Study on regulation of growth and biosynthesis of cellulolytic enzymes from newly isolated Aspergillus fumigatus ABK9 (original) (raw)
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International journal for research in applied sciences & Engineering Technology, 2019
Cellulase are a group of hydrolytic enzymes capable of hydrolysing the most abundant organic polymer i.e. cellulose to smaller sugar components including glucose subunits. In the current study the cellulolytic fungi were isolated from natural resources like cattle dung contaminated soil & cattle dung samples for cellulase production. The samples were collected from 3 different areas of valsad region. The primary screening of cellulolytic fungi were adopted by exposing Carboxyl Methyl Cellulose agar (CMC), which is a selective media for cellulolytic microorganisms, the 14 fungal isolates were screened. The Further screening was carried out on Czapek-Dox agar medium amended with 1% CMC; among 10 efficient Cellulolytic fungi were showing the Cellulase activity. Depending upon the maximum diameter of clear zone produced in CMC agar medium & Czapek-Dox agar medium 2 different fungi CPF-3 & CPF-8 were selected for higher production of one of the most highly demanded industrial enzyme cellulase using cheap and easily available agro wastes as sole source of carbon. The production of cellulase was substantially enhanced by optimizing the several parameters like pH, temperature, substrate concentration; nitrogen sources and agro based materials were evaluated under both fermentation conditions such as Solid State Fermentation (SSF) & Submerged Fermentation (SmF). The highest cellulase production was recorded in SSF medium than SmF by CPF-8 fungal isolate by optimizing the culture conditions. The optimum conditions found for cellulase production were achieved at 30℃ temperature, pH 5 with urea as nitrogen source and the agro waste material saw dust to stimulate the production of cellulase. After the fermentation process the enzymes were partially purified by ammonium sulphate precipitation method. The untreated substrate saw dust was hydrolysed with the partially purified enzyme cellulases and the hydrolysed product was fermented by the yeast for the bioethanol production. The cellulase activity was assayed by Carboxyl Methyl Cellulase "CMCase" assay & the protein estimation by Folin-Lowry method. The maximum cellulase activity of enzyme achieved by CPF-8 fungal culture 903.7 IU/ml in SSF & where as in SmF cellulase activity was achieved 800 IU/ml. The higher enzyme activity producer fungal isolate CPF-8 were further used for bioethanol production. After partial purification of enzyme the maximum enzyme activity was achieved in both fermentation condition at the 80% saturation about 192.59 IU/ml in SSF & 133.33 IU/ml was achieved in SmF medium. The untreated substrate saw dust hydrolysed by cellulase of SSF medium yielded at 120 hour of incubation period 2.05 mg ml-1 sugar which produced 1.4 g% ethanol on fermentation. The ethanol yield of the hydrolysate of substrate saw dust by cellulase of SmF medium lower about 1.2 g%. The without pre-treated substrate saw dust, hydrolysed with cellulase of SSF medium is established as suitable for bioethanol production.
International Journal of Plant & Soil Science
Cellulase enzymes are belonging to the hydrolytic group of enzymes facilitates the sugar release and its bioconversion into different valuable industrial products. Isolated micro fungi from rice straw by dilution plating pouring method studied for playing a various role in industries as well as in agriculture application. Various micro-fungi show enzymatic degradation of lignocellulosic material. The present study optimized the growing conditions for cellulase enzymes production from A.fumigatus. Optimization of various growth conditions such as temperature, different pH level and nitrogen source were studied for the production of enzyme carboxymethyl cellulase during this study. The result showed that A. fumigatus produced highest cellulase activity (3.546 IU/ml) at pH 7.0 and temperature 300C with yeast extract and Fpase activity (0.653 IU/ml) through solid state fermentation. In future agriculture applications and in industries the cellulase enzyme production attains a crucial ro...
Sequential cellulase production, saccharification and ethanol fermentation using rice straw
2012
This study presents alkali pretreated rice straw for cellulase production using Aspergillus niger CP1 by solid state fermentation (SSF). On 8th day, 140±2.4 IU g/l dry substrate (ds) carboxy methyl cellulose CMCase), 15±1.2 IU g/l ds filter paper activity (FPA) and 24±1.1 IU g/l ds â-glucosidase activities were noted. On extraction of 120 g material with 1 l of 0.1 mol/l citrate buffer, 5.9±0.12 IU ml/l CMCase, 0.62±0.001 IU ml/l FPA and 1.11±0.002 IU/ml â-glucosidase were obtained. When extracted enzyme was used for saccharification of 120 g fresh pretreated rice straw containing 100 g holocellulose, 45±0.2 g, sugars (50 g/l) were released in 900 ml hydrolysate to give 45% hydrolysis. Hydrolysate after yeast fermentation gave 15.6±0.05 g/l ethanol with a yield of 0.4±0.011 g/l/h and 78% fermentation efficiency. In sequential enzyme production, saccharification and fermentation studies, on 8th day of cellulase production by SSF, temperature was shifted from 28±1°C to 55°C for 36 h, ...
Study on Culture Conditions for A Cellulase Production From As Pergillus Unguis
ABSTRACT: Cellulase is a common name of enzymes which catalyze cellulolysis. Specially,cellulase is widely used in food processing, animal feed, chemicals, textile,fuel and pollution treatment.The objective of this research is to study on optimal conditions for the production of cellulase byAspergillus unguis. The study was designed as a comparative culture conditions such as carbon sources, moisture content, duration, nitrogen sources and citrate buffer content on cellulase production for Aspergillus unguis. Cellulase activity was determined by measuring the absorbance at λ = 540 nm with 3,5-DNS reagent. In optimized culture conditions, enzyme activity of Aspergillus unguis achieved 110.92U/ml in comparison with a commercial cellulase with 185.33U/ml in enzyme activity. The value of cellulase activity of Aspergillus unguis is 41% lower than commercial enzymes. However, enzyme in this study was raw enzyme and the cost of producing1 litter of this enzyme is just 1/8 that of purified ones. The enzyme activity would be increased by purification. That fact has proven the applicability of using the findings of this study to improve cellulase production.
