Exploration of a Hot Spring for Thermostable Protease Producers

Abstract

Unkeshwar hot spring of Maharashtra was explored for the isolation of industrially important efficient thermostable protease producers. These isolates were identified based on their morphological characters, microscopic features, enzymatic profile and biochemical characteristics as Bacillus sphaericus APP21, Staphylococcus auricularis APP32, Microbacterium sp. APP41, and Kurthia sp. APP42 from Unkeshwar hot spring. Thermostable protease production was carried out using casein-yeast extract medium at 50 oC. Maximum specific activity of partially purified thermostable protease was recorded from Bacillus sphaericus APP21 (447.75 U/mg). Molecular weight of partially purified proteases from these isolates has been determined in the range of 14-29 KDa. These proteases have showed excellent thermostability in the range of temperature 45 to 65 oC. These proteases were catalytically active in the range of pH 7 to 9 and belonged to the class serine protease. Km and Vmax values were found in the range 5.26-8.34 mg/mL and 446-1380 U/mg respectively. The divalent cations Mg2+ and Mn2+ enhanced the catalytic efficiency of these proteases. These proteases showed excellent catalytic efficiency in presence of selected solvents, surfactants and detergents. These properties make them excellent candidates to use in detergent formulation, food processing, and in many biotechnologically important industries where proteolysis is needed at an elevated range of temperature.

Enhanced catalytic activity of Bacillus aryabhattai P1 protease by modulation with nanoactivator

“…Catalytic activity of partially purified P1 alkaline protease was assayed using various substrates by mixing 1 mL of P1 protease and 1 mL of assay buffer containing 10 mg/ml gelatin, BSA and feather meal individually and the protease activity was determined. The casein dependent protease activity was considered as a control for calculating percent relative activity [3,4,50].…”

Optimized production, characterization and application of alkaline proteases from taxonomically assessed microbial isolates from Lonar soda lake, India

“…The effect of selected protease inhibitors (Phenylmethylsulfonyl fluoride and Dithiothreitol), oxidizing agent (H 2 O 2 ), chelator (Na 2 EDTA), surfactants (Tween 20, Tween 80 and Triton X-100), solvents (Methanol, Ethanol, Acetonitrile, Acetone, Diethyl ether and Ethyl acetate), metal ions (Mg 2+ , Mn 2+ , Cu 2+ , Fe 3+ , Zn 2+ , Ba 2+ , Hg 2+ , Ca 2+ and Na + ) and laundry detergents (Ariel™, Surf excel™, Nirma™, Sasa™, Wheel™, Rin™ and Tide™) at their different concentrations on catalytic efficiency of the each partially purified alkaline protease from the selected isolates was determined by pre-incubating them in these chemicals for 1 h at room temperature and residual activities were measured under standard assay conditions. The laundry detergents were boiled for 15 min to denature the enzymes added by the manufacturer, and cooled prior to use (Bhunia et al, 2011;Maruthiah et al, 2013;Pathak and Rathod, 2014;Mokashe et al, 2015;Sathishkumar et al, 2015;Yadav et al, 2015).…”

Data on optimized production and characterization of alkaline proteases from newly isolated alkaliphiles from Lonar soda lake, India

“…CIS-24 BL ) was used to perform optimization of physicochemical parameters for maximum alkaline protease production. The instruments used to perform alkaline protease activity assay were cooling centrifuge machine ( Remi, Mumbai ) , and UV double beam spectrophotometer ( Shimadzu corporation ) Data format Raw and analyzed Experimental factors We isolated and further identified by polyphasic approach [1] , [2] , [3] , [4] , [5] , [6] , [7] , [8] , [9] , [10] , [11] , [15] five alkaline protease producers namely Brachybacterium sp. LAP214, Bacillus cohnii LAP217, Bacillus pseudofirmus LAP220, Brevibacterium casei LAP223 and Halomonas venusta LAP515 from Lonar soda lake, India.…”