytic Chaetomium globosum exhibits a capacity for high-efficient saccharification of raw rice straw

The aim of this work was to optimize the production of lignocellulolytic enzyme cocktail by a plant endophyte Chaetomium globosum DX-THS3 under solid-state fermentation, and to evaluate the saccharification efficiency of this enzyme cocktail on raw rice straw. The process variables, including substrate type, substrate size, initial moisture content of substrate, culture temperature and fermentative time, were optimized using One-factor at a time approach and Response surface methodology. The results showed that the maximum CMCase, FPase and xylanase production reached 237.04 U/g, 54.8 U/g, and 987.33 U/g, respectively. Meanwhile, the lignocellulolytic enzyme cocktail exhibited good thermal stability between 30 °C and 60 °C, and excellent tolerance to acidic pH values of 3–6. Subsequently, the saccharification of raw rice straw by the enzyme cocktail was performed. After 24 h hydrolysis of raw rice straw, high reducing sugars yield (294.36 mg/g) and hydrolytic rate (47.58 %) were achieved. Furthermore, scanning electron micrograph also indicated that the enzyme cocktail had high hydrolysis performance for raw rice straw. Overall, our result provides a novel class of lignocellulolytic enzyme cocktail with a capacity for high-efficient and eco-friendly saccharification of raw rice straw, and strongly suggests plant endophytic fungi as excellent candidates for the production of lignocellulolytic enzymes.

Efecto del tamaño de partícula, aireación y altura del sustrato sobre la producción de enzimas lignocelulolíticas producidas por Trametes polyzona HHM001 crecido sobre residuos de hojas de maíz

The objective of the work was to evaluate the production of lignocllulolytic enzymes produced by Trametes polyzona HHM001 during its growth on corn leaf residues. Two particle sizes (PS) (PS8 and PS12), two levels of aeration (1 vvm and 0 vvm), as well as the height of the substrate in the production of lignocellulolytic enzymes were tested. The enzymatic activities of Laccase (Lcc), Lignin peroxidase (LiP), and Manganese peroxidase (MnP) were favored under aerated conditions (1 vvm). The enzymatic activity of Lcc was the most favored, with 80 Activity Units (AU)/mL, compared to the culture without aeration, in which 40 AU/mL was obtained. The production of xylanases (Xyl) and cellulases (Cel) were not influenced by aeration under the tested conditions. The results indicated that the particle size has more effect on enzyme production than the presence or absence of air, with a particle size of 8 where the best levels of enzymatic activity were observed. It was observed that the height of the substrate in the fermentation column strongly affects the activity of ligninolytic enzymes and not of the hydrolytic; at 5 cm, the highest ligninolytic activity was detected, where aeration favors oxidative conditions.

Conversion of lignocellulosic biomass: Production of bioethanol and bioelectricity using wheat straw hydrolysate in electrochemical bioreactor

The present study evaluated efficiency of wheat straw (WS) hydrolysate obtained through fungal pre-treatment to produce ethanol and electricity in an electrochemical bioreactor. Three white rot fungi Phanerochaete chrysosporium, Phlebia floridensis and Phlebia brevispora were used to degrade WS for hydrolysate preparation, Lignocellulolytic enzyme production was also monitored during the pretreatment. Yeast Pichia fermentans was allowed to ferment all three hydrolysates up to 12 days. The yeast showed maximum electrochemical response as open circuit voltage (0.672 V), current density 542.42 mA m−2, and power density of 65.09 mW m−2 on 12th day in the hydrolysate prepared using Phlebia floridensis. Maximum ethanol production of 9.2% (w/v) was achieved on 7th day with a fermentation efficiency of about 62.1%. Further, the coulombic efficiency improved from 0.06 to 1.46% during 12 days of the experiment. Thus, the results indicated towards the possible conversion of lignocellulosic biomass into bioethanol along with bioelectricity generation.

Characterization of fungi isolated from cattle dung with potential lignocellulolytic activities

Agricultural wastes not only serve as sources of environmental pollutions, but also serve as magnificent source of plant nutrients and natural repositories of bio-tools especially fungi of industrial interest for lignocellulolytic enzymes production. However, residues management particularly in rice–wheat system is a tedious phenomenon due to narrow gap between harvesting and sowing of these two crops. Hence, microbial intervention is one of the key options that may use successfully for their management. Thus, cellulolytic fungi were isolated from the cattle dung collected from various locations for cellulose manufacturing. The cellulolytic property of fungal isolates was confirmed by plate screening assay based on yellow zone formation surrounding of fungal culture on Congo red agar medium plate owing to production of cellulase enzyme. The colony diameter, clear zone diameter and cellulolytic index showing in descending order of CRD3, CRD5, CRD6 and CRD10 isolates, but –glucosidase activity was greater in CRD3 and CRD5 while, the lowest was in CRD9. However, isolates CRD7, CRD8 and CRD10 exhibited greater biochemical and enzymatic activity than rest of the promising fungal isolates. Similarly, isolates CRD1 and CRD2 utilized most of carbon sources as carbon substrate, but CRD7, CRD8 and CRD10 could not utilized mannitol and cellulose as carbon substrate. Overall, isolate CRD10 showed as a promising isolate in respect of biochemical, carbon utilization pattern and enzymatic activity than rest of the promising isolates.