Isolation, Optimization and Production of Cellulase by Aspergillus niger from Agricultural Waste
Cellulases are the group of hydrolytic enzymes such as endoglucanase (CMCase), exoglucanase, β β β β β-glucosidase (BGL) and FPase which are responsible for release of sugars in the bioconversion of the cellulosic biomass into a variety of value-added products. The cellulase producing fungi were isolated from various agriculture fields. Total 21 isolates were obtained on Czapek's Dox agar medium. Aspergillus niger was selected as most efficient enzyme producer by screening technique. Optimization of some nutritional and environmental factors like nitrogen source, temperature, pH and fermentation time were studied under submerged culture condition for cellulolytic enzyme production. Different agriculture waste material was used as carbon source. Maximum cellulolytic activity was observed in 4.2 pH media at 28°C after 96 hours in submerge condition. Wheat straw showed maximum activity of CMCase, exoglucanase, β β β β β-glucosidase and FPase were 8.38 IU/ml, 5.21 IU/ml, 0.30 IU/ml and 8.08 IU/ml, respectively followed by baggase.
Comparative study on cellulase production
Three strains of Aspergillus niger were retrieved from culture collection of the Department of Microbiology, University of Karachi, Karachi, Pakistan and were studied for their ability to produce cellulases. Cultivation at different temperatures and in presence of various carbon sources revealed that all the three strains produced more amounts of endoglucanase, βglucosidase and filter-paperase activities at 35 o C; carboxymethyl cellulose promotes the production of filter paperase and endoglucanase activities whereas salicin induced β-glucosidase activity. Experiments on growth and enzyme production kinetics showed that generation time and hence volumetric rate of biomass production is influenced by the carbon source used in the medium; simple carbon source, such as glucose favored the growth of all the strains. Cellulases from all the strains showed optimum activity at temperature >50 o C and under acidic range of pH, while melting temperature was 64-65 o C. These findings affirm that cellulases from A. niger are potential candidates as alternative to Trichoderma cellulases.
Cellulase production by Aspergillus niger on different natural lignocellulosic substrates
Cellulases are the group of hydrolytic enzymes Filter paperase (FPase), Carboxymethylcellulase (CMCase) and -glucosidase (BGL) and are responsible for release of sugars in the bioconversion of the lignocellulosic biomass into a variety of value - added products. The present study was aimed to examine cellulase production by Aspergillus niger on individual lignocellulosic substrates in both submerged (SmF) and solid State (SSF) Fermentations. Rice bran supported maximum enzyme yields followed by wheat bran in both fermentations. Among different combinations with rice bran at equal ratio (1:1w/w) tested, combination of rice bran and wheat bran served the best combination for production of cellulolytic enzymes. Maximum titers of FPase, CMCase and BGL obtained on this combination were 2.632, 2.478 and 2.984 U/mL in SmF and 29.81, 25.2 and 32.18 U/gDS in SSF respectively
Environmental Sustainability, 2021
The present study aimed at cellulase production from two fungal strains of Aspergillus niger and Aspergillus heteromorphus under solid-state fermentation (SSF) and submerged fermentation (SmF) conditions. The agricultural residue (rice straw) was tested for its suitability as solid support as well as a carbon source for microbial growth. The potential of fungi to produce enzyme rely on the species of the fungus, growth substrate (lignocellulosic biomass) and culturing method. In the present work, rice straw was used as a substrate for fungal growth under SSF and SmF conditions. Maximum cellulase activities as 6.4 IU/g FPase and 125 IU/g CMCase under SSF condition, and 3.8 IU/g FPase and 94 IU/g CMCase under SmF were achieved on the 5th day of incubation from A. heteromorphus . Similarly, FPase and CMCase activity for A. niger under SSF were 5.8 IU/g and 113 IU/g, respectively, while FPase activity 3.5 IU/g and CMCase activity 88 IU/g was observed under SmF condition. A. heteromorphu...
Energy & Fuels, 2014
Among the three Aspergillus spp. (A. niger, A. oryzae, and A. f umigatus) screened for cellulolytic enzyme production potential, A. niger produced cellulolytic enzyme in relatively higher concentrations than the other two isolates. Enzyme produced by all three isolates was optimally active at pH 5.0. Cellulases from A. niger and A. f umigatus were optimally active at 55°C, while the enzyme from A. oryzae showed optimum activity at 50°C. Cellulase from A. niger and A. f umigatus retained more than 80 and 70% activity, respectively, while cellulase from A. oryzae could retain only 20% activity at 55°C after 12 h. Cellulase from A. niger exhibited better stability at higher temperatures than the enzyme from the other two Aspergillus spp., showing half-life (t 1/2) of about 5 and 3 h at 70 and 80°C, respectively. Zymogram revealed multiple forms of endoglucanase, cellobiohydrolase, and β-glucosidase with molecular mass ranging between 28 and 154 kDa for cellulase from all three isolates. Hydrolysis of rice straw at 12.5% (w/v) with crude cellulase from A. niger HO resulted in fermentable sugar concentration and productivity of 66.2 g L −1 and 2.75 g L −1 h −1 , respectively, showing potential for the reported enzyme in biofuel industry.