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Microbial cellulases are highly versatile catalysts with significant potential in various industries, including pulp and paper, textile manufacturing, laundry, biofuel production, food and animal feed, brewing, and agriculture. Cellulases have attracted considerable attention from the scientific community owing to their broad industrial applications and the complex nature of enzymatic systems. In the present study, a novel fungal isolate of Aspergillus sp. IN5 was used to produce cellulases. We optimized each parameter, including carbon source, incubation temperature, pH, and incubation time, for maximum cellulase production using isolate IN5 under solid-state fermentation conditions. The optimized parameters for cellulase production by isolate IN5 under solid-state fermentation were as follows: substrate, soybean residue; incubation temperature, 35 °C; pH, 7.0; and incubation duration, 5 days. These conditions resulted in the highest total cellulase activity (0.26 U/g substrate), and carboxymethyl cellulase and ß-glucosidase activities of 3.32 and 196.09 U/g substrate, respectively. The obtained fungal cellulase was used for the enzymatic hydrolysis of acid- or alkali-pretreated rice straw, which served as a model substrate. Notably, compared with acid pretreatment, the pretreatment of rice straw with diluted alkali led to higher yields of reducing sugars. Maximum reducing sugar yield (286.06 ± 2.77 mg/g substrate) was obtained after 24-h incubation of diluted alkali-pretreated rice straw mixed with an enzyme loading of 15 U/g substrate. The findings of this study provide an alternative strategy for utilizing agricultural waste and an approach to efficiently produce cellulase for the degradation of lignocellulosic materials, with promising benefits for sustainable waste management.

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BACKGROUND: Marine cyanobacteria offer many sustainability advantages, such as the ability to fix atmospheric CO2, very fast growth and no dependence on freshwater for culture. Cyanobacterial biomass is a rich source of sugars and proteins, two essential nutrients for culturing any heterotroph. However, no previous study has evaluated their application as a feedstock for fungal bioprocesses. RESULTS: In this work, we cultured the marine cyanobacterium Synechococcus sp. PCC 7002 in a 3-L externally illuminated bioreactor with working volume of 2 L with a biomass productivity of ~ 0.8 g L-1 day-1. Hydrolysis of the biomass with acids released proteins and hydrolyzed glycogen while hydrolysis of the biomass with base released only proteins but did not hydrolyze glycogen. Among the different acids tested, treatment with HNO3 led to the highest release of proteins and glucose. Cyanobacterial biomass hydrolysate (CBH) prepared in HNO3 was used as a medium to produce cellulase enzyme by the Penicillium funiculosum OAO3 strain while CBH prepared in HCl and treated with charcoal was used as a medium for citric acid by Aspergillus tubingensis. Approximately 50% higher titers of both products were obtained compared to traditional media. CONCLUSIONS: These results show that the hydrolysate of marine cyanobacteria is an effective source of nutrients/proteins for fungal bioprocesses.

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BACKGROUND: The demand for low-cost cellulolytic enzyme synthesis is rising in the enzyme market. This work aims to produce cellulase by utilizing various agricultural wastes and investigating the use of enzyme in saccharification and textile industries. RESULTS: Solid state fermentation (SSF) was applied to produce industrial enzymes, particularly cellulase, through utilizing Molokhia (Corchorus olitorius) stems by Aspergillus awamori MK788209 isolate. Two stages of statistical factorial designs Plackett-Burman (PB) and Central Composite Design (CCD) were applied to enhance the A. awamori MK788209 cellulase production from Molokhia stems (MS). The fold increase of enzyme production by PB followed by CCD was 2.51 and 4.86, respectively. Additionally, the A. awamori MK788209 culture filtrate was highly effective in saccharifying various agricultural wastes, particularly pea peels (PP) (yielding 98.33 mg reducing sugar/ml), due to its richness in cellulase, laccase, xylanase, pectinase, and amylase. By optimizing the three main variables; pea peel weight, culture filtrate volume added, and saccharification time by CCD, the sugar recovery from PP was enhanced, leading to a 3.44-fold increase in reducing sugar recovery (338 mg reducing sugar /ml). Furthermore, the A. awamori MK788209 culture filtrate showed high efficacy in textile applications, enhancing the roughness, weight loss, white index, and printing capability of treated cotton fabrics. CONCLUSIONS: A. Awamori MK788209 produced cellulase which was effective in PP saccharification. The enzyme was also capable of enhancing cotton fabric properties.

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Cellulase is a new research point besides glucoamylase, amylase, and protease in the enzyme industry. Cellulase can decompose lignocellulosic biomass into small-molecule sugars, which facilitates microbial utilization; thus, it has a vast market potential in the field of feed, food, energy, and chemistry. The Aspergillus was the first strain used in cellulase preparation because of its safety and non-toxicity, strong growth ability, and high enzyme yield. This review provides the latest research and advances on preparing cellulase from Aspergillus. The metabolic mechanisms of cellulase secretion by Aspergillus, the selection of fermentation substrates, the comparison of the fermentation modes, and the effect of fermentation conditions have been discussed in this review. Also, the subsequent separation and purification techniques of Aspergillus cellulase, including salting out, organic solvent precipitation, ultrafiltration, and chromatography, have been declared. Further, bottlenecks in Aspergillus cellulase preparation and corresponding feasible approaches, such as genetic engineering, mixed culture, and cellulase immobilization, have also been proposed in this review. This paper provides theoretical support for the efficient production and application of Aspergillus cellulase.

