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Isolation and partial characterization of a thermostable extracellular protease of Bacillus polymyxa B-17
1Department of Microbiology, College of Basic Sciences, Himachal Pradesh Agricultural University, Palampur, India.
Abstract:
Bacillus polymyxa B-17, a sporeforming psychrotroph produced a thermostable protease. The protease was purified to homogeneity from cell free broth culture by precipitation with ammonium sulfate and gel filtration through Sephadex G-100. The enzyme had a temperature optimum at 50 degrees C and shared significant activity at 70 degrees C. The protease was also active over a wide range of pH, 5.5 to 10.0, and had optimum activity at pH 7.5. It was inhibited by metal chelating agents and has a molecular weight of 30 kDa.
Insights
A novel thermostable protease was isolated from Bacillus polymyxa B-17. This enzyme exhibits optimal activity at 50°C and a broad pH range, making it suitable for various industrial applications.
Area of Science:
- Microbiology
- Enzymology
- Biochemistry
Background:
- Bacillus polymyxa B-17 is a spore-forming psychrotrophic bacterium.
- Psychrotrophic bacteria can produce enzymes active at low temperatures, but thermostable enzymes are also of industrial interest.
Purpose of the Study:
- To isolate and characterize a thermostable protease from Bacillus polymyxa B-17.
- To determine the enzyme's optimal conditions and properties for potential biotechnological applications.
Main Methods:
- Protease purification using ammonium sulfate precipitation and Sephadex G-100 gel filtration.
- Enzyme activity assays across a range of temperatures and pH levels.
- Determination of molecular weight and inhibition by metal chelating agents.
Main Results:
- A homogeneous thermostable protease was successfully purified.
- The enzyme showed optimal activity at 50°C, with significant activity at 70°C.
- Optimal activity was observed at pH 7.5, with activity spanning pH 5.5 to 10.0.
- The protease has a molecular weight of 30 kDa and is inhibited by metal chelating agents.
Conclusions:
- Bacillus polymyxa B-17 produces a robust, thermostable protease with a wide pH activity range.
- The characterized protease possesses properties suitable for various industrial processes requiring heat and pH stability.
- Further research could explore its specific applications in industries like food processing or detergents.