Enhancing soil quality and yield through microbial assisted in-situ residue management in rice-rice cropping system in Odisha, Eastern India.

Crop residue management has become more challenging with intensive agricultural operations. Zero tillage and crop residue returns, along with the enhancement of in-situ residue decomposition through microbial intervention, are essential measures for preserving and enhancing soil quality. To address this problem in view of stubble burning, field experiments were conducted in rice-rice (variety Swarna) cropping systems under lowland conditions, wherein the following different residue management practices were adopted viz., conventional cultivation (CC), residue incorporation (RI @ 6 t paddy straw ha-1), residue retention (RR @6 t paddy straw ha-1), and zero tillage (ZT). In this experiment, two microbial products i.e. solid microbial consortium (SMC) at 2.0 kg ha-1) and capsule (10 numbers ha-1), were evaluated in both Rabi (dry) and Kharif (wet) seasons under different residue management practices. The results on soil microbial properties showed that application of either SMC or capsule based formulation could significantly improve the soil organic carbon (SOC) content in ZT (9.51 g/kg), followed by RI (9.36 g/kg), and RR (9.34 g/kg) as compared to CC (7.61 g/kg). There were significant differences in the soil functional properties (AcP, AkP, FDA, and DHA) with microbial interventions across all residue management practices. SOC was significantly positive correlated with cellulase (R2 = 0.64, p < 0.001), ß-glucosidase (R2 = 0.61, p < 0.001), and laccase (R2 = 0.66, p < 0.001) activity; however, the regression coefficients varied significantly with microbial intervention. Moreover, the availability of N, P, and K in soil was significantly (p < 0.05) improved under microbial treatments with either RR or RI practices. Among the different methods of residues management practices, RI with microbial intervention registered a consistent yield improvement (8.4-17.8%) compared to conventional practices with microbial intervention. The present findings prove that the application of decomposing microbial consortia for in-situ rice residue management under field conditions significantly enhances soil quality and crop yield compared to conventional practices.

Enzymatic machinery of wood-inhabiting fungi that degrade temperate tree species.

Deadwood provides habitat for fungi and serves diverse ecological functions in forests. We already have profound knowledge of fungal assembly processes, physiological and enzymatic activities, and resulting physico-chemical changes during deadwood decay. However, in situ detection and identification methods, fungal origins, and a mechanistic understanding of the main lignocellulolytic enzymes are lacking. This study used metaproteomics to detect the main extracellular lignocellulolytic enzymes in 12 tree species in a temperate forest that have decomposed for 8 ½ years. Mainly white-rot (and few brown-rot) Basidiomycota were identified as the main wood decomposers, with Armillaria as the dominant genus; additionally, several soft-rot xylariaceous Ascomycota were identified. The key enzymes involved in lignocellulolysis included manganese peroxidase, peroxide-producing alcohol oxidases, laccase, diverse glycoside hydrolases (cellulase, glucosidase, xylanase), esterases, and lytic polysaccharide monooxygenases. The fungal community and enzyme composition differed among the 12 tree species. Ascomycota species were more prevalent in angiosperm logs than in gymnosperm logs. Regarding lignocellulolysis as a function, the extracellular enzyme toolbox acted simultaneously and was interrelated (e.g. peroxidases and peroxide-producing enzymes were strongly correlated), highly functionally redundant, and present in all logs. In summary, our in situ study provides comprehensive and detailed insight into the enzymatic machinery of wood-inhabiting fungi in temperate tree species. These findings will allow us to relate changes in environmental factors to lignocellulolysis as an ecosystem function in the future.

Combined hemicellulolytic and phenoloxidase activities of Thermobacillus xylanilyticus enable growth on lignin-rich substrates and the release of phenolic molecules.

Major challenge in biorefineries is the use of all lignocellulosic components, particularly lignins. In this study, Thermobacillus xylanilyliticus grew on kraft lignin, steam-exploded and native wheat straws produced different sets of phenoloxidases and xylanases, according to the substrate. After growth, limited lignin structural modifications, mainly accompanied by a decrease in phenolic acids was observed by Nuclear Magnetic Resonance spectroscopy. The depletion of p-coumaric acid, vanillin and p-hydroxybenzaldehyde combined to vanillin production in the culture media indicated that the bacterium can transform some phenolic compounds. Proteomic approaches allowed the identification of 29 to 33 different hemicellulases according to the substrates. Twenty oxidoreductases were differentially expressed between kraft lignin and steam-exploded wheat straw. These oxidoreductases may be involved in lignin and aromatic compound utilization and detoxification. This study highlights the potential value of Thermobacillus xylanilyticus and its enzymes in the simultaneous valorization of hemicellulose and phenolic compounds from lignocelluloses.

