title: Purification and characterization of novel organic solvent tolerant 98kDa alkaline protease from isolated Stenotrophomonas maltophilia strain SK.
abstract: Ability of microorganisms to grow at alkaline pH makes them an attractive target for several industrial applications. Thus, search for new extremozyme producing microorganisms must be a continuous exercise. Hence, we isolated a potent alkaline protease producing bacteria from slaughter house soil. The morphological, biochemical and 16S rDNA gene sequencing studies revealed that the isolated bacteria is Stenotrophomonas maltophilia strain SK. Alkaline protease from S. maltophilia strain SK was purified by using ammonium sulphate precipitation and DEAE-cellulose ion exchange column chromatography. The purified enzyme was optimally active at pH 9.0 and temperature 40°C with broad substrate specificity. It was observed that the metal ions such as Ca(++), Mg(++) and Fe(+++) completely repressed the enzyme activity. The enzyme was stable in presence of various water miscible solvents like ethanol, methanol, isopropanol at 25% (v/v) concentration and less stable at 37.5% (v/v) concentration. These robust properties of enzyme might be applicable for various applications in detergent and pharmaceutical industries.
authors.affiliation: Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India.
authors.forename: Shailesh R
authors.initials: SR
authors.lastname: Waghmare
authors.affiliation: Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India.
authors.forename: Aparna A
authors.initials: AA
authors.lastname: Gurav
authors.affiliation: Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India.
authors.forename: Sonal A
authors.initials: SA
authors.lastname: Mali
authors.affiliation: Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India.
authors.forename: Naiem H
authors.initials: NH
authors.lastname: Nadaf
authors.affiliation: Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India.
authors.forename: Deepak B
authors.initials: DB
authors.lastname: Jadhav
authors.affiliation: Department of Microbiology, Shivaji University, Kolhapur, Maharashtra, India; Department of Biochemistry, Shivaji University, Kolhapur, Maharashtra, India. Electronic address: kds_biochem@unishivaji.ac.in.
authors.forename: Kailas D
authors.initials: KD
authors.lastname: Sonawane
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010450:Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D012988:Soil Microbiology; D020615:Stenotrophomonas maltophilia / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D008670:Metals; D010450:Endopeptidases; C087822:alkaline protease
keywords: Alkaline protease; Organic solvent; Stenotrophomonas maltophilia strain SK