Isolation and characterization of extracellular cellulase using Bacilluls subtilis from mangrove soil (original) (raw)
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Enzymes plays a major role in industries like textile, leather, sugar, paper, rubber, tea etc., Microbes plays important role in enzyme production due to their fast growth rate, easy to manipulate for getting highly stable enzymes through genetic engineering and requires shorter time for production and purification steps. In our present study, we concentrated on the enzymes used in textile industries. Recently, many novel enzymes that prove to be the efficient in the process of desizing, peroxide removal, bio polishing have been developed for textile industries. In commercial point of view, finding of new enzymes with high activity and desired properties that can be potentially used for those processes is a continuous process. Hence, we tried to isolate the cellulolytic bacteria from red soil and produced the enzyme cellulase using specific media. The isolated bacterial species was identified as Bacillus species by morphological and biochemical analysis. Further the cellulase activity was analyzed and the parameters like pH, temperature, substrate concentration, and incubation time were also optimized.
Isolation, partial purification and characterization of cellulase from Bacillus isolates
The cellulolytic activity of Bacillus subtilis and Bacillus cereus isolated from swampy, marshy soil was studied. The enzymatic assay was carried out using Carboxymethylcellulose (CMC) and Whatman filter paper as substrate. The purified enzyme from the isolates possessed a hydrolyzing activity on both the substrates but comparatively higher fold of enzyme purification was observed in B. cereus than B. subtilis. However, CMCase activity of both the isolates was observed to be relatively higher as compared to filter paper hydrolyzing activity. The maximum fold of purification was obtained in ion-exchange chromatography than the other techniques. The optimum temperature and pH for CMCase and filter paper hydrolyzing activity were observed to be 550C and 5.5 respectively for both the isolated species. Thus, it appeared that both Bacillus subtilis and Bacillus cereus showed both endoglucanase and exoglucanase activity, as it could degrade crystalline form of cellulose.
Journal of Microbiology, Biotechnology and Food Sciences, 2013
The bioconversion of cellulose is important for global stabilization and a sustainable human society. In this study, efficient cellulolytic bacteria were screened and isolated from decayed lignocellulosic waste. Among the isolates, three strains, NASCB-5 , NASCB-8 and NASCB-12 showing higher potential for cellulase production were purified and sub cultured on carboxymethyl cellulose (CMC) agar plates. Among the three isolates NASCB-5 had given maximum enzyme activity.The isolates NASCB-5 were identified as Bacillus subtilis strains by morphological, physiological, biochemical and 16S rRNA gene analysis. The isolated strain produced cellulase enzymes complex, which suggested synergic cellulolytic systems in Bacillus subtilis. Cellulase was produced in cellulase enzyme production media containig waste paper as Carbon source and the culture conditions like temperature, pH, and Incubation time and medium components nitrogen sources were optimized. Optimal concentration of inoculum for e...
Cellulase is a hydrolytic enzyme which breaks down cellulose into smaller oligosaccharides and glucose. A total of 100 samples were collected from different sites of Allahabad, of which 40 gave positive result for cellulase production. The positive isolates were screened and selected on the basis of maximum hydrolytic zone formation on CMC agar media. The organism that gave maximum cellulase production was identified as Bacillus cereus, Bacillus Pantothenticum, Bacillus Insolitus, Bacillus alvei, Bacillus Firmus, Bacillus alvei, Bacillus Alcolphilus, Bacillus Alvei and Bacillus subtilus. Cellulase was produced by Bacillus cereus with sugarcane bagasse as solid substrate. The different parameters for cellulase production were evaluated and it was observed that the maximum production was found at 72hrs incubation period, 30°C, 7.5pH, 1%lactose additional carbon source, 1%peptone additional nitrogen source. Crude enzyme was partially purified by ammonium sulphate precipitation, dialysis and DEAE-cellulose ion-exchange chromatography. The total activity of crude enzyme and total activity of chromatography enzyme showed a 4537.2 and 323.52 respectively. A total protein content of crude and total protein content of ion-exchange chromatography showed a 1633.17mg/ml and 77.26mg/ml. DEAE-cellulose chromatography resulted in purification fold is 1.50. The effect of pH and temperature on cellulase activity was characterized, enzymatic activities were found at7.0 pH (5.86U/ml) and at 50°C (40.12U/ml). Compatibility of cellulase enzyme with different detergent brands in which maximum compatibility were examined for Tide detergent (enzyme activity 81.78U/ml).]
