Purification and characterization of detergent stable alkaline protease from Bacillus amyloliquefaciens SP1 isolated from apple rhizosphere

Journal of Basic Microbiology - Tập 56 Số 2 - Trang 138-152 - 2016
Shiwani Guleria1, Abhishek Walia2, Anjali Chauhan1, C. K. Shirkot1
1Department of Basic Sciences (Microbiology Section), Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, (H.P.), India.
2Department of Microbiology, DAV University, Jalandhar, Punjab, India

Tóm tắt

A thermostable extracellular alkaline protease producing Bacillus amyloliquefaciens SP1 was isolated from apple rhizosphere having multifarious plant growth promoting activities. Strain SP1 was purified to 6.48‐fold using four‐step purification protocol and characterized in detail for its robustness and ecofriendly application in leather and detergent industries. Structural analysis revealed that the protease was monomeric and had a molecular weight of 43 kDa. It exhibited optimum activity at 60°C in alkaline environment (pH 8.0) and stable in the presence of surfactants and oxidizing agents. Enzyme was thermostable at 50°C and retained more than 70% activity after 30 min incubation. It has shown stain removal property and dehairing of goat skin without chemical assistance and hydrolyzing fibrous proteins. This protease showed Km of 0.125 mg ml−1 and Vmax of 12820 μg ml−1 indicating its excellent affinity and catalytic role. Thermal inactivation of the pure enzyme followed first‐order kinetics. The half life of the pure enzyme at 50, 60, and 65°C was 77, 19.80, and 13.33 min, respectively. The activation energy was 37.19 KJ mol−1. The results suggest that the B. amyloliquefaciens SP1 has a potential application in different industries.

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Thông tin xuất bản

Journal of Basic Microbiology

Tập 56 Số 2

138-152

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