Highest Cellulase Production Research Articles

Screening, Gene Cloning and Expression of Cellulase-Producing Strain Bacillus subtilis Xh-16.

Cellulase is a complex enzyme system composed of multiple hydrolytic enzymes. It can degrade cellulose into glucose and improve the utilization efficiency of cellulose resources. Cellulase produced by microorganisms is the main method used in industry, offering the advantages of convenience and being environmentally friendly. A strain Xh-16 with high cellulase production was screened from the rotten rice straw. It was identified as Bacillus subtilis by morphological identification, physiological and biochemical identification, and 16S rRNA gene sequence analysis. Strain Xh-16 was used to degrade rice straw. After a 48h cellulase treatment, the complete degradation and structural breakdown of the straw were observed. We cloned the endoglucanase Cel5L gene of Bacillus subtilis Xh-16 and induced expression of the cloned gene in Escherichia coli BL21 (DE3). The results showed that the coding length of Cel5L gene was 1500bp, and the molecular weight of the encoded protein was about 55kDa. The molecular formula is C2456H3811N671O761S10 and it has 7709 atoms and 499 amino acids. Cel5L differs significantly from some the glycoside hydrolase family 5 cellulases because it has only one carbohydrate binding module family 3 at the C-terminus of its catalytic domain. The cellulase gene Cel5L in Xh-16 can encode active cellulase and can be heterologously expressed in Escherichia coli, which makes Escherichia coli have cellulase function. This study serves as a foundation for further research on cellulase diversity in Bacillus subtilis and offers insights for enhancing cellulase production.

Transcriptomic and metabolic changes in Trichoderma reesei caused by mutation in xylanase regulator 1 (xyr1)

BackgroundTrichoderma reesei is known for its ability to produce large amounts of extracellular proteins and is one of the most important industrially used filamentous fungus. Xylanase regulator 1 (XYR1) is the master regulator responsible for the activation of cellulase and hemicellulase gene expression under inducing conditions. It has been reported that strains with point mutations in certain areas of xyr1 bypass the need for inducing carbon source, allowing high (hemi)cellulase production even in the presence of glucose. These mutations also change the profile of produced proteins, shifting it more towards xylanase production, and increase the overall protein production in inducing conditions. However, how these mutations alter the metabolism and other cellular processes to cause these changes remains unclear.ResultsIn this study, we aimed to explore changes caused by a point mutation in xyr1 on transcriptomic and metabolic level to better understand the reasons behind the increased protein production in both repressing glucose and inducing lactose conditions. As expected, the expression of many carbohydrate-active enzyme (CAZy) genes was increased in the xyr1 mutant in both conditions. However, their induction was higher under inducing conditions. The xyr1 mutant strain built more biomass and produced more extracellular proteins during growth on lactose compared to the wild type xyr1 strain. Genes involved in oxidoreductive D-galactose catabolism pathway were upregulated in the xyr1 mutant strain, potentially contributing to the more efficient utilization of lactose. In addition to CAZy genes, clustering and enrichment analysis showed over-representation of mitochondria-related Gene Ontology terms in clusters where gene expression was higher in the xyr1 mutant, indicating that mitochondria play a role in the altered metabolic state associated with the xyr1 mutation. Metabolomics revealed that free tyrosine was more abundant in the xyr1 mutant strain in all measured timepoints, whereas multiple fatty acids were less abundant in the mutant strain on glucose.ConclusionsThe results contribute to more in-depth knowledge on T. reesei physiology growing under inducing and repressing carbon sources and gives new insights on the function of the master regulator XYR1. The vast data generated serve as a source for new targets for improved protein production.

Amylase and Cellulase Production from Newly Isolated Bacillus subtilis Using Acid Treated Potato Peel Waste.

