title: Purification and characterization of detergent stable alkaline protease from Bacillus amyloliquefaciens SP1 isolated from apple rhizosphere.
abstract: 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 V(max) 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.
authors.affiliation: Department of Basic Sciences (Microbiology Section), Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, (H.P.), India.
authors.forename: Shiwani
authors.initials: S
authors.lastname: Guleria
authors.affiliation: Department of Microbiology, DAV University, Jalandhar, Punjab, India.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Walia
authors.affiliation: Department of Basic Sciences (Microbiology Section), Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, (H.P.), India.
authors.forename: Anjali
authors.initials: A
authors.lastname: Chauhan
authors.affiliation: Department of Basic Sciences (Microbiology Section), Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, (H.P.), India.
authors.forename: Chand Karan
authors.initials: CK
authors.lastname: Shirkot
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D003902:Detergents / Q000378:metabolism*; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D027845:Malus / Q000382:microbiology; D008970:Molecular Weight; D063245:Plant Development; D058441:Rhizosphere; D012988:Soil Microbiology; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D003902:Detergents; D010450:Endopeptidases; C087822:alkaline protease
keywords: Alkaline protease; Bacillus amyloliquefaciens; Dehairing; Detergent stable; Thermodynamic parameters