Bacillus megaterium isolated from poultry farm soil was identified by standard biochemical tests and screened for the production of serine protease. Production of serine protease was done using 5 different medias by varying the type of amino acid added. The purification was done by salt precipitation, dialysis and DEAEcellulose ion exchange chromatography. The proline containing media obtained the highest fold purification out of the five different medias (leucine, lysine, proline, tryptophan and methionine cotaining media). The enzyme showed
an optimal activity at the temperature 37°C and the pH 6 which are known as its optimum temperature and pH respectively. The enzyme was proved as a Mn2+ dependent serine protease as it was activated by Mn2+ ions and inhibited by PMSF. The molecular weight of the enzyme was determined by SDS-PAGE technique as around 30kDa. It showed an excellent detergent activity on the blood stains and a very good stability in presence of locally available detrgents. The enzyme acted on the keratin protein of the chicken feather and showed a degrading capacity on the protein. So it was proved that the recently studied serine protease has a keratinase activity also. From these datas I conclude that the protease isolated from Bacillus megaterium is a Mn2+ dependent serine protease which has both keratinase and detergent activity.
Bacillus megaterium, Keratinase, serine protease, phenylmethanesulfonylfluoride(PMSF)
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