Species belonging to the genus Bacillus produce many advantageous extracellular enzymes that have tremendous applications on a commercial scale for the textile, detergent, feed, food, and beverage industries. This study aimed to isolate potent thermo-tolerant amylolytic and cellulolytic bacterium from the local environment. Using the Box-Behnken design of response surface methodology, we further optimized the amylase and cellulase activity. The isolate was identified by 16S rRNA gene sequencing as Bacillus subtilis QY4. This study utilized potato peel waste (PPW) as the biomaterial, which is excessively being dumped in an open environment. Nutritional status of the dried PPW was determined by proximate analysis. All experimental runs were carried out in 250 mL Erlenmeyer flasks containing acid treated PPW as a substrate by the thermos-tolerant Bacillus subtilis QY4 incubated at 37 °C for 72 h of submerged fermentation. Results revealed that the dilute H2SO4 assisted autoclaved treatment favored more amylase production (0.601 IU/mL/min) compared to the acid treatment whereas high cellulase production (1.269 IU/mL/min) was observed in the dilute acid treatment and was found to be very effective compared to the acid assisted autoclaved treatment. The p-value, F-value, and coefficient of determination proved the significance of the model. These results suggest that PPW could be sustainably used to produce enzymes, which offer tremendous applications in various industrial arrays, particularly in biofuel production.

Isolation of an Acidophilic Cellulolytic Bacterial Strain and Its Cellulase Production Characteristics

The aim of the study was to isolate and identify a highly efficient cellulolytic bacteria strain that can be used in acidic environments, and then investigate its cellulase production characteristics for the effective utilization of agricultural waste. For this purpose, we set a series of isolation and screening steps, 21 strains were isolated from soil, and an acidophilic strain labeled as B13-2 with high cellulase production was screened using the Gram’ iodine method and cellulase activity assay; it was identified as Raoultella terrigena. Lastly, the culture conditions such as incubation time, incubation temperature, pH, carbon sources, nitrogen sources, and inoculum size were optimized via single-factor experiments, and on this basis, the cellulase production of strain B13-2 was optimized using response surface methodology with cellulase activity as the optimization goal. The results of the response surface optimization showed that the optimum incubation time is 3.1 days, the optimum temperature is 29.9 °C, the optimum pH is 4.1, and the optimum inoculum size is 1.50%, the cellulase activity reached a maximum of 13.503 U/mL, which was about 140% higher than that before optimization. In particular, strain B13-2 had higher cellulase production when rice straws were used as the natural carbon source. Meanwhile, the SEM pictures demonstrated that the surface of the substrate rice straws in an acidic buffer with strain B13-2 was uneven, with larger holes than in the neutral buffer after incubation. It further proved that this strain has a stronger ability to degrade cellulose under acidic conditions. The B13-2 is a kind of acidophilic cellulolytic bacteria. Therefore, it has the potential to be developed into a silage additive agent and provides a high-quality strain resource for the high-value biotransformation of agricultural waste and lays a certain foundation for the sustainable development of agricultural cultivation.

On-site enriched production of cellulase enzyme using rice straw waste and its hydrolytic performance evaluation through systematic dynamic modeling.

The application of on-site produced cellulolytic enzymes in place of commercial enzymes towards hydrolytic preparations of reducing sugars using inexpensive lignocellulosic wastes is considered the most efficient strategy to accomplish a cost-effective biofuel production process. Along with improved production, intrinsic and systematic performance evaluation of the produced enzyme during the hydrolysis process through kinetic intervention remains a crucial requirement for achieving the improved performance of the process. With this motivation, the present study primarily deals with the nutritionally optimized production strategy of cellulases from rice straw (RS) waste using Trichoderma reesei (MTCC 164). The highest cellulase production was obtained 8.09 ± 0.32g/l in batch mode at optimized combinations of 3.5% (w/v) RS inducer, 3.0% (w/v) lactose, and 1.5% (w/v) peptone. Production was further improved through pH-regulated (pH 5.5 to 6.5) fed-batch fermentations. The enzyme produced at pH 6 was considered for hydrolysis studies at 4 to 10% (w/w) solid loading due to reasonable exoglucanase, endoglucanase, and maximum β-glucosidase activity levels of 9.3 U/ml, 3.87 U/ml, and 2.65 U/ml respectively. Multi-reaction systematic kinetic modeling was implemented to evaluate enzyme performance during hydrolysis, and the values of inhibitory kinetic parameters (K2r = 7.1 < K1r = 18.5 < K3r = 276.6) suggested that sequential conversion of cellulose to glucose by existing enzyme components was more dominant over direct conversion.