Rice Husk, Brewer's Spent Grain, and Vine Shoot Trimmings as Raw Materials for Sustainable Enzyme Production.

Solid by-products with lignocellulosic structures are considered appropriate substrates for solid-state fermentation (SSF) to produce enzymes with diverse industrial applications. In this work, brewer's spent grain (BSG), rice husk (RH), and vine shoot trimmings (VSTs) were employed as substrates in SSF with Aspergillus niger CECT 2088 to produce cellulases, xylanases, and amylases. The addition of 2% (NH4)2SO4 and 1% K2HPO4 to by-products had a positive effect on enzyme production. Substrate particle size influenced enzyme activity and the overall highest activities were achieved at the largest particle size (10 mm) of BSG and RH and a size of 4 mm for VSTs. Optimal substrate composition was predicted using a simplex centroid mixture design. The highest activities were obtained using 100% BSG for ß-glucosidase (363 U/g) and endo-1,4-ß-glucanase (189 U/g), 87% BSG and 13% RH for xylanase (627 U/g), and 72% BSG and 28% RH for amylase (263 U/g). Besides the optimal values found, mixtures of BSG with RH or VSTs proved to be alternative substrates to BSG alone. These findings demonstrate that SSF bioprocessing of BSG individually or in mixtures with RH and VSTs is an efficient and sustainable strategy to produce enzymes of significant industrial interest within the circular economy guidelines.

Transcriptomic insights into the roles of the transcription factors Clr1, Clr2 and Clr4 in lignocellulose degradation of the thermophilic fungal platform Thermothelomyces thermophilus.

Introduction: Thermothelomyces thermophilus, formerly known as Myceliophthora thermophila, is used in industry to produce lignocellulolytic enzymes and heterologous proteins. However, the transcriptional network driving the expression of these proteins remains elusive. As a first step to systematically uncover this network, we investigated growth, protein secretion, and transcriptomic fingerprints of strains deficient in the cellulolytic transcriptional regulators Clr1, Clr2, and Clr4, respectively. Methods: The genes encoding Clr1, Clr2, and Clr4 were individually deleted using split marker or the CRISPR/Cas12a technology and the resulting strains as well as the parental strain were cultivated in bioreactors under chemostat conditions using glucose as the carbon source. During steady state conditions, cellulose was added instead of glucose to study the genetic and cellular responses in all four strains to the shift in carbon source availability. Results: Notably, the clr1 and clr2 deletion strains were unable to continue to grow on cellulose, demonstrating a key role of both regulators in cellulose catabolism. Their transcriptomic fingerprints uncovered not only a lack of cellulase gene expression but also reduced expression of genes predicted to encode hemicellulases, pectinases, and esterases. In contrast, the growth of the clr4 deletion strain was very similar compared to the parental strain. However, a much stronger expression of cellulases, hemicellulases, pectinases, and esterases was observed. Discussion: The data gained in this study suggest that both transcriptional regulators Clr1 and Clr2 activate the expression of genes predicted to encode cellulases as well as hemicellulases, pectinases, and esterases. They further suggest that Clr1 controls the basal expression of cellulases and initiates the main lignocellulolytic response to cellulose via induction of clr2 expression. In contrast, Clr4 seems to act as a repressor of the lignocellulolytic response presumably via controlling clr2 expression. Comparative transcriptomics in all four strains revealed potentially new regulators in carbohydrate catabolism and lignocellulolytic enzyme expression that define a candidate gene list for future analyses.

Nitrogen addition increases mass loss of gymnosperm but not of angiosperm deadwood without changing microbial communities.

