Molecular Characterization of Cellulase Producing Microbacterium Barkeri Isolated from Agricultural Waste (original) (raw)
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Optimization of Cellulolytic Bacteria from Cellulose Waste Materials and its Activity
Cellulases are the group of hydrolytic enzymes and they are capable of degrading all types of cellulosic waste materials. Present study focuses on the isolation, screening and enzymatic hydrolysis cellulolytic bacteria from various agricultural wastes under submerged fermentation and optimization of pH, temperature, carbon and nitrogen sources for the activity of enzyme production was tested. The enzyme reaction was carried out by CMC (Carboxy methyl cellulose) and the production was analyzed individually by Miller‘s modified method of Dinitro-salicylic acid (DNS). The cellulase producing bacteria was identified as Bacillus sp from Tapioca leaf, Pseudomonas aeroginosa from sugarcane Bagassee, Pseudomonas flourescens from Bamboo leaf, Cellulomonas sp from Sweet lemon peel, Micrococcus sp from Tapioca shell, Streptococcus sp from Turmeric leaf and Lactobacillus sp from Groundnut shell etc are the isolates shown a good results. Seven different bacterial strains were isolated and screened for cellulase production in submerged fermentation process. Among these seven tested bacterial strains; Bacillus sp secretes maximum amount of cellulose production around 7.896(IU/ml) under a submerged fermentation. The optimum pH and temperature for the activity of enzyme was 7 and 50°C and acetate as carbon source and ammonium sulfate as nitrogen source respectively. Keywords: Agricultural waste, Cellulases, Cellulase producing bacteria, CMC (carboxymethylcellulose).
Production, Characterization and Industrial Applications of Cellulase Derived from Agro-waste
Current Journal of Applied Science and Technology
Agricultural wastes are widely available in the form of crop residues (residual stalks, straw, leaves, roots, husks, sheaths, bagasse etc.). Significant amount of these by-products derived from agroprocessing industries goes to waste. Improper disposition of such agricultural waste contributes to environmental pollution and supports the growth of several microorganisms. These wastes are lignocellulosic in nature and it could be recycled or further processed to extract or formulate value added products. Utilization and disposal of such agricultural waste is therefore essential and is progressively gaining attention as we drive towards a greener environment. The present review is focused on utilization of lignocellulosic biomass for cellulases production and its applications. Cellulose is considered one of the most important sources of carbon and its annual biosynthesis occurs in many tonnes per annum. Recycling of agricultural residue can be achieved naturally and artificially by microorganisms. Aerobic organisms such as fungi, bacteria, and some anaerobic organisms have been found to have the capability to degrade some or most of the constituents of these residues. Fungi play a significant role in the degradation of cellulose under aerobic conditions. Cellulases are important enzymes not only for their valuable applications in different sectors like Mini-review Article
Cellulose from Brewers Spent Grain (BSG) and crystalline cellulose were utilized as sole carbon source for the submerged production of Cellulase. A total of 21 bacterial isolates were obtained from Brewers Spent Grain undergoing deterioration. Out of the 21 isolates, 7 bacterial isolates showed various degrees of Cellulase production on plate assay (Nutrient Agar +2% (W/V) Cellulose) by their zones of clearance of Cellulose. The Brewers Spent Grain was characterized for its composition, and its moisture content, Crude Fibre, Fat, Protein, Ash and Total carbohydrate were 17.62%, 13.94%, 8.08%, 25%, 3.68% and 31.68% respectively. Cellulase production screening of bacterial isolates showed that Bacillus species strain B223 had the highest zone of clearance (2.8cm ± 0.02), and was used for the submerged Cellulase production. Microbial population dynamics in the enzyme production medium in the 250ml capacity Erlenmeyer flasks containing BSG cellulose increased progressively from zero hour to the 120 hour. pH of the BSG cellulose medium increased from 5.28 to 6.89, while the setup containing crystalline cellulose had its pH increased from 5.28 to 7.46. Crude cellulase activity of the cellulase in Filter paper Units (FPU) for the cellulase produced from BSG Cellullose ranged from 328 FPU, 2749PU, 32166FPU, 31910 FPU, and 33130 FPU at zero hour, 24hour, 48hour, 72 hour, 96 hour and 120 hour of fermentation respectively. The optimum pH for the production of Cellulase was 6.89. Specific Cellulase activities of Cellulase produced using BSG cellulose and Crystalline cellulose were between 12.45-4319.4 (Unit/Mg) and 19.6-5711 (Unit/Mg) respectively. Cellulases are industrial enzymes with broad applications, and production using raw materials like cellulose from Brewers Spent Grain makes the enzyme cost effective, and converts the waste (BSG) to wealth. Further research actions are ongoing to purify the crude Cellulase.
Production of cellulose from cellulomonas uda using plant waste - A Value Addition Strategy
2014
The role of enzymes in a variety of applications in various Industries gave a promising business opportunity in the present Biotechnology sector. Modern Firms are using it as a Value addition strategy to improve their value of the product. It not only increases the value but also enhances its utility. In the present work Cellulomonas uda is taken as main bacterial source for the production of Cellulase enzyme. A mutant strain of ellulomonas uda was used to explore its ability to produce cellulase enzyme at those conditions which can support the production of glucosidase from the same strain. As the basic media composition and few parameters given to bacterial strain during cultivation is constant, main concentration of the work was laid on the quantitative cellulose levels exhibited by plant waste as substrate and the strain at various time intervals & various nitrogen sources.