A novel fusion transcription factor drives high cellulase and xylanase production on glucose in Trichoderma reesei

To reduce the high cost of (hemi)cellulase production in lignocellulose biorefining, it is important to develop strategies to enhance enzyme productivity from economic and also readily manipulatable carbon sources. In this study, an artificial transcription factor XT was designed by fusing the DNA binding domain of Xyr1 to the transactivation domain of Tmac1. When overexpressed in Trichoderma reesei QM9414 Δxyr1, the XT recombinant strain (OEXT) greatly improved (hemi)cellulase production on repressing glucose compared with QM9414 on Avicel with 1.7- and 8.2-fold increases in pNPCase and xylanase activity, respectively. Both activities were even higher (0.9- and 33.8-fold higher, respectively) than the recombinant strain similarly overexpressing Xyr1. The dramatically enhanced xylanase activities in OEXT resulted from the elevated expression of various hemicellulases in the secretome. Moreover, the enzyme cocktail from OEXT improved the saccharification efficiency toward corn stover by 60% compared with enzymes from QM9414 with equal volume loading.

Effects of different parameters on cellulase production by Trichoderma harzianum TF2 using solid‐state fermentation (SSF)

Solid‐state fermentation is one of the easiest and cheapest methods for producing microbial bioactive com‐ pounds. Trichoderma harzianum has long been recognised as one of the potential fungi for this purpose. Trichoderma sp. were isolated from banana rhizosphere using the soil dilution method and later screened for their ability to produce cellulases using filter paper activity (FPase) and the carboxylmethyl cellulase (CMCase) test. Trichoderma sp. were also subjected to one factor change at a time to determine the effects of different parameters on cellulase production. It was observed that T. harzianum TF2 showed the ability to produce higher cellulase activity when wheat bran was used as the substrate. The results showed that 38.5 U/g of cellulase was produced with the use of wheat bran coupled with an incubation temperature of 28 °C and moisture content of 60%. T. harzianum TF2 showed good potential for use as a culture for cellulase production in this study due to its higher cellulase production under solid‐state fermentation, with the possibility of its application to industry.

Saccharification of pre-treated wheat straw via optimized enzymatic production using Aspergillus niger: Chemical analysis of lignocellulosic matrix

Bioconversion of lignocellulosic wastes to valuable end-products via multi-enzymatic hydrolysis is a potential low costing process for large-scale application in many industrial sectors. Thus, in this study, among thirty local fungal isolates, Aspergillus niger which gave the highest cellulase production, was identified under the accession number MZ062603 in GeneBank. Three types of pre-treatments (0.0–0.7%) acid, (0.0–2%) alkali, and (70–90 °C) hot water were applied to increase wheat straw (WS) digestibility by A. niger, and 1% NaOH treated WS was superior to the other pre-treatments (acid and hot water). During solid-state fermentation, the total cellulolytic activity of [filter-paper cellulase (FPase), carboxy-methyl cellulase (CMCase), and β- glucosidase (βGase)] increased about 2.8-fold. While reducing sugar was increased by 3.1 times. The optimum values of total cellulases activities and reducing sugar (8907.2 and 92.4 mg/gds) were obtained after 3 days of incubation at 30 °C and pH 5.2 at 75% v/w moisture using 3 days old inoculum (106 spores/mL/gds). The WS substrate which was subjected to alkali pre-treatment subsequent to fungal bioconversion was varied in its chemical composition and detailed structure compared to the raw and alkali pre-treatment ones as indicated by its chemical analysis, Scanning electron microscopy (SEM) observation, Fourier Transform Infra-red Spectroscopy (FTIR), and X-Ray Diffraction (XRD). All these analyses revealed that the lignocellulosic matrix was completely destroyed after the fungal treatment.