Enhanced nitrogen (N) deposition due to combustion of fossil fuels and agricultural fertilization is a global phenomenon which has severely altered carbon (C) and N cycling in temperate forest ecosystems in the northern hemisphere. Although deadwood holds a substantial amount of C in forest ecosystems and thus plays a crucial role in nutrient cycling, the effect of increased N deposition on microbial processes and communities, wood chemical traits and deadwood mass loss remains unclear. Here, we simulated high N deposition rates by adding reactive N in form of ammonium-nitrate (40 kg N ha-1 yr-1) to deadwood of 13 temperate tree species over nine years in a field experiment in Germany. Non-treated deadwood from the same logs served as control with background N deposition. Our results show that chronically elevated N levels alters deadwood mass loss alongside respiration, enzymatic activities and wood chemistry depending on tree clade and species. In gymnosperm deadwood, elevated N increased mass loss by +38 %, respiration by +37 % and increased laccase activity 12-fold alongside increases of white-rot fungal abundance +89 % (p = 0.03). Furthermore, we observed marginally significant (p = 0.06) shifts of bacterial communities in gymnosperm deadwood. In angiosperm deadwood, we did not detect consistent effects on mass loss, physico-chemical properties, extracellular enzymatic activity or changes in microbial communities except for changes in abundance of 10 fungal OTUs in seven tree species and 28 bacterial OTUs in 10 tree species. We conclude that N deposition alters decomposition processes exclusively in N limited gymnosperm deadwood in the long term by enhancing fungal activity as expressed by increases in respiration rate and extracellular enzyme activity with minor shifts in decomposing microbial communities. By contrast, deadwood of angiosperm tree species had higher N concentrations and mass loss as well as community composition did not respond to N addition.

Papermaking wastewater treatment coupled to 2,3-butanediol production by engineered psychrotrophic Raoultella terrigena.

The simultaneous bioremediation and bioconversion of papermaking wastewater by psychrotrophic microorganisms holds great promise for developing sustainable environments and economies in cold regions. Here, the psychrotrophic bacterium Raoultella terrigena HC6 presented high endoglucanase (26.3 U/mL), xylosidase (732 U/mL), and laccase (8.07 U/mL) activities for lignocellulose deconstruction at 15 °C. mRNA monitoring and phenotypic variation analyses confirmed that cold-inducible cold shock protein A (CspA) facilitated the expression of the cel208, xynB68, and lac432 genes to increase the enzyme activities in strain HC6. Furthermore, the cspA gene-overexpressing mutant (strain HC6-cspA) was deployed in actual papermaking wastewater and achieved 44.3%, 34.1%, 18.4%, 80.2% and 100% removal rates for cellulose, hemicellulose, lignin, COD, and NO3--N at 15 °C. Simultaneously, 2,3-butanediol (2,3-BD) was produced from the effluent with a titer of 2.98 g/L and productivity of 0.154 g/L/h. This study reveals an association between the cold regulon and lignocellulolytic enzymes and provides a promising candidate for simultaneous papermaking wastewater treatment and 2,3-BD production.

New insight into the bark beetle ips typographus bacteriome reveals unexplored diversity potentially beneficial to the host.

BACKGROUND: Ips typographus (European spruce bark beetle) is the most destructive pest of spruce forests in Europe. As for other animals, it has been proposed that the microbiome plays important roles in the biology of bark beetles. About the bacteriome, there still are many uncertainties regarding the taxonomical composition, insect-bacteriome interactions, and their potential roles in the beetle ecology. Here, we aim to deep into the ecological functions and taxonomical composition of I. typographus associated bacteria. RESULTS: We assessed the metabolic potential of a collection of isolates obtained from different life stages of I. typographus beetles. All strains showed the capacity to hydrolyse one or more complex polysaccharides into simpler molecules, which may provide an additional carbon source to its host. Also, 83.9% of the strains isolated showed antagonistic effect against one or more entomopathogenic fungi, which could assist the beetle in its fight against this pathogenic threat. Using culture-dependent and -independent techniques, we present a taxonomical analysis of the bacteriome associated with the I. typographus beetle during its different life stages. We have observed an evolution of its bacteriome, which is diverse at the larval phase, substantially diminished in pupae, greater in the teneral adult phase, and similar to that of the larval stage in mature adults. Our results suggest that taxa belonging to the Erwiniaceae family, and the Pseudoxanthomonas and Pseudomonas genera, as well as an undescribed genus within the Enterobactereaceae family, are part of the core microbiome and may perform vital roles in maintaining beetle fitness. CONCLUSION: Our results indicate that isolates within the bacteriome of I. typographus beetle have the metabolic potential to increase beetle fitness by proving additional and assimilable carbon sources for the beetle, and by antagonizing fungi entomopathogens. Furthermore, we observed that isolates from adult beetles are more likely to have these capacities but those obtained from larvae showed strongest antifungal activity. Our taxonomical analysis showed that Erwinia typographi, Pseudomonas bohemica, and Pseudomonas typographi species along with Pseudoxanthomonas genus, and putative new taxa belonging to the Erwiniaceae and Enterobacterales group are repeatedly present within the bacteriome of I. typographus beetles, indicating that these species might be part of the core microbiome. In addition to Pseudomonas and Erwinia group, Staphylococcus, Acinetobacter, Curtobacterium, Streptomyces, and Bacillus genera seem to also have interesting metabolic capacities but are present in a lower frequency. Future studies involving bacterial-insect interactions or analysing other potential roles would provide more insights into the bacteriome capacity to be beneficial to the beetle.

