Biochemical and molecular characterization of Pseudomonas aeruginosa CTM50182 
organic solvent-stable elastase

Jaouadi, Bassem; Jaouadi, Nadia Zarai; Rekik, Hatem; Naili, Belgacem; Beji, Abdelhamid; Dhouib, Abdelhafidh; Bejar, Samir

doi:10.1016/j.ijbiomac.2013.05.019

Item

ITEM ACTIONSEXPORT

Add to Basket

Local TagsRelease HistoryDetailsSummary

Released

Journal Article

Biochemical and molecular characterization of Pseudomonas aeruginosa CTM50182 organic solvent-stable elastase

MPS-Authors

/persons/resource/persons77735

Beji, Abdelhamid
Ullrich, Axel / Molecular Biology, Max Planck Institute of Biochemistry, Max Planck Society;

External Resource

No external resources are shared

Fulltext (restricted access)

There are currently no full texts shared for your IP range.

Fulltext (public)

There are no public fulltexts stored in PuRe

Supplementary Material (public)

There is no public supplementary material available

Citation

Jaouadi, B., Jaouadi, N. Z., Rekik, H., Naili, B., Beji, A., Dhouib, A., et al. (2013). Biochemical and molecular characterization of Pseudomonas aeruginosa CTM50182 organic solvent-stable elastase. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 60, 165-177. doi:10.1016/j.ijbiomac.2013.05.019.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0014-AA71-6

Abstract

An extracellular alkaline elastase was produced from Pseudomonas aeruginosa CTM50182. It was chromatographically purified using HPLC and Mono Q Sepharose column. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that the purified enzyme (called AMPP) was a monomer with a molecular mass of 33,015.18 Da. The N-terminal 29 amino acid sequence of AMPP showed high homology with those of Pseudomonas elastases. It showed optimal activity at pH 12 and 80 degrees C and was stable at a pH range of 9-12 after 120 h of incubation. Its thermoactivity and thermostability were upgraded in the presence of 5 mM Co2+. Its half-life times at 70 and 80 degrees C were 16 and 10 h, respectively. It was completely inhibited by ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), and 1,10-phenanthroline, suggesting that it belongs to the metalloprotease family. AMPP also exhibited high catalytic efficiency, organic solvent-tolerance, and hydrolysis. The lasB gene encoding AMPP was cloned, sequenced, and expressed in Escherichia coli. The biochemical properties of the extracellular purified recombinant enzyme (rAMPP) were similar to those of native AMPP. This organic solvent-stable protease could be considered a potential candidate for application as a biocatalyst in the synthesis of enzymatic peptides. (C) 2013 Elsevier B.V. All rights reserved.