Applied Biochemistry and Biotechnology

, Volume 182, Issue 2, pp 831–845 | Cite as

Purification and Biochemical Characterization of a Neutral Serine Protease from Trichoderma harzianum. Use in Antibacterial Peptide Production from a Fish By-Product Hydrolysate

  • Neyssene AissaouiEmail author
  • Jean-Marc Chobert
  • Thomas Haertlé
  • M. Nejib Marzouki
  • Ferid Abidi


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.


Trichoderma harzianum Protease Purification Protein hydrolysate Antibacterial peptide 



This work was supported by the financial project of LIP-MB Laboratory, INSAT, Carthage University, and the Ministry of Higher Education and Scientific Research of Tunisia.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Neyssene Aissaoui
    • 1
    Email author
  • Jean-Marc Chobert
    • 2
  • Thomas Haertlé
    • 2
  • M. Nejib Marzouki
    • 1
  • Ferid Abidi
    • 1
    • 2
  1. 1.Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB), National Institute of Applied Sciences and TechnologyUniversity of CarthageTunis CedexTunisia
  2. 2.UR 1268, Biopolymères Interactions Assemblages, Equipe Fonctions et Interactions des ProtéinesINRANantes Cedex 3France

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