Endoglucanase Production Research Articles

Mass production and characterization of an endoglucanase from Coleoptera insect (Monochamus saltuarius) in yeast Kluyveromyceslactis

To harness the diverse industrial applications of cellulase, including its use in the food, pulp, textile, agriculture, and biofuel sectors, this study focused on the high-yield production of a bioactive insect-derived endoglucanase, Monochamus saltuarius glycoside hydrolase family 5 (MsGHF5). MsGHF5 was introduced into the genome of Kluyveromyces lactis to maintain expression stability, and mass production of the enzyme was induced using fed-batch fermentation. After 40 h of cultivation, recombinant MsGHF5 was successfully produced in the culture broth, with a yield of 29,000 U/L, upon galactose induction. The optimal conditions for the activity of purified MsGHF5 were determined to be a pH of 5 and a temperature of 35 °C, with the presence of ferrous ions enhancing the enzymatic activity by up to 1.5-fold. Notably, the activity of MsGHF5 produced in K. lactis was significantly higher than that produced in Escherichia coli, suggesting that glycosylation is crucial for the functional performance of the enzyme. This study highlights the potential use of K. lactis as a host for the production of bioactive MsGHF5, thus paving the way for its application in various industrial sectors.

Production and partial characterization of endoglucanase by Thermothelomyces heterothallicus PA2S4T and its application in biopolishing of denim jeans

Production and partial characterization of endoglucanase by Thermothelomyces heterothallicus PA2S4T and its application in biopolishing of denim jeans

Optimization of hydrolases production by Penicillium crustosum in submerged fermentation using agro-waste residues as cosubstrate

This study aimed to enhance the production of exo-polygalacturonases (exo-PG), xylanases, and endoglucanases using Penicillium crustosum through submerged fermentation. Lemon peel (LP) and wheat bran (WB) were examined as potential cosubstrate for hydrolase production. The highest exo-PG and endoglucanase activities were observed with LP, whereas xylanase activity showed improvement with the addition of WB. To optimize exo-PG and endoglucanase production, a 23 Face-Centered Central Composite Design (FCCCD) was employed, varying crucial physicochemical factors. Similarly, a 24 FCCCD was utilized to optimize xylanase production by varying temperature, pH, MnSO4 concentration as a cofactor, and the PDB culture medium. Under optimized conditions, exo-PG production reached 1427.99 ± 27.41 U/mL, marking a 45% increase compared to initial fermentation conditions. Endoglucanase production peaked at 62.68 ± 9.65 U/mL under conditions of 45 °C, pH 5.5, and 2.5 g/L MnSO4. Xylanase production reached 271.77 ± 8.50 U/mL under optimal conditions (37.5 °C, pH 5.5, 2.5 g/L MnSO4, and 62.5% PDB). The optimized simultaneous production of the hydrolytic cocktail occurred at 45 °C, pH 5.5, and 2.5 g/L MnSO4 with LP, yielding maximum activities of 959.74 U/mL for exo-PG, 61.98 U/mL for endoglucanases, and 65.60 U/mL for xylanases as determined by FCCCD. In conclusion, Penicillium crustosum shows promise as a hydrolase producer, and the utilization of agro-wastes as cosubstrate enhances the yield of enzymes with industrial relevance.

Novel strategy to elevate solid state fermentation to produce alkilophilic endoglucanase using date waste feedstocks and peapod extract based nutrient media and expired probiotic strain: Application in fermentable sugar production

