title: Characterization of a novel protease from Aeribacillus pallidus strain VP3 with potential biotechnological interest.
abstract: The present study investigates the purification and physico-chemical characterization of an extracellular protease from the Aeribacillus pallidus strain VP3 previously isolated from a geothermal oil-field (Sfax, Tunisia). The maximum protease activity recorded after 22h of incubation at 45°C was 3000U/ml. Pure enzyme, designated as SPVP, was obtained after ammonium sulfate fractionation (40-60%)-dialysis followed by heat-treatment (70°C for 30min) and UNO Q-6 FPLC anion-exchange chromatography. The purified enzyme is a monomer of molecular mass about 29kDa. The sequence of the 25 NH2-terminal residues of SPVP showed a high homology with those of Bacillus proteases. The almost complete inhibition by PMSF and DIFP confirmed that SPVP is a member of serine protease family. Its optima of pH and temperature were pH 10 and 60°C, respectively. Its half-life times at 70 and 80°C were 8 and 4h, respectively. Its catalytic efficiency was higher than those of SAPCG, Alcalase Ultra 2.5L, and Thermolysin type X. SPVP exhibited excellent stability to detergents and wash performance analysis revealed that it could remove blood-stains effectively and high resistance against organic solvents. These properties make SPVP a potential candidate for applications in detergent formulations and non-aqueous peptide biocatalysis.
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Sondes
authors.initials: S
authors.lastname: Mechri
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Mouna
authors.initials: M
authors.lastname: Ben Elhoul Berrouina
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Maroua
authors.initials: M
authors.lastname: Omrane Benmrad
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Nadia
authors.initials: N
authors.lastname: Zaraî Jaouadi
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Hatem
authors.initials: H
authors.lastname: Rekik
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Emna
authors.initials: E
authors.lastname: Moujehed
authors.affiliation: Laboratory of Environmental Bioprocesses, LMI COSYS-Med, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Alif
authors.initials: A
authors.lastname: Chebbi
authors.affiliation: Laboratory of Environmental Bioprocesses, LMI COSYS-Med, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Sami
authors.initials: S
authors.lastname: Sayadi
authors.affiliation: Laboratory of Environmental Bioprocesses, LMI COSYS-Med, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Chamkha
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia.
authors.forename: Samir
authors.initials: S
authors.lastname: Bejar
authors.affiliation: Laboratory of Microbial Biotechnology and Engineering Enzymes, Centre of Biotechnology of Sfax, University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, Sfax 3018, Tunisia. Electronic address: bassem.jaouadi@cbs.rnrt.tn.
authors.forename: Bassem
authors.initials: B
authors.lastname: Jaouadi
mesh_terms: D000595:Amino Acid Sequence; D001406:Bacillaceae / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D055162:Biocatalysis; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D006207:Half-Life; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D059748:Proteolysis; D057057:Serine Proteases / Q000737:chemistry* / Q000302:isolation & purification; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D057057:Serine Proteases
keywords: Aeribacillus pallidus; Geothermal oil-field; Protease