SILPAKORN UNIVERSITY SCIENCE AND TECHNOLOGY JOURNAL, Vol 9, No 1 (2015)

Yeast Cell Surface Display of Bacterial Chitinase as a New Approach for Biocontrol of Phytopathogenic Fungi

Siripan Limsirichaikul, Piracha Silpsrikul, Pachumporn Chanweerachai, Parichat Promsoongnearn, Kittipong Puengpan, Atsuko Niimi, Tomoo Ogi, Mitsuyoshi Ueda, Chanpen Wiwat




















Abstract


Bacterial chitinase (EC 3.2.1.14) is a potential biological pesticide, but its biochemical properties for agricultural usage need to be improved. We adopted a new approach for using chitinase as a biocontrol application by immobilization of chitinase onto a yeast cell surface. To display the enzyme on the surface of Saccharomyces cerevisiae, a bacterial chitinase gene of Bacillus circulans No.4.1, was fused with the 3’ half terminus of α-agglutinin and the glycosylphosphatidylinositol (GPI) anchored signal. The yeast surface display with chitinase hydrolyzed p-nitrophenyl-diacetyl-chitobiose up to 3- to 4-fold higher than that without chitinase on the cell surface and the enzyme activity was active for up to 20 hr. Our results demonstrated that chitinase was successfully displayed on the yeast cell surface in an active form with an antifungal activity against phytopathogenic fungi, especially Fusarium oxysporum. This new approach may provide an evident for a potential biocontrol against phytopathogenic fungi.

Key Words: Antifungal activity; Biocontrol; Fusarium oxysporum; Immobilized chitinase; Plant pathogen; Yeast surface display

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