title: Purification and Biochemical Characterization of a Neutral Serine Protease from Trichoderma harzianum. Use in Antibacterial Peptide Production from a Fish By-Product Hydrolysate.
abstract: This study reports the purification and biochemical characterization of an extracellular neutral protease from the fungus Trichoderma harzianum. The protease (Th-Protease) was purified from the culture supernatant to homogeneity by a three-step procedure with 14.2% recovery and 9.06-fold increase in specific activity. The purified enzyme appeared as a single protein band after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a molecular mass of about 20 kDa. The optimum pH and temperature for the proteolytic activity were pH 7.0 and 40 °C, respectively. The enzyme was then investigated for its potential application in the production of antibacterial peptides. Interestingly, Scorpaena notata viscera protein hydrolysate prepared using the purified serine protease (Th-Protease) showed remarkable in vitro antibacterial activities. A peptide with a high antibacterial activity was further purified by a three-step procedure, and its sequence was identified as FPIGMGHGSRPA. The result of this study offers a promising alternative to produce natural antibacterial peptides from fish protein hydrolysate.
authors.affiliation: Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology, University of Carthage, B. P. 676, 1080, Tunis Cedex, Tunisia. neyssene.aissaoui@hotmail.fr.
authors.forename: Neyssene
authors.initials: N
authors.lastname: Aissaoui
authors.affiliation: UR 1268, Biopolymères Interactions Assemblages, Equipe Fonctions et Interactions des Protéines, INRA, B. P. 71627, 44316, Nantes Cedex 3, France.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Chobert
authors.affiliation: UR 1268, Biopolymères Interactions Assemblages, Equipe Fonctions et Interactions des Protéines, INRA, B. P. 71627, 44316, Nantes Cedex 3, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Haertlé
authors.affiliation: Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology, University of Carthage, B. P. 676, 1080, Tunis Cedex, Tunisia.
authors.forename: M Nejib
authors.initials: MN
authors.lastname: Marzouki
authors.affiliation: Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and Technology, University of Carthage, B. P. 676, 1080, Tunis Cedex, Tunisia.|UR 1268, Biopolymères Interactions Assemblages, Equipe Fonctions et Interactions des Protéines, INRA, B. P. 71627, 44316, Nantes Cedex 3, France.
authors.forename: Ferid
authors.initials: F
authors.lastname: Abidi
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry*; D029941:Fish Proteins / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D011492:Protein Hydrolysates / Q000737:chemistry*; D057057:Serine Proteases / Q000737:chemistry*; D014242:Trichoderma / Q000201:enzymology*
chemical_list: D000900:Anti-Bacterial Agents; D029941:Fish Proteins; D005656:Fungal Proteins; D010455:Peptides; D011492:Protein Hydrolysates; D057057:Serine Proteases
keywords: Antibacterial peptide; Protease; Protein hydrolysate; Purification; Trichoderma harzianum