PURIFICATION AND CHARACTERIZATION OF NOVEL SOLVENT STABLE BACILLUS CEREUS PI-C4 PROTEASE FROM POULTRY WASTE

Abstract

A total of sixty-four bacterial isolates producing enzyme protease were isolated and screened from soil samples obtained from industries. A new potent solvent stable and alkaline protease producing isolate PI-C4 was isolated from Ghazipur Poultry waste site which was identified to be Bacillus cereus based on 16S rDNA sequence analysis. Consensus sequence of 1398 bp of the strain PI-C4 has been deposited in GenBank with accession number KM211501. Furthermore, the PI-C4 enzyme was subjected to precipitation using ammonium sulphate (70% saturation), dialysis and was further concentrated by ion exchange chromatography which resulted in purification fold of 2.47 and 57.7% yield. The alkaline protease was found to be 46 kDa. Enzyme PI-C4 was also characterized with respect to temperature and pH and was found to be active at pH 9.0 and 450C. The protease possesses significant stability (64.7-82.2%) in the presence of surfactants tested. The alkaline protease possesses higher stability in solvent DMSO (107%) followed by acetone (91%) and isopropanol (82%). The protease could therefore be useful for various applications like enzymatic leather treatment, feather degradation and recovery of silver.