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Development of low-cost and economic cellulase production is among the key challenges due to its broad industrial applications. One of the main topics of research pertaining to sustainable biomass waste based biorefinaries is the development of economic cellulase production strategies. The main cause of the increase in cellulase production costs is the use of commercial substrates; as a result, the cost of any cellulase-based bioprocess can be decreased by employing a productive, low-cost substrate. The goal of the current study is to develop low-cost cellulase using the carbohydrate-rich, renewable, and widely accessible cyanobacteria algae Oscillatoria obscura as the production substrate. Maximum cellulase was produced utilising the fungus Rhizopus oryzae at substrate concentration of 7.0 g among various tested concentrations of algal biomass. Maximum production rates of 22 IU/gds FP, 105 IU/gds BGL, and 116 IU/gds EG in 72 h were possible under optimal conditions and substrate concentration. Further investigations on the crude enzyme's stability in the presence of iron oxide nanoparticles (IONPs) revealed that it was thermally stable at 60 °C for up to 8 h. Additionally, the crude enzyme demonstrated pH stability by maintaining its complete activity at pH 6.0 for 8 h in the presence of the optimal dose of 15 mg IONPs. The outcomes of this research may be used to investigate the possibility of producing such enzymes in large quantities at low cost for industrial use.

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The present review explores fruit wastes as potential and low-cost substrates for economical production of cellulase enzymes. Being renewable, vast availability and having rich organic nutrient, these fruit wastes can be exploited to produce cellulase enzyme for various industrial applications. This review aimed to explore recent insight in sustainable production of microbial cellulolytic enzymes following solid state fermentation (SSF) wherein different types of fruit wastes as a potentially viable and alternative form of substrates have been utilized. In addition, detailed about the characteristics, mechanisms and market scenario of cellulase enzymes produced through a range of microbial species have been discussed. Further, impacts of different physicochemical parameters on solid-state fermentation based enzyme production and scale up issues have also analyzed. Moreover, applications of cellulases to produce different types of biofuels have been evaluated while emphases are made on existing hindrances and the possible strategies to improve the enzyme production process using fruit wastes.

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Fruit wastes can be imperative to elevate economical biomass to biofuels production process at pilot scale. Because of the renewable features, huge availability, having low lignin content organic nature and low cost; these wastes can be of much interest for cellulase enzyme production. This review provides recent advances on the fungal cellulase production using fruit wastes as a potential substrate. Also, the availability of fruit wastes, generation and processing data and their potential applications for cellulase enzyme production have been discussed. Several aspects, including cellulase and its function, solid-state fermentation, process parameters, microbial source, and the application of enzyme in biofuels industries have also been discussed. Further, emphasis has been made on various bottlenecks and feasible approaches such as use of nanomaterials, co-culture, molecular techniques, genetic engineering, and cost economy analysis to develop a low-cost based comprehensive technology for viable production of cellulase and its application in biofuels production technology.

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BACKGROUND AND OBJECTIVE: Cellulase as a fibrolytic enzyme is a highly effective tool for agricultural waste treatments. Production of cellulase enzyme on medium of agricultural wastes by Fusarium graminearum to be used in ruminant feeding was the main objective of this study. MATERIALS AND METHODS: Impact of initial pH of growth medium, different nitrogen sources and variety of agriculture by products as a carbon sources on cellulase production have been studied. Electron microscope was used for investigate the impact of the resultant cellulase on corn stover degradation, while batch culture technique was used for investigate impact of different levels of the produced and commercial cellulases on total mixed ration digestibility by rumen microorganisms (in vitro). RESULTS: Cellulase maximum production by F. graminearum was obtained at 20% corn stover, initial pH of growth medium 5.0 and peptone as a nitrogen source. All addition levels of the produced cellulase increased dry matter (DM), neutral detergent fiber (NDF), acid detergent fiber (ADF), cellulose and hemicellulose degradability of the treated diets, but the maximum produced cellulase efficiency% for dry matter degradability was obtained at 1200 IU kg-1 DM reached 23.19% over the control. CONCLUSION: Utilization of the produced cellulase in enrichment of the feeding value of the agricultural by-products may help in overcome of the feed gap with good impact on environment and public health.