Comparative assessment on lignocellulose degrading enzymes and bioethanol production from spent mushroom substrate of Calocybe indica and Volvariella volvacea.

In the current study, we compared the production of extracellular lignocellulose degrading enzymes and bioethanol from the spent mushroom substrate (SMS) of Calocybe indica and Volvariella volvacea. From SMS at different stages of the mushroom development cycle, ligninolytic and hydrolytic enzymes were analysed. The activities of lignin-degrading enzymes, including lignin peroxidase (LiP), laccase, and manganese peroxidase (MnP) were maximal in the spawn run and primordial stages, while hydrolytic enzymes including xylanase, cellobiohydrolase (CBH), and carboxymethyl cellulase (CMCase) showed higher activity during fruiting bodies development and at the end of the mushroom growth cycle. SMS of V. volvacea showed relatively lower ligninase activity than the SMS of C. indica, but had the maximum activity of hydrolytic enzymes. The enzyme was precipitated with acetone and further purified with the DEAE cellulose column. The maximum yield of reducing sugars was obtained after hydrolysis of NaOH (0.5 M) pretreated SMS with a cocktail of partially purified enzymes (50% v/v). After enzymatic hydrolysis, the total reducing sugars were 18.68 ± 0.34 g/l (SMS of C. indica) and 20.02 ± 0.87 g/l (SMS of V. volvacea). We observed the highest fermentation efficiency and ethanol productivity (54.25%, 0.12 g/l h) obtained from SMS hydrolysate of V. volvacea after 48 h at 30 ± 2 °C, using co-culture of Saccharomyces cerevisiae MTCC 11,815 and Pachysolen tannophilus MTCC 1077.

Lignocellulolytic Biocatalysts: The Main Players Involved in Multiple Biotechnological Processes for Biomass Valorization.

Human population growth, industrialization, and globalization have caused several pressures on the planet's natural resources, culminating in the severe climate and environmental crisis which we are facing. Aiming to remedy and mitigate the impact of human activities on the environment, the use of lignocellulolytic enzymes for biofuel production, food, bioremediation, and other various industries, is presented as a more sustainable alternative. These enzymes are characterized as a group of enzymes capable of breaking down lignocellulosic biomass into its different monomer units, making it accessible for bioconversion into various products and applications in the most diverse industries. Among all the organisms that produce lignocellulolytic enzymes, microorganisms are seen as the primary sources for obtaining them. Therefore, this review proposes to discuss the fundamental aspects of the enzymes forming lignocellulolytic systems and the main microorganisms used to obtain them. In addition, different possible industrial applications for these enzymes will be discussed, as well as information about their production modes and considerations about recent advances and future perspectives in research in pursuit of expanding lignocellulolytic enzyme uses at an industrial scale.

Solid-State Fermentation of Steam-Exploded Opuntia ficus-indica Cladodes Using Trichoderma reesei CCT-2768 for the Production of Cellulolytic Enzymes.

The sustainable development of the drylands, i.e., regions with limited availability of water, depends on the exploitation of the few biomass types that can thrive in such conditions, such as the Opuntia ficus-indica, a plant of the Cactaceae family. In the present study, the cladodes of O. ficus-indica were used as a substrate by the fungus Trichoderma reesei CCT-2768 for the production of cellulolytic enzymes through solid-state fermentation. Firstly, the extraction of the mucilage, soluble components of industrial interest, was evaluated. Temperature, water-to-biomass ratio, and time of mixture were varied using an experimental design and impacted, especially, the pectin removal. Then, the lignocellulosic residue was used for the production of enzymes; the effect of the water activity, biomass pretreatment, mineral supplementation, temperature, and inoculum size on the enzymatic production were investigated using two sets of experimental designs. The steam explosion pretreatment exposed the fiber to the microbial action and boosted the enzyme production, provided that the medium was supplemented with salts. This combination has improved the production of xylanase, CMCase, FPase, and polygalacturonase by 27, 62, 98, and 185%, respectively. The temperature of 35 °C was determined as the optimal for the production of FPase, xylanase, and polygalacturonase, while no effect was observed on the production of CMCase and ß-glucosidase. The optimization of the enzymatic production performed in this study can potentially provide a new application for the Opuntia biomass and improve the sustainable development of the drylands.