Statistical optimization of cellulase production by halophilic strains isolated from Tunisian Sebkhas using the renewable marine biomass waste, Posidonia oceanica, as a cellulosic substrate

Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient technology to combat the problem of depleting fossil fuels. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus, main objective of the present study was to optimize composition of culture parameters for enhancing cellulase production by two halophilic strains Oceanobacillus picture E50 and Bacillus vallismortis J77 using Posidonia oceanica waste (POW) as a substrate. A Plackett-Burman design was conducted to study and evaluate the weight of eight cultural variables on bacterial cellulase production. The optimal culture conditions for maximum enzyme yield were developed by maintaining the variables of POW concentration, NaCl concentration, POW liquor, bacterial strain and inoculum size at their higher levels, while keeping substrate particle size, glucose concentration and incubation time at their lower levels. The highest cellulase production in terms of CMCase activity was 950 U/L following this optimization process. Thus, the results obtained in this study demonstrated the potential of utilizing POW as a substrate for enhanced cellulase production by halophilic bacteria. B. vallismortis J77 produced the highest cellulase activity and therefore could be of great interest for saccharification of Posidonia oceanica biomass for biofuel and biorefinery applications.

Extraction and characterization of cellulase from forest and compost soil fungal isolates for the application of straw degradation

Cellulases are complex hydrolytic enzymes working synergistically on the hydrolysis of cellulolytic materials for the production of simple sugars. These enzymes have tremendous environmental, industrial and agricultural applications including enhancement of the degradability of lignocellulosic materials for cattle’s feed. The aim of this study was to isolate fungus and extract cellulose enzyme from forest and compost soil samples and examine the extent to which these enzymes enhance the degradability of finger millet and oat straw for making palatable cattle’s feed. Accordingly, a total of 53 fungal isolates were isolated from forest and compost soil. On 1% CMC media resulted 40% were cellulolytic fungal species. Six were selected based on their clear zone. These isolates belonged to the genera Trichoderma, Aspergillus and Penicillium based on their morphological characteristics. Cultivation of fungal isolates for cellulase production using submerged fermentation and Solid-state fermentation was undertaken through make the variation on their levels of different growth conditions such as temperature, pH, minerals and substrates. The results showed that the highest cellulase production i.e CMCase 83.12 ± 3.18 U/dL and FPase 44.51± 0.391FPU/dL were obtained from FSI6 in FMS supplemented SSF at 280C and a pH of 6.

Response-Surface Statistical Optimization of Submerged Fermentation for Pectinase and Cellulase Production by Mucor circinelloides and M. hiemalis

Cellulase and pectinase are degrading cellulosic and pectic substances that form plant cell walls and, thereby, they have a wide range of applications in the agro-industrial by-products recycling and food industries. In the current research, Mucor circinelloides and M. hiemalis strains were tested for their ability to produce cellulase and pectinase from tangerine peel by submerged fermentation. Experiments on five variables: temperature, pH, incubation period, inoculum size, and substrate concentration, were designed with a Box–Behnken design, as well as response surface methodology (RSM), and analysis of variance was performed. In addition, cellulase and pectinase were partially purified and characterized. At their optimum parameters, M. circinelloides and M. hiemalis afforded high cellulase production (37.20 U/mL and 33.82 U/mL, respectively) and pectinase (38.02 U/mL and 39.76 U/mL, respectively). The partial purification of M. circinelloides and M. hiemalis cellulase produced 1.73- and 2.03-fold purification with 31.12 and 32.02% recovery, respectively; meanwhile, 1.74- and 1.99-fold purification with 31.26 and 31.51% recovery, respectively, were obtained for pectinase. Partially purified cellulase and pectinase from M. circinelloides and M. hiemalis demonstrated the highest activity at neutral pH, and 70 and 50 °C, for cellulase and 50 and 60 °C, for pectinase, respectively. Moreover, 10 mM of K+ increased M. circinelloides enzymatic activity. The production of cellulase and pectinase from M. circinelloides and M. hiemalis utilizing RSM is deemed profitable for the decomposition of agro-industrial wastes.