Microbial Lignocellulolytic Enzymes for the Effective Valorization of Lignocellulosic Biomass: A Review

The urgent demand for alternative energy sources has been sparked by the tremendous burden on fossil fuels and the resulting acute energy crisis and climate change issues. Lignocellulosic biomass is a copious renewable and alternative bioresource for the generation of energy fuels and biochemicals in biorefineries. Different pretreatment strategies have been established to overcome biomass recalcitrance and face technological challenges, such as high energy consumption and operational costs and environmental hazards, among many. Biological pretreatment using microbial enzymes is an environmentally benign and low-cost method that holds promising features in the effective pretreatment of lignocellulosic biomass. Due to their versatility and eco-friendliness, cellulases, hemicellulases, and ligninolytic enzymes have been recognized as “green biocatalysts” with a myriad of industrial applications. The current review provides a detailed description of different types of lignocellulolytic enzymes, their mode of action, and their prospective applications in the valorization of lignocellulosic biomass. Solid state fermentation holds great promise in the microbial production of lignocellulolytic enzymes owing to its energy efficient, environment friendly, and higher product yielding features utilizing the lignocellulosic feedstocks. The recent trends in the application of enzyme immobilization strategies for improved enzymatic catalysis have been discussed. The major bottlenecks in the bioprocessing of lignocellulosic biomass using microbial enzymes and future prospects have also been summarized.

Tailoring the expression of Xyr1 leads to efficient production of lignocellulolytic enzymes in Trichoderma reesei for improved saccharification of corncob residues

BackgroundThe filamentous fungus Trichoderma reesei is extensively used for the industrial-scale cellulase production. It has been well known that the transcription factor Xyr1 plays an important role in the regulatory network controlling cellulase gene expression. However, the role of Xyr1 in the regulation of cellulase expression has not been comprehensively elucidated, which hinders further improvement of lignocellulolytic enzyme production.ResultsHere, the expression dosage of xyr1 was tailored in T. reesei by differentially overexpressing the xyr1 gene under the control of three strong promoters (Pegl2, Pcbh1, and Pcdna1), and the transcript abundance of xyr1 was elevated 5.8-, 12.6-, and 47.2-fold, respectively. We found expression of cellulase genes was significantly increased in the Pegl2-driven xyr1 overexpression strain QE2X, whereas relatively low in the Pcbh1- and Pcdna1-driven overexpression strains. We also found that the Pegl2-driven overexpression of xyr1 caused a more significant opening of chromatin in the core promoter region of the prominent cellulase genes. Furthermore, the cellulase activity showed a 3.2-fold increase in the strain QE2X, while insignificant improvement in the Pcbh1- and Pcdna1-driven strains. Finally, the saccharification efficiency toward acid-pretreated corncob residues containing high-content lignin by the crude enzyme from QE2X was increased by 57.2% compared to that from the parental strain. Moreover, LC–MS/MS and RT-qPCR analysis revealed that expression of accessory proteins (Cip1, Cip2, Swo1, and LPMOs) was greatly improved in QE2X, which partly explained the promoting effect of the Pegl2-driven overexpression on enzymatic hydrolysis of lignocellulose biomass.ConclusionsOur results underpin that the precise tailoring expression of xyr1 is essential for highly efficient cellulase synthesis, which provide new insights into the role of Xyr1 in regulating cellulase expression in T. reesei. Moreover, these results also provides a prospective strategy for strain improvement to enhance the lignocellulolytic enzyme production for use in biorefinery applications.