Genomic Analysis to Elucidate the Lignocellulose Degrading Capability of a New Halophile Robertkochia solimangrovi.

Robertkochia solimangrovi is a proposed marine bacterium isolated from mangrove soil. So far, the study of this bacterium is limited to taxonomy only. In this report, we performed a genomic analysis of R. solimangrovi that revealed its lignocellulose degrading ability. Genome mining of R. solimangrovi revealed a total of 87 lignocellulose degrading enzymes. These enzymes include cellulases (GH3, GH5, GH9 and GH30), xylanases (GH5, GH10, GH43, GH51, GH67, and GH115), mannanases (GH2, GH26, GH27 and GH113) and xyloglucanases (GH2, GH5, GH16, GH29, GH31 and GH95). Most of the lignocellulolytic enzymes encoded in R. solimangrovi were absent in the genome of Robertkochia marina, the closest member from the same genus. Furthermore, current work also demonstrated the ability of R. solimangrovi to produce lignocellulolytic enzymes to deconstruct oil palm empty fruit bunch (EFB), a lignocellulosic waste found abundantly in palm oil industry. The metabolic pathway taken by R. solimangrovi to transport and process the reducing sugars after the action of lignocellulolytic enzymes on EFB was also inferred based on genomic data. Collectively, genomic analysis coupled with experimental studies elucidated R. solimangrovi to serve as a promising candidate in seawater based-biorefinery industry.

Enhancing Antioxidants Extraction from Agro-Industrial By-Products by Enzymatic Treatment.

Nowadays, agro-industrial by-products are of increasing interest as a source of antioxidant compounds. Thus, alternative green techniques to extract antioxidant compounds have been pursued. The use of enzymes to release bioactive compounds through antioxidant activity reduces the environmental impact caused by traditional extraction systems using organic solvents. A crude enzymatic extract containing carbohydrolases was produced by solid-state fermentation (SSF) of an olive pomace and brewery spent-grain combination. The crude extract was evaluated at different temperatures and pH values and its thermostability was studied. Results showed that ß-glucosidase and cellulase were more stable than xylanase, particularly cellulase, which kept 91% of its activity for 72 h at 45 °C. The extract was also applied in enzymatic treatments (ET) to liberate antioxidant compounds from winery, olive mill and brewery by-products under optimal conditions for enzymatic activities. The highest antioxidant activity was found in extracts obtained after enzymatic treatment of exhausted olive pomace (EOP). Enzymatic crude extract produced by SSF was successfully applied in the extraction of antioxidant compounds from winery, olive mill and brewery by-products. Thus, integrating SSF and enzymatic technologies is a valuable approach to implement circular economy practices in the agro-food industry.

In Situ Corn Fiber Conversion for Ethanol Improvement by the Addition of a Novel Lignocellulolytic Enzyme Cocktail.

Corn mashes have high-viscosity and high-sugar characteristics, which hinders yeast-fermentation efficiency and the ethanol yield increase. The excessive viscosity of corn mash is caused by the unutilized cellulose in corn kernel fiber. A novel lignocellulolytic enzymes cocktail with strong substrate specificity was prepared for high-viscosity, high-sugar corn mash. The in situ conversion of corn mashes with novel lignocellulolytic enzymes at the optimum cellulase dosage of 50 FPU/L resulted in about 12% increased ethanol concentration compared with the reference mash at different batch-fermentation scales. Adding the lignocellulolytic enzymes caused the greatest decrease in viscosity of corn mash and residual sugars by 40.9% and 56.3%, respectively. Simultaneously, the application of lignocellulolytic enzymes increased the value of the dried distiller's grain with solubles (DDGS) by increasing the protein content by 5.51%. The in situ conversion of cellulose can decrease the fermentation broth viscosity and improve the rheological property, thereby improving the ethanol yield. With the same amount of material, the application of the novel enzymes cocktail can enhance the ethanol yield by more than 12%. A quarter of the ethanol yield increase was due to the further hydrolysis of starch, while three quarters to cellulose. Thus, this technology will increase the net revenue of bioethanol industrialization.

