Expression, Purification and Characterization of Chitin-binding in Protein CPT1 from Ostrinia furnacalis

doi:10.16409/j.cnki.2095-039x.2023.03.005

Abstract

Abstract: Asian corn borer, Ostrinia furnacalis, is an important agricultural insect pest that greatly impairs corn production in China. Insect cuticle plays key roles in protecting insects from locomotion, mechanical damage or environmental stresses. The cuticle proteins that belong to Tweedle (CPT) family are known to play important roles in insect cuticle formation. Investigation on CPT may help the development of new methods for insect pest control. In this study, OfCPT1, one of the CPTs derived from the Asian corn borer, was recombinantly expressed as inclusion bodies in Escherichia coli. The obtained OfCPT1 was purified by metal chelate chromatography and then renatured with a yield of 50 mg/L. Chitin binding assay showed that OfCPT1 had a strong binding ability with chitin nanofibers, but moderate binding to α-chitin, β-chitin, chitosan, colloid chitin, and chitin nanocrystal. The circular dichroism spectra of OfCPT1 before and after binding with chitosan, showed a structural change from α-helix dominance to β-sheet dominance. This work has accomplished the recombinant preparation of OfCPT1 and revealed its biochemical properties for the first time.

Key words: cuticular protein, Tweedle, chitin-binding protein, pesticide target

CLC Number:

S476

JIANG Wang, BAO Han, CHEN Jinli, LIU Tian, YANG Qing. Expression, Purification and Characterization of Chitin-binding in Protein CPT1 from Ostrinia furnacalis[J]. Chinese Journal of Biological Control, 2023, 39(2): 462-470.

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