Solid waste generation and its impactful management are among the most raised environmental concerns seeking sustainable alternatives in the form of value addition. In this study, a novel strategy has been presented towards enhanced bacterial endoglucanase (EG) production under solid state fermentation (SSF). The EG was produced using the waste probiotic juice of the bacteria Lactobacillus casei strain Shirota on a substrate of waste date pulp and seeds in co-fermentation mode. Additionally, peapod extract has been applied as a nutrient medium to maintain moisture in place of commercial media in SSF. Further, for enhancement in enzyme production, the impact of incubation temperature and pH of the SSF medium have been studied to obtain optimum enzyme activity. Under optimum conditions, bacteria produced 17.2 U/mL EG at a temperature of 37 °C and a pH of 8.0. The raw EG showed pH and thermal stability up to 5 h at 60 °C and pH 8.0. Furthermore, the enzymatic hydrolysis of date pulp released 67.71 g/L fermentable glucose obtained in 24 h of the reaction at 60 °C and pH 8.0. This work may have great significance in the areas of solid waste management and microbe-mediated value addition of waste biomass in the context of hydrolytic enzyme production for biorefinery applications.

Conversion of Corn Stover for Microbial Enzymes Production by Phanerochaete chrysosporium.

Phanerochaete chrysosporium, a white rot fungus, exhibits remarkable capabilities in producing various extracellular enzymes. These microbial enzymes find extensive applications in disrupting the intricate structure of plant cell walls, decolorizing synthetic dyes, and facilitating pulp extraction, among other functions. The process of solid-state fermentation stands out as an economical and sustainable approach, ideal for achieving high yields in enzyme production using lignocellulosic biomass as a substrate. In this research paper, both untreated and alkali pretreated corn stover materials served as substrates for enzyme production, leveraging the fungal strain's capacity to generate enzymes like cellulases and manganese peroxidase. The maximum production of endoglucanase was notably observed, reaching 121.21 ± 0.90 U/gds on the 9th day for untreated biomass and 79.75 ± 0.57 U/gds on the 6th day for treated biomass. Similarly, the peak exoglucanase production was recorded at 2.46 ± 0.008 FPU/ml on the 3rd day for untreated biomass and 0.92 ± 0.002 FPU/ml on the 6th day for treated biomass. Furthermore, the highest production of manganese peroxidase was achieved, with values of 5076.81 U/l on the 6th day for untreated biomass and 1127.58 ± 0.23 U/l on the 3rd day for treated biomass. These results collectively emphasize the potential of corn stover as a renewable and promising substrate for the production of essential enzymes.

Pathway engineering of Escherichia coli for the glycerol-based production of recombinant endoglucanase

BackgroundTechnology platforms developed in biorefinery are featured with low-carbon emission but afflicted by the issue of economic viability. This can be addressed by a cascade process that prioritizes the production of value-added products. In this context, the cost-effective production of endoglucanase (encoded by celA) was illustrated using Escherichia coli with crude glycerol. This renewable and sustainable feedstock appears in abundance in waste streams of the biodiesel process. Moreover, glutamate (Glu) was supplemented to tackle the inconsistent quality of yeast extract. MethodsA new fusion tag was first explored to improve the solubility of CelA. CelA was then produced using a strain quipped with a rewired glycerol metabolism. To improve the CelA production, the producer strain was further re-programmed by manipulation of metabolic pathways involved in oxidative phosphorylation and substrate-level phosphorylation. Significant findingsThe production of CelA in the developed strain positively correlated with glycerol. It was found to increase the CelA production by 90% in the engineered strain when the glycerol level was doubled. In contrast, the parent strain gained merely 20% more production of CelA. The result indicates that the physiological status of the engineered strain is refined to favor functioning of the protein synthesis machinery. In conclusion, the developed strain shows promise in production of recombinant proteins based on crude glycerol.