Mycoremediation of lignocellulosic biorefinery sludge: A reinvigorating approach for organic contaminants remediation with simultaneous production of lignocellulolytic enzyme cocktail.

This research work aims to valorize lignocellulosic biorefinery sludge with genetically engineered Trichoderma atroviride for simultaneous removal of organic contaminants, fermentation inhibitors, and lignocellulolytic enzyme cocktail production. Upon analysis, three phenolic compounds (42.6 ± 3.6 µg/g), two polycyclic aromatic hydrocarbons (0.42 ± 0.06 µg/g) and five fermentation inhibitors (2.5 ± 0.3 mg/g) were detected in the sludge. Bioaugmentation of sludge with 72 h-old T. atroviride (5%) results in the production of cellulase (21 U/g), xylanase (84 U/g), laccase (20 U/g), lignin peroxidase (14 U/g) and aryl alcohol oxidase (116 U/g), along with the concomitant removal of organic contaminants (phenol, 2, 4-dinitrophenol, pentchlorophenol, phenanthrene, benzo(a)pyrene) and fermentation inhibitors (furfural, 5-hydroxymethylfurfural, levulinic acid, ferulic acid, and catechol). Subsequently, the enrichment of sludge with nutrients and rhamnolipids enhanced the enzyme production by 5-6-fold and resulted in the removal of 85-95% of organic contaminants and fermentation inhibitors, which constitutes an eco-friendly process.

Analysis of carbohydrate-active enzymes and sugar transporters in Penicillium echinulatum: A genome-wide comparative study of the fungal lignocellulolytic system.

Penicillium echinulatum 2HH is an ascomycete well known for its production of cellulolytic enzymes. Understanding lignocellulolytic and sugar uptake systems is essential to obtain efficient fungi strains for the production of bioethanol. In this study we performed a genome-wide functional annotation of carbohydrate-active enzymes and sugar transporters involved in the lignocellulolytic system of P. echinulatum 2HH and S1M29 strains (wildtype and mutant, respectively) and eleven related fungi. Additionally, signal peptide and orthology prediction were carried out. We encountered a diverse assortment of cellulolytic enzymes in P. echinulatum, especially in terms of ß-glucosidases and endoglucanases. Other enzymes required for the breakdown of cellulosic biomass were also found, including cellobiohydrolases, lytic cellulose monooxygenases and cellobiose dehydrogenases. The S1M29 mutant, which is known to produce an increased cellulase activity, and the 2HH wild type strain of P. echinulatum did not show significant differences between their enzymatic repertoire. Nevertheless, we unveiled an amino acid substitution for a predicted intracellular ß-glucosidase of the mutant, which might contribute to hyperexpression of cellulases through a cellodextrin induction pathway. Most of the P. echinulatum enzymes presented orthologs in P. oxalicum 114-2, supporting the presence of highly similar cellulolytic mechanisms and a close phylogenetic relationship between these fungi. A phylogenetic analysis of intracellular ß-glucosidases and sugar transporters allowed us to identify several proteins potentially involved in the accumulation of intracellular cellodextrins. These may prove valuable targets in the genetic engineering of P. echinulatum focused on industrial cellulases production. Our study marks an important step in characterizing and understanding the molecular mechanisms employed by P. echinulatum in the enzymatic hydrolysis of lignocellulosic biomass.

Effect of alkali-treated birch sawdust on the lignocellulase secretion and exo-polysaccharide production by Inonotus obliquus under submerged fermentation and its lignocellulose degradation patterns.

The objectives of this study were to investigate the medicinal mushroom Inonotus obliquus on the production of polysaccharides and changes of extracellular lignocellulolytic enzymes during submerged fermentation using alkali-treated birch sawdust as substrate. Meanwhile, in order to explore the degradation mode of lignocellulose in alkali-treated birch sawdust, degradation analysis of three components of lignocellulose was carried out. The fungus process in alkali-treated birch sawdust medium resulted in a higher degradation rate of cellulose, hemicellulose, and lignin of 39.24%, 51.00% and 31.3% after 11 days of submerged fermentation by the mycelium of I. obliquus, respectively. Maximal polysaccharide production and α-glucosidase inhibition rate determined in the alkali-treated birch sawdust medium were 6.93 mg/mL and 55.80%, while they were 4.98 mg/mL and 27.89% in the control. Moreover, high activities of laccase (51.95 IU/mL), CMCase (1.35 IU/mL), filter paper activity (0.50 IU/mL) and ß-glucosidase (0.55 IU/mL) were observed in alkali-treated birch sawdust medium, respectively. The results demonstrated that the addition of alkali-treated birch sawdust could promote the yield and α-glucosidase inhibition activity of polysaccharides and induce the production of cellulase and xylanase, indicating that alkali pretreatment was conducive to utilization of birch sawdust by I. obliquus.