Screening and characterization of a low-temperature-resistant cellulose-degrading strain, Trichoderma harzianum L-8, from a primitive forest

Low temperature is a major factor limiting the bio-sustainable and efficient conversion of cellulose-based resources in cold regions. In this study, a low-temperature resistant cellulose-degrading fungus with high cellulase production was screened from samples found in a primitive forest in Daqing by straw using the enrichment-restricted culture technique. The fungus was identified as genus Trichoderma harzianum, strain L-8 by morphological and molecular biological analysis. The enzyme production conditions were optimized via response surface methodology, and the optimal conditions for the enzyme production of Trichoderma harzianum L-8 were as follows: a CMC-Na addition of 10.63 g·L-1, an ammonium sulfate addition of 2.22 g·L-1, an initial pH of 5.29, and a lecithin addition of 5.18 g·L-1 when the CMCase reached 53.40 IU·mL-1. The leading enzyme families of Trichoderma harzianum L-8 were identified via proteomic analysis. Proteases including glycosyl hydrolase family 3-4 and cellobiohydrolase play important roles in cellulose degradation. The strain Trichoderma harzianum L-8 showed a strong cellulose degradation ability under low temperatures, providing strain resources for cellulose resource biotransformation technology in cold regions.

Screening of cellulase-producing bacteria and their effect on the chemical composition and aroma quality improvement of cigar wrapper leaves

A high yield cellulase production strain was screened from the surface of cigar wrapper leaves and added to cigar wrapper leaves for fermentation in the present study. The enzyme-producing strain was identified as Bacillus velezensis, and its maximum carboxymethyl cellulase activity could reach 44.3 U/mL. Making the cellulose degradation rate of cigar wrapper leaves as the index, the fermentation temperature, inoculum concentration, and carbon and nitrogen source addition concentration were optimized by response surface methodology. The results showed that under the conditions of inoculation concentration of 20%, fermentation temperature of 37 °C, addition of 4.5 ‰ glucose and 2.25 ‰ glutamate, the highest degradation rate of tobacco cellulose was 31.7%. Significance analysis results showed that the order of factors affecting the degradation rate of cigar wrapper cellulose was: inoculation concentration &gt; carbon and nitrogen source addition concentration &gt; temperature. The analysis of aroma substances under the optimal conditions found that the total amount of aroma substances of cigar wrapper increased by 26.1% compared with that before fermentation, in which β-damascenone, megastigmatrienone, farnesyl acetone, and solanone were significantly improved. The results were conducive to improving the aroma quality and concentration of cigar wrapper leaves.

Screening of highly efficient fungi for the degradation of lignocelluloses by ionic liquids-assisted cellulase

A successful conversion of lignocellulosic biomass into biofuel could be achieved through an ionic liquids (ILs)-pretreatment and an enzyme saccharification. In this study, the fungal strains with high cellulase productions were isolated and identified as Penicillium janthinellum FHH1 and P. oxalicum FLY4. A high cellulase production was obtained at pH 4.0 and 30 °C for P. janthinellum FHH1 with corn stalk and beef extract for 9 days and for P. janthinellum FHH1 with corn stalk and peptone for 7 days. A pH range between 4.0 to 6.0 and a temperature range from 55 °C to 60 °C were applicable for cellulase with high activities during hydrolysis. A high NaCl-stability and a relatively high ILs-stability were found for cellulase obtained from P. oxalicum FLY4. High yields of reducing sugar were obtained by enzymatic hydrolysis of the original corn stalk and ILs-pretreated corn stalk. P. oxalicum FLY4 has a great potential for ILs-assisted fungi in the degradation of lignocellulosic biomass into value-added products.

