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As the demand for proteases increases, it was expected that active strains could be used as catalysts in different industries.
Alkaline protease produces rapid lysis of viscous exudates, show good results in the treatment of inflammation foci and
edemas, accelerates and potentiates the effect of antibiotics and sulfamide preparations and decomposes viruses and bacterial
toxins. In the present work, the strain employed was Bacillus Subtilis. The genus Bacillus was erected by Cohn in 1872 to
accommodate a bacterium described by C.G. Erhenbergis 1835 as vibrio subtilis. And it had growth at 37
C, in a shaker incubator.
Cheaper sources of both carbon and nitrogen sources are the key attraction for the commercialization of the production process
and thus ability of the microbial agent to grow and produce enzymes using these sources has been arguably a point of interest
(Mehta. V & Kanekar. P.P et al). Using batch fermentor was done by the mass production of protease (72 Hrs). The optimal
conditions for enzyme production were PH-8.0(3273U/ml), citric acid and mannose as carbon sources and yeast extract as
nitrogen source along with salts. The enzyme was immobilized on calcium alginate by cell entrapment. Protease immobilization
is important in many applications, such as biosensors, bioorganic synthesis and protein hydrolysis, heterogeneous biocatalysts,
selective absorbents, controlled released protein drugs, analytical device and solid phase protein chemistry. The optimal reaction
temperature of enzyme was 160C (2967U/ml). The immobilization process significantly improved thermal and storage stability
(1hr at room temperature and at 5
c it will be up to 1-2 months) of the free enzyme. These were 5
C. Centrifugation was
done at 27
C, and productivity was improved after immobilization than free enzyme. Immobilized Enzyme activity was 1000%
more than free enzyme Activity. The Protein estimated in enzyme assay at 300nM was 47.5KDA.
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