Characterization of a novel protease from Anoxybacillus kamchatkensis strain M1V with biotechnological interest

Abstract

: A total of 5 proteolytic thermphiles bacteria were isolated from Hammam Righa hot spring in Algeria. Strain M1V was selected as the best producer of an extracellular protease, called SAPA, and was used for further studies. Sequence analysis of the 16S rRNA gene in addition to phenotypic tests led to the placement of this organism in the genus Anoxybacillus and species of kamchatkensis . Maximal protease production was detected after 48 h of incubation at 45 °C. This SAPA protease was purified and biochemically characterized, showing optimal activity at 70 °C, pH 11, and high levels of hydrolysis, substrate specificity, and catalytic efficiency than purified and commercial proteases. The protease activity was strongly inhibited by phenylmethanesulfonyl fluoride (PMSF), and diiodopropyl fluorophosphates (DFP). SAPA has a molecular mass of 28 kDa, and the N-terminal amino acid sequence determined showed similarity to serine proteases previously described.