Abstract
Baculoviruses are important biological control agents against insect pests. In this work, a chitinase from Apocheima cinerarius nucleopolyhedrovirus (ApciNPV) as a fusion protein was highly over-expressed in Escherichia coli M 15 strain and affinity purified on Ni Sepharose 6 Fast Flow column. The modeling structure of ApciNPV chitinase indicated the protein contains a polycystic kidney 1(PKD1) domain on N-terminal and a characteristic catalytic domain formed α/β triosephosphate isomerase (TIM) barrel fold like other family 18 glycohydrolases. The ApciNPV chitinase displayed both endo- and exo-chitinase activities using fluorescent oligosaccharides. Moreover, assessed by LC50 (50% lethal concentration) values, the ApciNPV chitinase showed insecticidal activity against Apocheima cinerarius, Spodoptera exigua, Hyphantria cunea, Helicoverpa armigera and Lymantria dispar. The results suggest that ApciNPV chitinase may offer a potential application as a new tool to control pest. Key words: Apocheima cinerarius nucleopolyhedrovirus, chitinase, recombinant protein, insecticidal activity.
Highlights
Chitinases are enzymes that degrade chitin, which have wide-range roles in biocontrol of pathogens and harmful insects (Cohen-Kupiec and Chet, 1998)
The results suggest that Apocheima cinerarius nucleopolyhedrovirus (ApciNPV) chitinase may offer a potential application as a new tool to control pest
The 1581bp orf38 gene from ApciNPV encoded a putative chitinase at amino acid level with 65% identity to Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), 62% identity to S. marcescens and 58% identity to Cydia pomonella granulovirus (CpGV)
Summary
Chitinases are enzymes that degrade chitin, which have wide-range roles in biocontrol of pathogens and harmful insects (Cohen-Kupiec and Chet, 1998). The chitinase from B. mori nucleopolyhedrovirus (BmNPV) plays an important role in delaying the cell lysis and decreasing the haemolymph turbidity and the degradation of the body in silkworm larvae (Wang et al, 2005). Apocheima cinerarius nucleopolyhedrovirus (ApciNPV) was first isolated and identified by the Institute of Forest Ecology and Environment Conservation, Chinese Academy of Forestry in 1979 (Yu and Wang; 1987; Yu and Wang, 1986). It is a member of the Baculoviridae family, Genus
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