Biochemical and molecular characterization of a novel metalloprotease from Pseudomonas fluorescens strain TBS09.

Autor: Boulkour Touioui S; Laboratory of Natural Products Chemistry and Biomolecules (LNPC-BioM), Faculty of Sciences, University of Blida 1, Road of Soumaâ, PO Box 270, 09000 Blida, Algeria; Laboratoire de Biologie des Systèmes Microbiens (LBSM), École Normale Supérieure (ENS) de Kouba, PO Box 92 Vieux-Kouba, 16308, Alger, Algeria; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia., Zaraî Jaouadi N; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia., Bouacem K; 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., Ben Ayed R; Laboratory of Molecular and Cellular Screening Processes (LMCSP), Genomics and Bioinformatics Group, Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia., Rekik H; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia., Zenati B; National Centre for Research and Development of Fisheries and Aquaculture (CNRDPA), 11 Bd Amirouche PO Box 67, Bou Ismaïl, 42415, Tipaza, Algeria., Kourdali S; National Centre for Research and Development of Fisheries and Aquaculture (CNRDPA), 11 Bd Amirouche PO Box 67, Bou Ismaïl, 42415, Tipaza, Algeria., Boudjella H; Laboratoire de Biologie des Systèmes Microbiens (LBSM), École Normale Supérieure (ENS) de Kouba, PO Box 92 Vieux-Kouba, 16308, Alger, Algeria., Sabaou N; Laboratoire de Biologie des Systèmes Microbiens (LBSM), École Normale Supérieure (ENS) de Kouba, PO Box 92 Vieux-Kouba, 16308, Alger, Algeria., Bejar S; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia., El Hattab M; Laboratory of Natural Products Chemistry and Biomolecules (LNPC-BioM), Faculty of Sciences, University of Blida 1, Road of Soumaâ, PO Box 270, 09000 Blida, Algeria., Badis A; Laboratory of Natural Products Chemistry and Biomolecules (LNPC-BioM), Faculty of Sciences, University of Blida 1, Road of Soumaâ, PO Box 270, 09000 Blida, Algeria; National Centre for Research and Development of Fisheries and Aquaculture (CNRDPA), 11 Bd Amirouche PO Box 67, Bou Ismaïl, 42415, Tipaza, Algeria. Electronic address: badisabdelmalek@yahoo.fr., Annane R; National Centre for Research and Development of Fisheries and Aquaculture (CNRDPA), 11 Bd Amirouche PO Box 67, Bou Ismaïl, 42415, Tipaza, Algeria., Jaouadi B; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia. Electronic address: bassem.jaouadi@yahoo.fr.
Jazyk: angličtina
Zdroj: International journal of biological macromolecules [Int J Biol Macromol] 2018 Feb; Vol. 107 (Pt B), pp. 2351-2363. Date of Electronic Publication: 2017 Oct 18.
DOI: 10.1016/j.ijbiomac.2017.10.116
Abstrakt: A novel extracellular protease called MPDZ was purified and characterized from Pseudomonas fluorescens strain TBS09. The enzymatic properties of MPDZ were investigated using biochemical and biophysical methods. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that it was a monomer with a molecular mass of 50013.17Da. The NH 2 -terminal 27 amino acid sequence of MPDZ showed high homology with those of Pseudomonas-proteases of the serralysin family. MPDZ showed optimal activity at pH 7 and 60°C. It was totally inhibited by EGTA, EDTA, and 1,10-phenanthroline, suggesting its belonging to the metalloprotease family. Because of the interesting properties, the mpDZ gene encoding MPDZ was cloned, sequenced, and expressed in E. coli. The deduced amino acid sequence showed a strong homology with other Pseudomonas-metalloproteases. The highest sequence identity value (97%) was obtained with AprX from P. fluorescens strain CY091, with only 12 different amino acid residues. The physico-chemical properties of the extracellular purified recombinant enzyme (rMPDZ) were similar to those of MPDZ. Overall, MPDZ is bestowed with a number of promising biochemical properties that might give new opportunities for its biocatalytic applications. These data constitute an essential first step towards an understanding of the properties of MPDZ enzyme.
(Copyright © 2017 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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