Bangladesh Journal of Microbiology , 2016
Microbial Cellulases have an escalating demand in many industries and constitute a major group of the industrial enzymes. It has attracted the attention of many researchers because of its tremendous industrial applications including textile industry, pulp, and paper industry, laundry and detergent industry, food and animal feed industry. The present study pursues to unfold a novel cellulase that can overcome existing challenges in biorefineries as well as to reduce biofuel production cost. Therefore, soil from a dairy farm was screened for potent cellulase producers on carboxymethylcellulose agar. Out of 68 isolates, 31 expressed cellulase activity. The best isolate so far had an extracellular crude enzyme activity of 0.167 U/ml and specific activity of 0.333 U/mg. The cell morphology, cultural characteristics, and biochemical tests presumptively identified it to belong to the genus Bacillus. Molecular analysis using 16S rRNA gene of the isolate indicated it to be Bacillus subtilis. The optimum pH and temperature for the activity of the crude enzyme were determined to be 5 and 65°C respectively.
International Journal of Research in Biosciences, 2016
The cellulase producing bacteria were isolated from industrial and agricultural areas in Kerala. Potential isolates with cellulase production were identified by Grams iodine dye staining method. The isolate were tentatively identified to be Bacillus species based on cultural, morphological, biochemical analysis and labelled as CB3, CB4 and CB8. Further, the genomic DNA was isolated and amplified with universal primers 27F and 1492s specific for 16S rRNA. The amplified 16S rRNA PCR product of 1500bp was sequenced and the unknown organism was identified using the maximum aligned 16SrRNA sequences available in the GenBank of NCBI through BLAST search. The sample CB3 and CB4 showed (100% and 99% respectively) homology to Bacillus subtilis strain. The sample CB8 showed 98% homology to the Bacillus cereus strain. To test the evolutionary relationships, phylogenetic analysis was performed with the program MEGA 6.0 using the 16SrRNAsequence. The isolates were then evaluated by submerged fermentation process for maximum cellulase production. The various process parameters like pH, temperature, incubation period, substrate concentration and inoculum volume were then optimized for the maximum production of cellulase by the isolates. The optimum temperature for cellulase production for CB4, CB4 and CB8 was found to be 40 0 C, 30 0 C and 40 0 C respectively. The optimum PH was found to be 7 for the three samples. The incubation period of 48 hours, 72 hours and 96 hours were found to be optimum for CB3, CB4 and CB8 respectively. An inoculum size of 6% was found to be ideal for CB3 and CB4 whereas an inoculums volume of 8% was found to be ideal for CB8 which showed a maximum activity of 4.12U/ml. The CMC concentration of 1.5% was found to be ideal for CB3 and CB4 whereas CB8 showed a maximum activity of 3.21U/ml at a CMC concentration of 1%. Among the three isolates the Bacillus cereus strain (CB8) was found to be the most active cellulase producer with maximum activity of 4.12 IU/ml in submerged fermentation.
Study on the Cellulase Activity of Fungi by Solid State Fermentation Using Cellulosic Wastes
2016
The present study aims to use of fungi for the degradation of cellulosic wastes like Waste Paper, Cotton ginning, Wheat Bran, Sugarcane bagasse and Cellulose using solid state fermentation for cellulase production. Total 34 isolates were obtained from various sources by the primary screening technique from which 14 isolates were showing higher cellulase activity. Potential isolates were obtained from wood furnishing region and paper industry waste. Different fungal strains were tested to find their ability to produce cellulase, which catalyses the degradation of cellulose, which is a linear psolymer made of glucose subunits linked by 1-4 glyosidic bonds. Selected 14 fungi strains were inoculated on different agriculture waste for the production of cellulase enzyme. Among these, wheat bran gave maximum zone of hydrolysis of carboxy-methyl cellulose and shown higher activities of the cellulase, which were determined by Filter paper assay (FPA) and Carboxy-methly cellulase assay (CMCas...
Elsevier, 2019
Solid-state fermentation (SSF) is a very useful modern-day tool that replenishes microbial growth on a moist surface if inert carriers and insoluble substrates are used. SSF offers several proportionate monetary as well as implementation advantages over the earlier submerged fermentation (SmF) technique that yields products with higher concentration, curtailed costs, and uncomplicated fermentation equipment, and generates reduced amounts of effluents along with a waste output that is more or less negligible. In the current scenario, SSF finds its place mostly in enzyme production as well as in organic compounds and metabolite manufacturing. Biofuel, paper, and pulp, along with detergent and textile production, show great dependence on cellulases. Endoglucanase, exoglucanase, and β-glucosidase are the three major cellulose components that can efficaciously unwind the chains of cellulose into small lignocellulose subunits and thereby produce smaller cellobiose and glucose sugar units. Different types of fungal species (filamentous, thermophilic, endophytic, etc.) are the most popular cellulase producers because they can degrade the cellulase comprehensively. A wide array of fungal species are involved in the vast production of cellulase these days with the help of this strategy because it offers lower cost with higher cellulase production.