title: Salt dependent resistance against chemical denaturation of alkaline protease from a newly isolated haloalkaliphilic Bacillus sp.
abstract: Only few enzymes from haloalkaliphiles are biochemically characterized for their kinetic behaviour and stability. In view of this realization, an alkaline protease from Bacillus sp. AH-6, displaying salt-dependent resistance against chemical denaturation by Urea and Guanidium hydrochloride was investigated for denaturation and in vitro protein folding. The crude enzyme was highly resistant against urea (8 M) denaturation up to 72 h; however, on purification, it turned sensitive and got denatured within 2 h. Interestingly, the purified enzyme regained the resistance in the presence of NaCl. Effective refolding of the purified enzyme was achieved with glycerol; however, other approaches such as lower protein concentrations, rapid dilution and slow removal of the denaturant did not further add to refolding. The results are important from the viewpoint that only few enzymes from haloalkaliphilic bacteria are characterized. Since the resistance against chemical denaturation is a rare phenomenon, the findings would enrich the knowledge on protein stability and denaturation. Besides, such biocatalysts would definitely have novel applications under harsh chemical environments.
authors.affiliation: Department of Biosciences, Saurashtra University, Rajkot, Gujarat 360005, India.
authors.forename: M S
authors.initials: MS
authors.lastname: Dodia
authors.forename: H G
authors.initials: HG
authors.lastname: Bhimani
authors.forename: C M
authors.initials: CM
authors.lastname: Rawal
authors.forename: R H
authors.initials: RH
authors.lastname: Joshi
authors.forename: S P
authors.initials: SP
authors.lastname: Singh
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry*; D010450:Endopeptidases / Q000737:chemistry*; D005990:Glycerol / Q000737:chemistry*; D007700:Kinetics; D011489:Protein Denaturation; D012965:Sodium Chloride / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D012965:Sodium Chloride; D010450:Endopeptidases; C087822:alkaline protease; D005990:Glycerol