title: Thermotolerant alkaline protease enzyme from Bacillus licheniformis A10: purification, characterization, effects of surfactants and organic solvents.
abstract: In this study, the extracellular thermostable alkaline protease out of A10 strain was purified 1.38-fold with 9.44% efficiency through the ammonium sulfate precipitation-dialysis and DE52 anion exchange chromatography methods. The molecular weight of the enzyme in question along with sodium dodecyl sulfate-polyacrylamide gel electrophoresis was determined to be approximately 40.55 kDa, whereas the optimum pH and temperature ratings were identified as 9.0 and 70 °C, respectively. It was seen that the enzyme had remained stable between pH 7.5-10.5 range, protecting more than 90% of its activity in the wake of 1 h incubation at 60-70 °C. It was also observed that the enzyme enhanced its activity in the presence of Mg(2+), Mn(2+), K(+), while Fe(2+), Ni(2+), Zn(2+), Ag(+ )and Co(2+ ) decreased the activity. Ca(2+), however, did not cause any change in the activity. The enzyme was seen to have been totally inhibited by phenylmethylsulfonyl fluoride, therefore, proved to be a serine alkaline protease.
authors.affiliation: a Department of Molecular Biology and Genetics , Faculty of Science, Atatürk University , Erzurum , Turkey.
authors.forename: Bahar
authors.initials: B
authors.lastname: Yilmaz
authors.affiliation: a Department of Molecular Biology and Genetics , Faculty of Science, Atatürk University , Erzurum , Turkey.
authors.forename: Mustafa Ozkan
authors.initials: MO
authors.lastname: Baltaci
authors.affiliation: a Department of Molecular Biology and Genetics , Faculty of Science, Atatürk University , Erzurum , Turkey.
authors.forename: Melda
authors.initials: M
authors.lastname: Sisecioglu
authors.affiliation: a Department of Molecular Biology and Genetics , Faculty of Science, Atatürk University , Erzurum , Turkey.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Adiguzel
mesh_terms: D000222:Adaptation, Physiological; D000469:Alkaline Phosphatase / Q000378:metabolism*; D000069976:Bacillus licheniformis / Q000201:enzymology*; D009930:Organic Chemicals / Q000378:metabolism; D012997:Solvents / Q000378:metabolism*; D013501:Surface-Active Agents / Q000378:metabolism*
chemical_list: D009930:Organic Chemicals; D012997:Solvents; D013501:Surface-Active Agents; D000469:Alkaline Phosphatase
keywords: Alkaline protease; Bacillus licheniformis; characterization; purification; thermotolerant