Biochemical characterization of a detergent-stable serine alkaline protease from Caldicoprobacter guelmensis.
Caldicoprobacter guelmensis isolated from the hydrothermal hot spring of Guelma (Algeria) produced high amounts of extracellular thermostable serine alkaline protease (called SAPCG) (23,000U/mL). The latter was purified by ammonium sulphate precipitation, UNO Q-6 FPLC and Zorbex PSM 300 HPLC, and submitted to biochemical characterization assays. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that the purified enzyme was a monomer, with a molecular mass of 55,824.19Da. The 19 N-terminal residue sequence of SAPCG showed high homology with those of microbial proteases. The enzyme was completely inhibited by phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DFP), which suggested its belonging to the serine protease family. It showed optimum protease activity at pH 10 and 70°C with casein as a substrate. The thermoactivity and thermostability of SAPCG were enhanced in the presence of 2mM Ca(2+). Its half-life times at 80 and 90°C were 180 and 60min, respectively. Interestingly, the SAPCG protease exhibited significant compatibility with iSiS and Persil, and wash performance analysis revealed that it could remove blood-stains effectively. Overall, SAPCG displayed a number of attractive properties that make it a promising candidate for future applications as an additive in detergent formulations.
Laboratory of Cellular and Molecular Biology, Microbiology Team, Faculty of Biological Sciences, University of Sciences and Technology of Houari Boumediene (USTHB), PO Box 32, El Alia, Bab Ezzouar, 16111 Algiers, Algeria; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of S University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia; Aix Marseille Université, IRD, Université de Toulon, CNRS, Mediterranean Institute of Oceanography (MIO), UM 110, 163 Avenue of Luminy, 13288 Marseille Cedex 9, France.
Khelifa
K
Bouacem
Laboratory of Cellular and Molecular Biology, Microbiology Team, Faculty of Biological Sciences, University of Sciences and Technology of Houari Boumediene (USTHB), PO Box 32, El Alia, Bab Ezzouar, 16111 Algiers, Algeria; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of S University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Amel
A
Bouanane-Darenfed
Laboratory of Chemical Engineering, Department of Process Engineering, Faculty of Technology, University of Blida 1, Road of Soumaâ, PO Box 270, 09000 Blida, Algeria.
Hassiba
H
Laribi-Habchi
Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of S University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Mouna Ben
MB
Elhoul
Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of S University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Aïda
A
Hmida-Sayari
Laboratory of Cellular and Molecular Biology, Microbiology Team, Faculty of Biological Sciences, University of Sciences and Technology of Houari Boumediene (USTHB), PO Box 32, El Alia, Bab Ezzouar, 16111 Algiers, Algeria.
Hocine
H
Hacene
Aix Marseille Université, IRD, Université de Toulon, CNRS, Mediterranean Institute of Oceanography (MIO), UM 110, 163 Avenue of Luminy, 13288 Marseille Cedex 9, France.
Bernard
B
Ollivier
Aix Marseille Université, IRD, Université de Toulon, CNRS, Mediterranean Institute of Oceanography (MIO), UM 110, 163 Avenue of Luminy, 13288 Marseille Cedex 9, France. Electronic address: marie-laure.fardeau@univ-amu.fr.
Marie-Laure
ML
Fardeau
Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of S University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia. Electronic address: bassem.jaouadi@cbs.rnrt.tn.
Bassem
B
Jaouadi
Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of S University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Samir
S
Bejar
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D000068016:Eubacteriales / Q000201:enzymology*; D003902:Detergents / Q000494:pharmacology*; D010450:Endopeptidases / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000737:chemistry; D007700:Kinetics; D008670:Metals / Q000737:chemistry; D057057:Serine Proteases / Q000737:chemistry*; D013379:Substrate Specificity; D013696:Temperature; D013816:Thermodynamics
D001426:Bacterial Proteins; D003902:Detergents; D004791:Enzyme Inhibitors; D007477:Ions; D008670:Metals; D010450:Endopeptidases; D057057:Serine Proteases; C087822:alkaline protease
Caldicoprobacter guelmensis; Detergent formulations; Protease