title: Purification and characterization of manganese-dependent alkaline serine protease from Bacillus pumilus TMS55.
abstract: The purification and characterization of a Mn2+-dependent alkaline serine protease produced by Bacillus pumilus TMS55 were investigated. The enzyme was purified in three steps: concentrating the crude enzyme using ammonium sulfate precipitation, followed by gel filtration and cation-exchange chromatography. The purified protease had a molecular mass of approximately 35 kDa, was highly active over a broad pH range of 7.0 to 12.0, and remained stable over a pH range of 7.5 to 11.5. The optimum temperature for the enzyme activity was found to be 60 degreesC. PMSF and AEBSF (1 mM) significantly inhibited the protease activity, indicating that the protease is a serine protease. Mn2+ ions enhanced the activity and stability of the enzyme. In addition, the purified protease remained stable with oxidants (H2O2, 2%) and organic solvents (25%), such as benzene, hexane, and toluene. Therefore, these characteristics of the protease and its dehairing ability indicate its potential for a wide range of commercial applications.
authors.affiliation: Department of Biotechnology, Alagappa University, Karaikudi - 630 003, Tamil Nadu, India.
authors.forename: Kalibulla Syed
authors.initials: KS
authors.lastname: Ibrahim
authors.forename: Jeyaraj
authors.initials: J
authors.lastname: Muniyandi
authors.forename: Shunmugiah
authors.initials: S
authors.lastname: Karutha Pandian
mesh_terms: D001407:Bacillus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; 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; D019015:Geologic Sediments / Q000382:microbiology; D006863:Hydrogen-Ion Concentration; D008345:Manganese / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012623:Seawater / Q000382:microbiology; D057057:Serine Proteases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D008345:Manganese; D010450:Endopeptidases; D057057:Serine Proteases; C087822:alkaline protease