Integrating 1G with 2G Bioethanol Production by Using Distillers’ Dried Grains with Solubles (DDGS) as the Feedstock for Lignocellulolytic Enzyme Production

First-generation (1G) bioethanol is one of the most used liquid biofuels in the transport industry. It is generated by using sugar- or starch-based feedstocks, while second-generation (2G) bioethanol is generated by using lignocellulosic feedstocks. Distillers’ dried grains with solubles (DDGS) is a byproduct of first-generation bioethanol production with a current annual production of 22.6 million tons in the USA. DDGS is rich in fiber and valuable nutrients contents, which can be used to produce lignocellulolytic enzymes such as cellulases and hemicellulases for 2G bioethanol production. However, DDGS needs a pretreatment method such as dilute acid, ammonia soaking, or steam hydrolysis to release monosaccharides and short-length oligosaccharides as fermentable sugars for use in microbial media. These fermentable sugars can then induce microbial growth and enzyme production compared to only glucose or xylose in the media. In addition, selection of one or more suitable microbial strains, which work best with the DDGS for enzyme production, is also needed. Media optimization and fermentation process optimization strategies can then be applied to find the optimum conditions for the production of cellulases and hemicellulases needed for 2G bioethanol production. Therefore, in this review, a summary of all such techniques is compiled with a special focus on recent findings obtained in previous pieces of research conducted by the authors and by others in the literature. Furthermore, a comparison of such techniques applied to other feedstocks and process improvement strategies is also provided. Overall, dilute acid pretreatment is proven to be better than other pretreatment methods, and fermentation optimization strategies can enhance enzyme production by considerable folds with a suitable feedstock such as DDGS. Future studies can be further enhanced by the technoeconomic viability of DDGS as the on-site enzyme feedstock for the manufacture of second-generation bioethanol (2G) in first-generation (1G) ethanol plants, thus bridging the two processes for the efficient production of bioethanol using corn or other starch-based lignocellulosic plants.

Genome sequence analysis of halophilic Luteibacter sp. CQ10 to prospect its dual roles in antioxidants production and lignocellulose degradation

Luteibacter is a bacterial genus affiliated to Rhodanobacteraceae family with five species reported up to date. Currently, limited information regarding this genus was revealed, especially on its genomic perspective and its potential application. Here, we present the whole genome sequence of Luteibacter sp. CQ10, which the strain was isolated from mangrove bark collected at Tanjung Piai National Park, Johor, Malaysia. The genome of Luteibacter sp. CQ10 contains 30 contigs, with a genome size of 4.87 Mbp, a GC content of 66.44 % and a total of 4197 predicted genes. The preliminary analysis elucidated that a total of 10 and 22 annotated genes were predicted to participate in biosynthesis of antioxidants and production of lignocellulolytic enzymes, respectively. These biomolecules are beneficial to the food and biorefinery industries.

Improvement of Lignocellulolytic Enzyme Production Mediated by Calcium Signaling in Bacillus subtilis Z2 under Graphene Oxide Stress.

An increase in exoenzyme production can be enhanced by environmental stresses such as graphene oxide (GO) stress, but the link between the two events is still unclear. In this work, the effect of GO as an environmental stress factor on exoenzyme (lignocellulolytic enzyme, amylase, peptidase, and protease) biosynthesis was investigated in Bacillus subtilis Z2, and a plausible mechanism by which cytosolic Ca2+ regulates lignocellulolytic enzyme production in B. subtilis Z2 subjected to GO stress was proposed. The filter paper-hydrolyzing (FPase [representing total cellulase]), carboxymethylcellulase (CMCase [representing endoglucanase]), and β-glucosidase activities and extracellular protein concentration of the wild-type strain under 10 μg/mL GO stress were 1.37-, 1.64-, 1.24-, and 1.16-fold those of the control (without GO stress), respectively. Correspondingly, the transcription levels of lignocellulolytic enzyme genes, cytosolic Ca2+ level, and biomass concentration of B. subtilis were all increased. With lignocellulolytic enzyme from B. subtilis used to hydrolyze alkali-pretreated rice straw, the released reducing sugar concentration reached 265.53 mg/g, and the removal rates of cellulose, hemicellulose, and lignin were 52.4%, 30.1%, and 7.5%, respectively. Furthermore, transcriptome data revealed that intracellular Ca2+ homeostasis played a key role in regulating the levels of gene transcription related to the synthesis of lignocellulolytic enzymes and exoenzymes. Finally, the use of Ca2+ inhibitors (LaCl3 and EDTA) and deletion of spcF (a calmodulin-like protein gene) further demonstrated that the overexpression of those genes was regulated via calcium signaling in B. subtilis subjected to GO stress. IMPORTANCE To effectively convert lignocellulose into fermentable sugars, high lignocellulolytic enzyme loading is needed. Graphene oxide (GO) has been shown to promote exoenzyme (lignocellulolytic enzyme, amylase, peptidase, and protease) production in some microorganisms; however, the regulatory mechanism of the biosynthesis of lignocellulolytic enzymes under GO stress remains unclear. In this work, the lignocellulolytic enzyme production of B. subtilis under GO stress was investigated, and the potential mechanism by which B. subtilis enhanced lignocellulolytic enzyme production through the calcium signaling pathway under GO stress was proposed. This work revealed the role of calcium signaling in the production of enzymes under external environmental stress and provided a direction to facilitate lignocellulolytic enzyme production by B. subtilis.

