PARTIAL PURIFICATION AND CHARACTERISATION OF COLD-ACTIVE METALLOPROTEASE BY BACILLUS SP. AP1 FROM APHARWAT PEAK, KASHMIR

Abstract

A gram-positive, rod shaped psychrotrophic and alkalotolerant bacterium, producing extracellular proteolytic
enzyme was isolated from the peak of Apharwat, Kashmir. The strain was identified as Bacillus sp via 16S rDNA
sequencing and was designated as Bacillus sp. AP1. Highest quantity of enzyme was secreted when strain was
grown for 30 hours at 20ºC and pH 9.0. Glucose and skim milk were the best source of carbon and substrate
respectively. The optimal activity of partially purified protease was recorded at pH 9.0, classifying the enzyme as
alkaline protease. Similarly, the protease was found to be low temperature active with maximum enzyme activity at
20ºC. Strong inhibition of activity by EGTA and EDTA defines the enzyme as metalloprotease; among metal ions,
Mn2+ enhanced enzyme activity. Finally, the washing test proved that enzyme could possibly be effective as an
additive for cold washing purposes.