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Protein Engineering of Chit42 Towards Improvement of Chitinase and Antifungal Activities

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Abstract

The antagonism of Trichoderma strains usually correlates with the secretion of fungal cell wall degrading enzymes such as chitinases. Chitinase Chit42 is believed to play an important role in the biocontrol activity of Trichoderma strains as a biocontrol agent against phytopathogenic fungi. Chit42 lacks a chitin-binding domain (ChBD) which is involved in its binding activity to insoluble chitin. In this study, a chimeric chitinase with improved enzyme activity was produced by fusing a ChBD from T. atroviride chitinase 18–10 to Chit42. The improved chitinase containing a ChBD displayed a 1.7-fold higher specific activity than chit42. This increase suggests that the ChBD provides a strong binding capacity to insoluble chitin. Moreover, Chit42-ChBD transformants showed higher antifungal activity towards seven phytopathogenic fungal species.

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Acknowledgments

We thank Prof. Dr. R. L. Mach and Prof. Dr. M. J. Hynes for kindly providing plasmids. We wish to thank Dr. M. C. Limon for her advises. This project was supported by the National Institute of Genetic Engineering and Biotechnology.

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Authors and Affiliations

  1. National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrak-e Pajoohesh, km 15, Tehran - Karaj Highway, P.O. Box 14965-161, Tehran, Iran

    Mojegan Kowsari, Mostafa Motallebi & Mohammadreza Zamani

  2. Agricultural Biotechnology Research Institute of Iran, Seed and Plant Improvement, Institutes Campus, Mahdasht Road, P. O. Box 31535-1897, Karaj, Iran

    Mojegan Kowsari

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  1. Mojegan Kowsari
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  2. Mostafa Motallebi
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Correspondence to Mostafa Motallebi.

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Kowsari, M., Motallebi, M. & Zamani, M. Protein Engineering of Chit42 Towards Improvement of Chitinase and Antifungal Activities. Curr Microbiol 68, 495–502 (2014). https://doi.org/10.1007/s00284-013-0494-3

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  • DOI: https://doi.org/10.1007/s00284-013-0494-3

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