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The aim of this study was to investigate the effect of pH control by CO 2 pressurization on the enzymatic hydrolysis of herbaceous feedstock in the calcium capturing by carbonation (CaCCO) process for fermentable sugar production. The pH of the slurry of 5 % (w/w) Ca(OH) 2 -pretreated/CO 2 -neutralized rice straw could be controlled between 5.70 and 6.38 at 50 °C by changing the CO 2 partial pressure ( p CO 2 ) from 0.1 to 1.0 MPa. A mixture of fungal enzyme preparations, namely, Trichoderma reesei cellulases/hemicellulases and Aspergillus niger ß-glucosidase, indicated that pH 5.5-6.0 is optimal for solubilizing sugars from Ca(OH) 2 -pretreated rice straw. Enzymatic saccharification of pretreated rice straw under various p CO 2 conditions revealed that the highest soluble sugar yields were obtained at p CO 2 0.4 MPa and over, which is consistent with the expected pH at the p CO 2 without enzymes and demonstrates the effectiveness of pH control by CO 2 pressurization.

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Background: The recalcitrance of cellulosic biomass is widely recognized as a key barrier to cost-effective biological processing to fuels and chemicals, but the relative impacts of physical, chemical and genetic interventions to improve biomass processing singly and in combination have yet to be evaluated systematically. Solubilization of plant cell walls can be enhanced by non-biological augmentation including physical cotreatment and thermochemical pretreatment, the choice of biocatalyst, the choice of plant feedstock, genetic engineering of plants, and choosing feedstocks that are less recalcitrant natural variants. A two-tiered combinatoric investigation of lignocellulosic biomass deconstruction was undertaken with three biocatalysts (Clostridium thermocellum, Caldicellulosiruptor bescii, Novozymes Cellic® Ctec2 and Htec2), three transgenic switchgrass plant lines (COMT, MYB4, GAUT4) and their respective nontransgenic controls, two Populus natural variants, and augmentation of biological attack using either mechanical cotreatment or cosolvent-enhanced lignocellulosic fractionation (CELF) pretreatment. Results: In the absence of augmentation and under the conditions tested, increased total carbohydrate solubilization (TCS) was observed for 8 of the 9 combinations of switchgrass modifications and biocatalysts tested, and statistically significant for five of the combinations. Our results indicate that recalcitrance is not a trait determined by the feedstock only, but instead is coequally determined by the choice of biocatalyst. TCS with C. thermocellum was significantly higher than with the other two biocatalysts. Both CELF pretreatment and cotreatment via continuous ball milling enabled TCS in excess of 90%. Conclusion: Based on our results as well as literature studies, it appears that some form of non-biological augmentation will likely be necessary for the foreseeable future to achieve high TCS for most cellulosic feedstocks. However, our results show that this need not necessarily involve thermochemical processing, and need not necessarily occur prior to biological conversion. Under the conditions tested, the relative magnitude of TCS increase was augmentation > biocatalyst choice > plant choice > plant modification > plant natural variants. In the presence of augmentation, plant modification, plant natural variation, and plant choice exhibited a small, statistically non-significant impact on TCS.

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Bioprocessing of lignocellulose as a renewable resource for fuels, chemicals or value added products is a necessity to fulfil demands of petroleum products. This study aims to convert corn stover to polyhydroxyalkanoates (PHA). Corn stover was hydrolyzed to crude sugars by an on-site prepared cellulase cocktail from co-culture of Trichoderma reesei and Aspergillus niger. The potent PHA producer, Paracoccus sp. LL1, was isolated from Lonar Lake, India and could accumulate PHA up to 72.4% of its dry cell weight. PHA production reached 9.71 g/L from corn stover hydrolysate containing 40 g/L sugar mixture. The PHA synthase gene (phaC) sequence of the isolate showed 79% identity with the phaC gene of Paracoccus seriniphilus (E71) strain from the NCBI database. The nature/type of PHA was found to be poly(3-hydroxybutyrate) by Fourier transform infrared spectroscopy.

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Plant-pathogenic fungi produce cellulases. However, little information is available on cellulase as an elicitor in plant-pathogen interactions. Here, an endocellulase (EG1) was isolated from Rhizoctonia solani. It contains a putative protein of 227 amino acids with a signal peptide and a family-45 glycosyl hydrolase domain. Its aspartic acid (Asp) residue at position 32 was changed to alanine (Ala), resulting in full loss of its catalytic activity. Wild-type and mutated forms of the endoglucanase were expressed in yeast and purified to homogeneity. The purified wild-type and mutant forms induced cell death in maize, tobacco and Arabidopsis leaves, and the transcription of three defence marker genes in maize and tobacco and 10 genes related to defence responses in maize. Moreover, they also induced the accumulation of reactive oxygen species (ROS), medium alkalinization, Ca(2+) accumulation and ethylene biosynthesis of suspension-cultured tobacco cells. Similarly, production of the EG1 wild-type and mutated forms in tobacco induced cell death using the Potato virus X (PVX) expression system. In vivo, expression of EG1 was also related to cell death during infection of maize by R. solani. These results provide direct evidence that the endoglucanase is an elicitor, but its enzymatic activity is not required for its elicitor activity.

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