PURIFICATION AND CHARACTERIZATION OF PROTEASE PRODUCED BY BACILLUS SUBTILIS MD2KUSUMA DORCAS AND PAVAN KUMAR PINDIAbstract Bacillus subtilis MD2 (NCBI Accession no KX784212), a soil isolate was grown in casein agar and later shake flask method was used for protease production. The enzyme was purified using ammonium sulphate precipitation followed by dialysis and further concentrated using ion exchange chromatography. The SDS PAGE revealed molecular weight of the protease to be 32 kDa. The specific activity of purified enzyme was found to be 2.21 U/mg and 1.90 fold purity. The enzyme yield was 41.52% from crude protein. The optimum pH was 8.0, while the optimum temperature was 37 °C. The enzyme showed maximum activity with 1.2 % casein as substrate. It also showed increase in activity with 0.1M Fe2+, while its activity was inhibited by EDTA. 1mM of Mg2+ metal ion showed maximum protease activity.
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