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

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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.

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Acknowledgements

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.

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Authors and Affiliations

  1. 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, M. Nejib Marzouki & Ferid Abidi

  2. UR 1268, Biopolymères Interactions Assemblages, Equipe Fonctions et Interactions des Protéines, INRA, B. P. 71627, 44316, Nantes Cedex 3, France

    Jean-Marc Chobert, Thomas Haertlé & Ferid Abidi

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  1. Neyssene Aissaoui
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  2. Jean-Marc Chobert
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Correspondence to Neyssene Aissaoui.

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Aissaoui, N., Chobert, JM., Haertlé, T. et al. Purification and Biochemical Characterization of a Neutral Serine Protease from Trichoderma harzianum. Use in Antibacterial Peptide Production from a Fish By-Product Hydrolysate. Appl Biochem Biotechnol 182, 831–845 (2017). https://doi.org/10.1007/s12010-016-2365-4

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