Biotechnological valorization of oilseed cakes: Substrate optimization by simplex centroid mixture design and scale‐up to tray bioreactor

AbstractSunflower (SFC), rapeseed (RSC), and soybean cakes (SBC) are examples of oilseed cakes (OC) used in animal feed. Bioprocessing of these OC by solid‐state fermentation (SSF) aims to boost OC applications in feed and other industries by reducing antinutritional factors and releasing enzymes and antioxidants. A simplex centroid design was performed to optimize the substrate composition of SFC, RSC and SBC mixtures that maximize lignocellulolytic enzymes production by SSF with Aspergillus niger and consequently improve the nutritional properties of OC. Enzyme production by SSF of OC mixtures exceeded the activity values obtained using a single oilseed cake. A mixture composed of 50% (w/w) RSC and SBC was found to be the optimum substrate. The scale‐up from flasks to tray‐type bioreactors demonstrated SSF reproducibility, leading to the production of 299 U g−1 cellulase, 1476 U g−1 xylanase, 191 U g−1 β‐glucosidase and 220 U g−1 protease. The obtained enzymatic extract also presented antioxidant activity (around 50 μmol Trolox equivalents/g). This study demonstrated that bioprocessing of OC mixtures through SSF is an effective approach to obtain value‐added products with applications as feed or feed additives and to increase the liberation of bioactive compounds with applications in the feed, food, cosmetic and pharmaceutical industries. © 2022 Society of Chemical Industry and John Wiley & Sons, Ltd.

Valorization of agricultural residues using Myceliophthora thermophila as a platform for production of lignocellulolytic enzymes for cellulose saccharification

The use of lignocellulosic agricultural residues is emerging as a low-cost, sustainable, and eco-friendly approach for the production of microbial enzymes and other chemicals. In this study, the production of enzymes with lignocellulolytic potential and the secretome profile of the thermophilic fungus Myceliophthora thermophila were evaluated in solid-state cultivation on agricultural residues such as rice straw, soy straw, and wheat bran. In addition, the saccharification potential of the extracts was evaluated. The analysis of enzymatic activities (peptidase, lipase, amylase, CMCase, FPase, β-glucosidase, xylanase, pectinase, and laccase) and the 66 proteins identified in the secretome reinforced the hydrolytic and oxidative properties of the enzymatic lignocellulolytic arsenal produced by M. thermophila. The extracellular extract grown on rice straw obtained after 72 h of solid-state bioprocess achieved 46% saccharification of wheat bran substrate. Furthermore, the overall performance of the wheat bran extract (24 h) indicates the relevance of the CMCase, FPase and xylanase activities in saccharification processes. These results highlight the potential of M. thermophila as a platform for the production of enzymes with biotechnological potential, especially for lignocellulose degradation and cellulose saccharification. Solid-state bioprocess allows exploring agro-industrial residues, as a matrix and source of nutrients, for the enzyme production and emphasizes the possibility of adding value to agricultural waste. Moreover, the use of M. thermophila in solid-state bioprocesses can still be widely explored and optimized as a platform to scale up the production of cheapest fungal enzymes.

Production of Lignocellulolytic Enzymes and Phenolic Compounds by Lentinus strigosus from the Amazon Using Solid-State Fermentation (SSF) of Guarana (Paullinia cupana) Residue.

The Amazon rainforest has a rich biodiversity, and studies of Basidiomycete fungi that have biomolecules of biotechnological interest are relevant. The use of lignocellulosic biomass in biotechnological processes proposes an alternative use, and also adds value to the material when employed in the bioconversion of agro-industrial waste. In this context, this study evaluate the production of lignocellulolytic enzymes (carboxymethylcellulases (CMCase), xylanase, pectinase, laccase) as well as phenolic compounds and proteases by solid-state fermentation (SSF) using the fungus Lentinus strigosus isolated from Amazon. The guarana (Paullinia cupana) residue was characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). SSF was carried out with 60% humidification of the residue, at 30 °C, for 10 days. The lignocellulosic biomass presented fragmented structures with irregular shapes and porosities, and was mainly constituted by cellulose (19.16%), hemicellulose (32.83%), and lignin (6.06%). During the SSF, significant values of CMCase (0.84 U/g) on the 8th day, xylanase (1.00 U/g) on the 7th day, pectinase (2.19 U/g) on the 6th day, laccase (176.23 U/mL) on the 5th day, phenolic compounds (10.27 μg/mL) on the 1st day, soluble proteins (0.08 mg/mL) on the 5th day, and protease (8.30 U/mL) on the 6th day were observed. In general, the agro-industrial residue used provided promising results as a viable alternative for use as a substrate in biotechnological processes.