title: Purification and characterization of organic solvent stable serine alkaline protease from newly isolated Bacillus circulans M34.
abstract: A protease from newly isolated Bacillus circulans M34 was purified by Q-Sepharose anion exchange chromatography and Sepharose-bacitracin affinity chromatography followed by (NH4)2SO4 precipitation. The molecular mass of the purified enzyme was determined using SDS-PAGE. The optimum pH and temperature for protease activity were 11 and 50°C, respectively. The effect of various metal ions on protease activity was investigated. Alkaline protease from Bacillus circulans M34 wase activated by Zn(2+), Cu(2+) and Co(2+) up to 31%. The purified protease was found to be stable in the organic solvents, surfactants and oxidizing agent. The substrate specificity of purified protease was investigated towards different substrates. The protease was almost completely inhibited by the serine protease inhibitor phenylmethanesulfonyl fluoride. The kinetic parameters of the purified protease, maximum rate (Vmax) and Michaelis constant (Km), were determined using a Lineweaver-Burk plot.
authors: Ankara University, Faculty of Science Department of Chemistry, Turkey.
authors: Esma
authors: E
authors: Sari
authors: Gazi University, Faculty of Science Department of Chemistry, Turkey.
authors: Elif
authors: E
authors: Loğoğlu
authors: Ankara University, Faculty of Science Department of Chemistry, Turkey.
authors: Atilla
authors: A
authors: Öktemer
mesh_terms: D001407:Bacillus / Q000737:chemistry / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009930:Organic Chemicals / Q000737:chemistry; D057057:Serine Proteases / Q000737:chemistry* / Q000302:isolation & purification*; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D009930:Organic Chemicals; D010450:Endopeptidases; D057057:Serine Proteases; C087822:alkaline protease
keywords: Q-Sepharose anion exchange chromatography; Sepharose-bacitracin affinity chromatography; characterization; serine alkaline protease