Utilization of spent substrates and waste products of mushroom cultivation to produce new crops of Pleurotus ostreatus, Pleurotus eryngii and Agaricus bisporus

Five agro-industrial residues, their corresponding spent mushroom substrates (SMS), commercial fresh and spent mushroom compost of Agaricus (SMC) and Pleurotus ostreatus (SMS GZ) cultivation, Pleurotus and Agaricus waste (stipes/mishappen mushrooms) were combined and re-utilized as novel substrates for the cultivation of P. ostreatus, P. eryngii and A. bisporus mushrooms. SMSs/ SMC were used up to 40% (treatment 1 and 2), while 20% of mushroom waste were added in fresh and spent substrates (treatment 3 and 4). The impact of different substrates on mycelial growth rate and biomass production was examined. Then, Pleurotus spp. were cultivated on the most promising substrates and essential cultivation aspects (earliness, total mushroom yield, biological efficiency-BE) and carposomes’ quality parameters (weight, morphological characteristics) were evaluated. Laccase and endoglucanase production by Pleurotus species were also determined at 50 and 100% of colonization stages. All species showed their fastest mycelial growth rate (up to 5 mm/day) on substrates consisted of SMC, whereas many combinations of species/substrate enhanced biomass production. The SMS GZ supplementation positively affected laccase activity; in the cultures of P. ostreatus and P. eryngii the highest values were 62,539 and 17,584 U/g d.w., respectively. On the contrary, small amounts of endoglucanase were produced (0.007 to 0.322 U/g d.w.); the greatest production was recorded for P. ostreatus at full colonization. Regarding fermentation in bags, significant amount of total mushroom yield was produced in all substrates and those with SMS GZ supported the fastest earliness period and the highest BE for both Pleurotus species. BE values ranged from 54 to 133% for P. ostreatus and from 53 to 121% for P. eryngii. Concerning morphological characteristics, mushroom waste addition seemed to affect them positively. The data included in this paper support the effective re-utilization of different types of SMS and mushroom waste for fungal mass and enzymes’ production and for new high quality Pleurotus spp. carposomes.

Production, Purification, Characterization, and Application of Halotolerant and Thermostable Endoglucanase Isolated from Pycnoporus sanguineus

Production, Purification, Characterization, and Application of Halotolerant and Thermostable Endoglucanase Isolated from Pycnoporus sanguineus

Production of the Cellulase Enzyme System by Locally Isolated Trichoderma and Aspergillus Species Cultivated on Banana Pseudostem during Solid-State Fermentation

The production cost of cellulases is regarded as a limiting factor in the cellulosic ethanol production chain. Trichoderma and Aspergillus species were used to produce cellulases through solid-state fermentation (SSF) utilizing banana pseudostem (BPS) as a carbon source. The production of cellulases was investigated at various substrate moisture contents (65–80%), incubation temperatures (30–40 °C), substrate pre-treatment methods (3% w/v NaOH, 5% v/v H2SO4, and water), and with different co-culturing of microorganisms. Trichoderma longibrachiatum LMLSAUL 14-1 produced the maximum total cellulase (75 FPU/g d.s), endoglucanase (11.35 U/g d.s), and β-glucosidase (235.83 U/g d.s) activities at a 75% moisture content of the untreated BPS at 30 °C in static culture. Pre-treatment of BPS improved the production of specific enzymes. Aspergillus fumigatus LMLPS 13-4 produced more β-glucosidase (259.8 U/g d.s) when grown on acid-pre-treated BPS, whereas T. harzianum LMLBP07 13-5 produced the highest β-glucosidase activity (319.5 U/g d.s) on alkali-pre-treated BPS. Co-culturing of T. harzianum LMLBP07 13-5 and A. fumigatus LMLPS 13-4 improved the production of endoglucanase. These results suggest that banana pseudostem, a waste product of the banana industry, could be a potentially cheaper and abundant substrate for the production of the cellulase enzymes.