CRISPR/Cas9-mediated genome editing directed by a 5S rRNA\u2013tRNAGly hybrid promoter in the thermophilic filamentous fungus Humicola insolens

BackgroundHumicola insolens is a filamentous fungus with high potential of producing neutral and heat- and alkali-resistant cellulase. However, the genetic engineering tools, particularly the genome-editing tool, are scarce, hindering the study of cellulase expression regulation in this organism.ResultsHerein, a CRISPR/Cas9 genome-editing system was established in H. insolens based on a hybrid 5S rRNA–tRNAGly promoter. This system is superior to the HDV (hepatitis delta virus) system in genome editing, allowing highly efficient single gene destruction in H. insolens with rates of deletion up to 84.1% (37/44). With this system, a putative pigment synthesis gene pks and the transcription factor xyr1 gene were disrupted with high efficiency. Moreover, the extracellular protein concentration and cellulase activity largely decreased when xyr1 was deleted, demonstrating for the first time that Xyr1 plays an important role in cellulase expression regulation.ConclusionsThe established CRISPR/Cas9 system is a powerful genetic operation tool for H. insolens, which will accelerate studies on the regulation mechanism of cellulase expression and engineering of H. insolens for higher cellulase production.

Lignocelluloytic activities and composition of bacterial community in the camel rumen

The camel is well-adapted to utilize the poor-quality forages in the harsh desert conditions as the camel rumen sustains fibrolytic microorganisms, mainly bacteria that are capable of breaking down the lignocellulosic biomass efficiently. Exploring the composition of the bacterial community in the rumen of the camel and quantifying their cellulolytic and xylanolytic activities could lead to understanding and improving fiber fermentation and discovering novel sources of cellulases and xylanases. In this study, Illumina MiSeq sequencing of the V4 region on 16S rRNA was applied to identify the bacterial and archaeal communities in the rumen of three camels fed wheat straw and broom corn. Furthermore, rumen samples were inoculated into bacterial media enriched with xylan and different cellulose sources, including filter paper (FP), wheat straw (WS), and alfalfa hay (AH) to assess the ability of rumen bacteria to produce endo-cellulase and endo-xylanase at different fermentation intervals. The results revealed that the phylum Bacteroidetes dominated the bacterial community and Candidatus Methanomethylophilus dominated the archaeal community. Also, most of the bacterial community has fibrolytic potential and the dominant bacterial genera were Prevotella, RC9_gut_group, Butyrivibrio, Ruminococcus, Fibrobacteres, and Treponema. The highest xylanase production (884.8 mU/mL) was observed at 7 days. The highest cellulase production (1049.5 mU/mL) was observed when rumen samples were incubated with Alfalfa hay for 7 days.

Mutagenesis of Aspergillus aculeatus by 60Co-γ irradiation for high production of potential ILs-tolerant cellulase

Ionic liquids (ILs) are effective solvents for lignocellulose pretreatment. Enzymatic saccharification converts pretreated lignocelluloses into valuable products, and IL-tolerant cellulase improves the enzymatic efficiency and the reuse of ILs. In this study, a fungal strain with a relatively high cellulase production was isolated and identified as Aspergillus aculeatus G1-3. The high production of β-glucosidase (1.943 U per mL), CMCase (1.303 U per mL), and FPase (0.165 U per mL) was obtained using corn stover as the carbon source and peptone as the nitrogen source. The results were obtained at pH 8.0 and 30 °C with an inoculation size of 3% (volume per volume) for 7 days. A mutant strain Aspergillus aculeatus P6 with β-glucosidase (7.023 U per mL), CMCase (1.543 U per mL), and FPase (0.098 U per mL) was obtained by 60Co-γ irradiation. The cellulase activity was measured at pH 5.0 and 60 °C for enzymatic hydrolysis. The cellulase from mutant strains was stable in different concentrations of 1-ethyl-3-methylimidazolium acetate. Enzymatic saccharification of the original corn stover and ILs-pretreated corn stover was successfully performed with high sugar yields. The mutant strains of Aspergillus aculeatus have great potential for their further application in the conversion of lignocellulosic biomass into biofuels.

