PURIFIATION AND CHARACTERIZATION OF THERMOSTABLE - AMYLASE ISOLATED FROM IMMOBILIZED CELLS OF BACILLUS THURINGIENSISSANDIP BANDOPADHYAY
Calcium alginate entrapped cells of Bacillus thuringiensis exhibited better amylase production compared to free cells. The enzyme isolated from the immobilized cells was subsequently purified by ammonium sulphate precipitation, dialysis and ion exchange chromatography. SDS-PAGE of the purified protein revealed that the enzyme was a 52 KDa protein without the presence of any isozyme. The optimum pH and optimum temperature of the enzyme was 6.9 and 60oC respectively. The purified enzyme protein is quite stable at room temperature for 4 hours. Detailed study of the enzyme revealed that the enzyme is Ca2+ dependent and completely inhibited by trace amount of iodoacetate and p-Chloro Mercuro Benzoate (PCMB). However, the inhibition was partially withdrawn by 2-mercapto ethanol, which is an indication of the presence of SH group at the active centre of the enzyme.
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