Bacterial cellulase production via co-fermentation of paddy straw and Litchi waste and its stability assessment in the presence of Zn[sbnd]Mg mixed-phase hydroxide-based nanocomposite derived from Litchi chinensis seeds

Co-fermentation via co-cultured bacterial microorganisms to develop enzymes in solid-state fermentation (SSF) is a promising approach. This strategy is imperative in a series of sustainable and effective approaches due to superior microbial growth and the use of a combination of inexpensive feedstocks for enzyme production wherein mutually participating enzyme-producing microbial communities are employed. Moreover, the addition of nanomaterials to this technique may aid in its prominent advantage of enhancing enzyme production. This strategy may be able to decrease the overall cost of the bioprocessing to produce enzymes by further implementing biogenic, route-derived nanomaterials as catalysts. Therefore, the present study attempts to explore endoglucanase (EG) production using a bacterial coculture system by employing two different bacterial strains, namely, Bacillus subtilis and Serratia marcescens under SSF in the presence of a ZnMg hydroxide-based nanocomposite as a nanocatalyst. The nanocatalyst based on ZnMg hydroxide has been prepared via green synthesis using Litchi waste seed, while SSF for EG production has been conducted using cofermentation of litchi seed (Ls) and paddy straw (Ps) waste. Under an optimized substrate concentration ratio of 5:6 Ps:Ls and in the presence of 2.0 mg of nanocatalyst, the cocultured bacterial system produced 1.6 IU/mL of EG enzyme, which was ~1.33 fold higher as compared to the control. Additionally, the same enzyme showed its stability for 135 min in the presence of 1.0 mg of nanocatalyst at 38 °C. The nanocatalyst has been synthesized using the green method, wherein waste litchi seed is used as a reducing agent, and the nanocatalyst could be employed to improve the production and functional stability of crude enzymes. The findings of the present study may have significant application in lignocellulosic-based biorefinaries and cellulosic waste management.

CRISPR-Based Multi-Gene Integration Strategies to Create Saccharomyces cerevisiae Strains for Consolidated Bioprocessing

Significant engineering of Saccharomyces cerevisiae is required to enable consolidated bioprocessing (CBP) of lignocellulose to ethanol. Genome modification in S. cerevisiae has been successful partly due to its efficient homology-directed DNA repair machinery, and CRISPR technology has made multi-gene editing even more accessible. Here, we tested the integration of cellulase encoding genes to various sites on the yeast genome to inform the best strategy for creating cellulolytic strains for CBP. We targeted endoglucanase (EG) or cellobiohydrolase (CBH) encoding genes to discreet chromosomal sites for single-copy integration or to the repeated delta sites for multi-copy integration. CBH1 activity was significantly higher when the gene was targeted to the delta sequences compared to single gene integration loci. EG production was comparable, though lower when the gene was targeted to a chromosome 10 site. We subsequently used the information to construct a strain containing three cellulase encoding genes. While individual cellulase activities could be assayed and cellulose conversion demonstrated, it was shown that targeting specific genes to specific loci had dramatic effects on strain efficiency. Since marker-containing plasmids could be cured from these strains, additional genetic changes can subsequently be made to optimize strains for CBP conversion of lignocellulose.

Nghiên cứu biểu hiện endo-glucanase tái tổ hợp nguồn gốc từ DNA metagenom vi khuẩn dạ cỏ dê ở quy mô bình lên men 5 lít

Cellulose is one of the three main components of lignocellulose and one of the world's largest reserves of biomass. Endo-glucanase is one of the three crucial enzyms in the hydrolysis process of cellulose into glucoses. The endo-glucanase gene has been mined by bioinformatics tools in next-generation sequencing data of metagenom DNA from bacteria residing in goat rumen. In previous study, the endo-glucanase gene was successfully recombinantly expressed in the E. coli Rosetta1 strain in flask, the expressed protein existed in the soluble form with good activity. In order to apply in bigger scale, endo-glucanase was investigated for expression in a 5-liter bio-reactor. The research results showed that the expressed protein reached about 1.5-1.7 mg/ml with crude activity of about 37-38 U/ml of fermentation solution. The endo-glucanase protein, after being purified by His-tag affinity chromatography column and salt-removed by membrane dialysis, has a fairly high activity of about 400 U/mg. This result showed potential for the application of endo-glucanase production on an industrial scale.