Acanthophysium sp. KMF001, a New Strain with High Cellulase Activity

Cellulase is an eco-friendly biocatalyst, and its demand is growing in many industrial applications such as food, textile, paper, and bioenergy. Strains with a high cellulase activities are the starting point for the economic production of cellulase. In a previous study, Acanthophysium sp. KMF001 with high cellulase production ability was selected among 54 wood-rotting fungi. In this study, we evaluated the cellulase productivity of Acanthophysium sp. KMF001 quantitatively and analyzed its taxonomic location using a genetic method. Acanthophysium sp. KMF001 showed high cellulase productivity similar to that of Acanthophysium bisporum and was much better than A. bisporum in specific enzyme activity. The 28S rRNA sequence of Acanthophysium sp. KMF001 was similar to that of Acanthophysium lividocaeruleum MB1825, with 98.40% homology. Phylogenetic analysis suggested that Acanthophysium sp. KMF001 is a new strain. In this study, we propose a new strain with high cellulase productivity.

High-yield cellulase and LiP production after SSF of agricultural wastes by Pleurotus ostreatus using different surfactants

Abstract White rot fungi are well known for their ability to degrade the structures of the plant cell walls and to produce of a wide range of extracellular enzymes using lignocellulosic biomass as substrate. In an attempt to reduce the costs and obtain high-yield of lignocellulolytic enzymes such as cellulases and lignin peroxidase, their production has been studied by solid-state fermentation using low-cost agroindustrial residues as carbon sources and surfactants. In this context, this work aimed the produce cellulases and lignin peroxidase from the fungus Pleurotus ostreatus PLO 9 using banana pseudostem, jatropha and coconut fiber as the substrates supplemented with the surfactants tween 80, sodium dodecyl sulfate and glycerol. The results showed the maximum lignin peroxidase activity was obtained after 15 days of fermentation using jatropha supplemented with sodium dodecyl sulfate (49916 ± 541 U g−1 of substrate). The highest cellulase production was reached after 5 days of fermentation using banana pseudostem supplemented with glycerol as the substrate (19.5 ± 0.2 FPU g−1 of substrate). The results indicate that it was possible to reach significant activities of cellulase and LiP using the white rot Pleurotus ostreatus after solid state fermentation of lignocellulosic biomass using surfactants as exogenous inducers.

A novel Trichoderma reesei mutant RP698 with enhanced cellulase production

A new strain of Trichoderma reesei (teleomorph Hypocrea jecorina) with high cellulase production was obtained by exposing the spores from T. reesei QM9414 to an ultraviolet light followed by selecting fast-growing colonies on plates containing CMC (1% w/v) as the carbon source. The mutant T. reesei RP698 reduced cultivation period to 5 days and increased tolerance to the end-products of enzymatic cellulose digestion. Under submerged fermentation conditions, FPase, CMCase, and Avicelase production increased up to 2-fold as compared to the original QM9414 strain. The highest levels of cellulase activity were obtained at 27 °C after 72 h with Avicel®, cellobiose, and sugarcane bagasse as carbon sources. The temperature and pH activity optima of the FPase, CMCase, and Avicelase were approximately 60 °C and 5.0, respectively. The cellulase activity was unaffected by the addition of 140 mM glucose in the enzyme assay. When T. reesei RP698 crude extract was supplemented by the addition of β-glucosidase from Scytalidium thermophilum, a 2.3-fold increase in glucose release was observed, confirming the low inhibition by the end-product of cellulose hydrolysis. These features indicate the utility of this mutant strain in the production of enzymatic cocktails for biomass degradation.