Degradation of lignocellulosic matrix of oilseed cakes by solid-state fermentation: fungi screening for enzymes production and antioxidants release.

BACKGROUND: Vegetable oils are yearly produced in large amounts generating solid by-products, the oilseed cake (OC). OCs are lignocellulosic materials that have been used for animal feed with some limitations due to high fibre content from the plant cell walls. Biotechnological processes can help to overcome these limitations and contribute to up-grading such by-products, enhancing their nutritional value as feed ingredients. RESULTS: All fungal species were able to decrease neutral detergent fibre and acid detergent fibre in all by-products. Additionally, relevant enzymes were produced by the three fungi studied resulting in an improved antioxidant capacity of all fermented OCs. Aspergillus niger led to the highest activity of cellulase (109 U g-1 ), xylanase (692 U g-1 ) and protease (157 U g-1 ) per dry OC matter and to the recovery of an extract rich in antioxidants, with the highest scavenging potential of free radicals and superoxide anion, iron chelation ability and reducing power. Rhyzopus oryzae produced the highest activity of ß-glucosidase (503 U g-1 ) and led to the highest liberation of total phenolic content (TPC). Principal components analysis showed that extracts with high antioxidant potential were obtained in solid-state fermentation (SSF) with high enzymatic activity. A positive correlation was established between the action of ß-glucosidase and TPC. CONCLUSION: Within the same bioprocess it was possible to improve the nutritional value of OCs and to obtain relevant bioactive compounds such as lignocellulosic enzymes and phenolic compounds with antioxidant potential, resulting in a significant improvement of already valuable by-products with commercial interest for animal feed. © 2021 Society of Chemical Industry.

Saccharification of lignocellulosic biomass using an enzymatic cocktail of fungal origin and successive production of butanol by Clostridium acetobutylicum.

A multistep approach was undertaken for biobutanol production targeting valorization of agricultural waste. Optimum production of lignocellulolytic enzymes [CMCase (3822.93U/mg), FPase (3640.93U/mg), ß-glucosidase (3873.92U/mg), xylanase (3460.24U/mg), pectinase (3359.57U/mg), α-amylase (4136.54U/mg), and laccase (3863.16U/mg)] was accomplished through solid-substrate fermentation of pretreated mixed substrates (wheat bran, sugarcane bagasse and orange peel) by Aspergillus niger SKN1 and Trametes hirsuta SKH1. Partially purified enzyme cocktail was employed for saccharification of the said substrate mixture into fermentable sugar (69.23 g/L, product yield of 24% w/w). The recovered sugar with vegetable extract supplements was found as robust fermentable medium that supported 16.51 g/L biobutanol production by Clostridium acetobutylicum ATCC824. The sequential bioprocessing of low-priced substrates and exploitation of vegetable extract as growth factor for microbial butanol production will open a new vista in biofuel research.

Overview of Lignocellulolytic Enzyme Systems with Special Reference to Valorization of Lignocellulosic Biomass.

Lignocellulosic biomass, one of the most valuable natural resources, is abundantly present on earth. Being a renewable feedstock, it harbors a great potential to be exploited as a raw material, to produce various value-added products. Lignocellulolytic microorganisms hold a unique position regarding the valorization of lignocellulosic biomass as they contain efficient enzyme systems capable of degrading this biomass. The ubiquitous nature of these microorganisms and their survival under extreme conditions have enabled their use as an effective producer of lignocellulolytic enzymes with improved biochemical features crucial to industrial bioconversion processes. These enzymes can prove to be an exquisite tool when it comes to the eco-friendly manufacturing of value-added products using waste material. This review focuses on highlighting the significance of lignocellulosic biomass, microbial sources of lignocellulolytic enzymes and their use in the formation of useful products.