A report on thermostable alkaline protease producing bacteria from a terrestrial thermal spring

Five efficient protease producing moderate thermophiles were isolated from Unkeshwar (19 degrees 85' N and 78 degrees 25'E) thermal spring located in Nanded district of Maharashtra. The isolates were identified as Bacillus firmus, Bacillus aminivorans, Bacillus sterothermophilus, Bacillus brevis and Pseudomonas oleovorans using biochemical techniques. Being an efficient protease producer, Pseudomonas oleovorans was selected and further identified using 16s rRNA molecular analysis method as 'Uncultured bacterium clone APP30' and sequence was deposited in genebanlc nucleotide repository under accession number KC733812. The protease extracted and partially purified from APP30 showed remarkable caseinolytic activity at 55 degrees C temperature and pH 9.0. The extracted protease also showed stability in presence of Chloroform, Ethanol, Butanol, Hexane, Toluene and Triton X-100 at 1 % (v/v) concentration and compatibility with different commercial detergents. Efficient destaining of blood stained cotton cloth piece was also recorded by partially purified protease. Enhanced proteolytic activity was observed in presence of CaCl2, MgSO4, KCl and NaCl at 1 % (w/v) concentration. Characterization of partially purified protease was carried out. Molecular weight of partially purified protease was determined as 28 kda.