Cellulase Production by Bacillus subtilis isolated from Cow Dung
Cellulases are the enzymes hydrolyzing cellulosic biomass and are produced by the microorganisms that grown over cellulosic matters. Bacterial cellulases possess more advantages when compared to the cellulases from other sources. Cellulase producing bacteria was isolated from Cow dung. The organism was identified using 16 SrDNA sequencing and BLAST search. Cellulase was produced and the culture conditions like temperature, pH, and Incubation time and medium components like Carbon sources, nitrogen sources and role of natural substrates were optimized. The enzyme was further purified using ethanol precipitation and chromatography. Cellulase was then characterized using SDS-PAGE analysis and Zymographic Studies. The application of Cellulase in Biostoning was then analyzed.
Advances in Bioscience and Biotechnology, 2016
The properties of extracellular cellulase obtained from Bacillus species (FIIRO Strain B223) on supplying cellulose from Brewers Spent Grain (BSG) were studied. Generally, a crude cellulase activity between 11,757 Units to 13,930 Units was observed for both sources of carbon (BSG and crystalline cellulose). The optimum pH and temperature of 5.0, and 30˚C-40˚C for both sources of carbon was observed. Heavy metals such as copper, zinc, and iron inhibited the activities of B223 cellulase while the cellulase activities on alkaline earth metal (manganese) were moderate. The co-factor behaviour of manganese ion was also established. Local production of cellulase in Nigeria using local raw materials such as Brewers Spent Grain reduces enzyme cost; cost of finished products and increases gross domestic products.
Advances in Bioscience and Biotechnology, 2016
The properties of extracellular cellulase obtained from Bacillus species (FIIRO Strain B223) on supplying cellulose from Brewers Spent Grain (BSG) were studied. Generally, a crude cellulase activity between 11,757 Units to 13,930 Units was observed for both sources of carbon (BSG and crystalline cellulose). The optimum pH and temperature of 5.0, and 30˚C-40˚C for both sources of carbon was observed. Heavy metals such as copper, zinc, and iron inhibited the activities of B223 cellulase while the cellulase activities on alkaline earth metal (manganese) were moderate. The co-factor behaviour of manganese ion was also established. Local production of cellulase in Nigeria using local raw materials such as Brewers Spent Grain reduces enzyme cost; cost of finished products and increases gross domestic products.
2021
Cellulase is one of the important groups of enzyme for many industrial applications. It is applied in various processes of food industries, paper and pulp industry, fabric industry, medicine and pharma industry, agriculture industries etc. Microorganisms are among the most potential source for large scale production of this enzyme. The current study is aimed to isolate microorganisms which are capable of producing cellulase and to determine its efficiency for production of cellulase with different raw materials. For this study, bacteria microorganisms were isolated on CMC containing media. Screening was done using congo red (0.1%) and NaCl (1%) solution. Among various isolated species, Bacillus amyloliquefaciens was identified as the most potential strain and hence selected for further study, various raw materials which are rich in cellulose were used for cellulase production. These raw materials were sugarcane bagasse, paper pulp, molasses, orange peel, wheat bran, cassava waste, t...
Towards Excellence, 2021
Cellulolytic waste is superfluous in lithosphere and its conversion is one of the basic necessities. In current research, we have screen out potential bacterial isolates that were found capable for degradation of cellulose material. They can easily convert it into simple usable form of sugar. All isolates were capable to use cellulose in natural environmental condition such as pH, humidity and moderate temperature range. Isolation was done using to carboxymethycellulose agar medium. While, the isolates were cultivated by submerged fermentation method (Smf). Among all isolates Bacullis pumilus was found most potent strain for cellulose degradation and for production of cellulase at large scale