Multi-efficient endoglucanase from Aspergillus niger MPS25 and its potential applications in saccharification of wheat straw and waste paper deinking

The deinking in the paper industry is in great demand, and microbial enzymes are key players. In the present study, the endoglucanase production from newly isolated fungi Aspergillus niger MPS25 is reported. The optimization of endoglucanase production was carried out using one factor at a time approach resulting in endoglucanase activity (2.37 IU/ml) at 45 °C and pH 8 in submerged fermentation (SmF), which shows optimum enzyme activity at 60 °C. Interestingly, the metal ions viz. Co2+ stimulated the endoglucanase activity, whereas Mn2+ reduced the enzyme activity, which shows that this enzyme can be used for effluent treatment released through deinking. The enzymatic hydrolysis of wheat straw produced 26.96 ± 0.108 mg/g of reducing sugars, indicating its potential in saccharification and the biofuel industry. Furthermore, the validation of the deinking efficiency of this enzyme resulted in improved deinking of mixed office waste and old newspapers by 31.5% and 20.4%, respectively. The strength properties, viz. burst factor and tear index, breaking length, and tensile index of the handmade paper sheets, were also improved which were analyzed by the scanning electron micrographs. The FTIR and XRD analysis of pulp provided insights into the changes in functional groups and cellulose crystallinity, respectively. These results indicate that multi-efficient endoglucanase from Aspergillus niger MPS25 is suitable for enzyme-based eco-friendly deinking for waste paper recycling and lignocellulosic biomass saccharification.

Immobilization of Recombinant Endoglucanase (CelA) from Clostridium thermocellum on Modified Regenerated Cellulose Membrane

Cellulases are being widely employed in lignocellulosic biorefineries for the sustainable production of value-added bioproducts. However, the high production cost, sensitivity, and non-reusability of free cellulase enzymes impede their commercial applications. Enzyme immobilization seems to be a potential approach to address the aforesaid complications. The current study aims at the production of recombinant endoglucanase (CelA) originated from the cellulosome of Clostridium thermocellum in Escherichia coli (E. coli), followed by immobilization using modified regenerated cellulose (RC) membranes. The surface modification of RC membranes was performed in two different ways: one to generate the immobilized metal ion affinity membranes RC-EPI-IDA-Co2+ (IMAMs) for coordination coupling and another to develop aldehyde functional group membranes RC-EPI-DA-GA (AMs) for covalent bonding. For the preparation of IMAMs, cobalt ions expressed the highest affinity effect compared to other metal ions. Both enzyme-immobilized membranes exhibited better thermal stability and maintained an improved relative activity at higher temperatures (50–90 °C). In the storage analysis, 80% relative activity was retained after 15 days at 4 °C. Furthermore, the IMAM- and AM-immobilized CelA retained 63% and 53% relative activity, respectively, after being reused five times. As to the purification effect during immobilization, IMAMs showed a better purification fold of 3.19 than AMs. The IMAMs also displayed better kinetic parameters, with a higher Vmax of 15.57 U mg−1 and a lower Km of 36.14 mg mL−1, than those of AMs. The IMAMs were regenerated via treatment with stripping buffer and reloaded with enzymes and displayed almost 100% activity, the same as free enzymes, up to 5 cycles of regeneration.

Pretreatment of lignocellulosic biogas substrates by filamentous fungi

Decomposition of lignocellulosic plant biomass by four filamentous fungi was carried out to facilitate subsequent anaerobic degradation and biogas formation. Agricultural side products, wheat straw and corn stover and forestry energy plant willow chips were selected as plant biomass sources. The substrates were confronted by pure cultures of Penicillium aurantiogriseum (new isolate from rumen), Trichoderma reesei (DSM768), Gilbertella persicaria (SZMC11086) and Rhizomucor miehei (SZMC11005). In addition to total cellulolytic filter paper degradation activity, the production of endoglucanase, cellobiohydrolase, β-glucosidase enzymes were followed during the pretreatment period, which lasted for 10 days at 37 °C. The products of pretreatments were subsequently tested for mesophilic biogas production in batch reactors. All 4 strains effectively pretreated the lignocellulosic substrates albeit in varying degrees, which was related to the level of the tested hydrolytic enzyme activities. Penicillium aurantiogriseum showed outstanding hydrolytic enzyme production and highest biogas yield from the partially degraded substrates. Corn stover was the best substrate for biomass decomposition and biogas production. Scanning electron microscopy confirmed the deep penetration of fungal hyphae into the lignocellulosic substrate in all cases.

Screening of quorum sensing and biofilm inhibitors among pseudomonas aeruginosa clinical isolates

Quorum sensing (QS) is a process that enables bacteria to communicate using secreted small pheromone-like signaling molecules called autoinducers (AIs). This process enables a population of bacteria to regulate gene expression collectively. These quorum sensing systems regulates biofilm formation, virulence factors expression, bioluminescence, motility patterns, exopolysaccharide production, antifungal or antibiotic production, endoglucanase production, pigmentation, competence, plasmid conjugal transfer and cross-signaling between strains and species. P. aeruginosa is an important and very dangerous opportunistic pathogen causing many fatal infections in patients with serious medical conditions. P. aeruginosa is associated with nosocomial infections mainly in immunocompromised patients, and it is rated the third-most-common microorganism associated with hospital-acquired infections representing about 10-15% of nosocomial infections recorded globally. Moreover, qRT-PCR revealed a significant reduction in expression of quorum sensing genes in virulence inhibitors-treated P. aeruginosa in comparison with untreated bacteria. Virulence inhibitors could play a role in reduction of Pseudomonas quorum sensing-dependent virulence factors production such as biofilm production, and therefore affect its pathogenesis in the host.

Alternative strategies to perform solid-state cultivation in a multilayer packed-bed bioreactor: Continuous and cyclic operations

The packed-bed bioreactor (PBB) is one of the equipment most used in solid-state cultivation (SSC) processes. However, PBBs present problems in the removal of metabolic heat generated by microorganisms, making difficult to scale up these equipment, that are usually operated in batches. Aiming to minimize the overheating problems and to provide more versatility for biorefineries, this work aimed to evaluate alternative modes of operation of a multilayer PBB applied in the production of endoglucanase by SSC, namely cyclic and continuous operations. For this purpose, the fungus Myceliophthora thermophila I-1D3b was used as inoculum and sugarcane bagasse and wheat bran were used as substrates. In the cyclic operation, the layers had their positions changed every 24 h. In the continuous process, newly inoculated layers were added and spent layers fermented were removed every 24 h, leading to a regular flow of layers. Maximum temperatures reached 48.1, 48.2 and 48.3 °C and endoglucanase productivities were 2.85, 3.18 and 3.35 U/g of dry solid/h for classical, cyclic and continuous operations, respectively. These results showed no significant difference according to the Tukey test (α = 0.05). Despite that, in the continuous operation, the maximum CO2 concentration recorded was 60% lower than in batch operations. In addition, it was possible to estimate the biomass production rate in continuous operation, which was 5.32 g/h. Therefore, the continuous operation proved to be applicable and advantageous, being able to contribute to the scale up and operational versatility of PBBs and, consequently, to the industrial advancement of SSC.

Degradative impact of Alternaria multiformis on novel polymeric biocomposites with the fillers of industrial waste materials under different pH and temperature conditions

The impact of Alternaria multiformis on novel polymeric biocomposites formed from epoxidized linseed oil and various types of fillers: pine needles (PN), pine bark (PB), grain mill waste (GW), rapeseed cake (RC) and specimen without filler (WF) under different pH and temperature conditions has been studied. Identification of fungal strain A. multiformis 0065 isolated from synthetic polymer was based on molecular and morphological analysis. Tested fungal strain showed significant production of polyphenol oxidase and weak production of amylase, endoglucanase, xylanase, and lipase. Polymeric biocomposites PB, GW, RC, and WF exposed to A. multiformis for four weeks showed the greatest weight loss at pH 4 or pH 5 and at a temperature of 26 °C, suggesting the optimal conditions for biodegradation of polymeric biocomposites studied. The attenuated total reflectance infrared spectroscopy (ATR-IR) and scanning electron microscopy (SEM) analysis confirmed biodegradation of polymeric biocomposites after fungal attack. ATR-IR technique revealed changes in the intensities of the signals, demonstrating dependence of degradation on the type of the filler. SEM analysis showed significant alterations of the specimen surfaces, indicating degradative impact of A. multiformis. Identification of the most biodegraded constituents could help in the creation of more environmentally friendly biocomposites.

Evaluation of thermostable endoglucanase in Paenibacillus lautus strain BHU3 for yield enhancement

In the present study, Paenibacillus lautus strain BHU3 isolated from landfill soil was evaluated for the presence of potential endoglucanases which are the first candidate of cellulase enzyme system to act on cellulose. In-silico analysis revealed high potential thermostable endoglucanases which can efficiently interact with cellulose. The most potent and thermostable endoglucanase (locus tag id. CPZ30_18280) belonged to glycosyl family-5 and had interaction energy of − 12.981 kcal/mol for the best docked cluster containing three out of ten docking conformations, and Tm value of 73.3 °C. MD simulation of 100 ns proved highly stable binding interactions of CPZ30_18280 endoglucanase with cellulose with root mean square deviation (RMSD) values ranging from 0.15 to 0.30 nm. Consistent interactions with characteristic active site residues (tyrosine, tryptophan and aspartate) of glycosyl family-5 endoglucanases were found. Further, to enhance the production of endoglucanases, the fermentation conditions were optimized employing approaches like one factor at a time (OFAT) and response surface methodology (RSM). Maximum activity of endoglucanase was determined at 60 °C. The optimized conditions for enhanced production of endoglucanase (10.15 U/mL) were pH 6.63, yeast extract conc. 3.44 g/L, wheat bran 3.59%, and inoculum size 2.65%. Hence, P. lautus strain BHU3 has enormous potential to synthesize highly efficient thermostable endoglucanases under optimized regime using agro-wastes. Thus, it could find immense industrial applications including large scale cellulose conversion to bioethanol.

Evaluation of endoglucanase and xylanase production by Aspergillus tamarii cultivated in agro-industrial lignocellulosic biomasses.

To better understand the production of enzymes of industrial interest from microorganisms with biotechnological potential using lignocellulosic biomass, we evaluated the production of endoglucanase and xylanase from Aspergillus tamarii. CAZymes domains were evaluated in the genome, and a screening of the enzymatic potential of A. tamarii in various agricultural biomasses was done. The enzymatic profile could be associated with the biomass complexity, with increased biomass recalcitrance yielding higher activity. A time-course profile defined 48h of cultivation as the best period for cultivating A. tamarii in sugarcane bagasse reached 12.05IU/mg for endoglucanase and 74.86IU/mg for xylanase. Using 0.1% (w/v) tryptone as the only nitrogen source and 12µmol/L CuSO4 addition had an overall positive effect on the enzymatic activity and protein production. A 22 factorial central composite design was used then to investigate the simultaneous influence of tryptone and CuSO4 on enzyme activity. Tryptone strongly affected enzymatic activity, decreasing endoglucanase activity but increasing xylanase activity. CuSO4 supplementation was advantageous for endoglucanases, increasing their activity, and it had a negative effect on xylanases. But overall, the experimental design increased the enzymatic activity of all biomasses used. For the clean cotton residue, the experimental design was able to reach the highest enzyme activity for endoglucanase and xylanase, with 1.195IU/mL and 6.353IU/mL, respectively. More experimental studies are required to investigate how the biomass induction effect impacts enzyme production.