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Biochemistry (Moscow)

, Volume 80, Issue 2, pp 242–250 | Cite as

Cloning and functional expression of a chitinase cDNA from the apple leaf miner moth Lithocolletis ringoniella

  • Xiao-Jun FanEmail author
  • Yan-Xia Mi
  • Hui Ren
  • Chang Zhang
  • Yao Li
  • Xiao-Xiao Xian
Article

Abstract

Insect chitinase plays essential roles in chitin catabolism involved in digestion and molting during insect development. In the current work, we cloned a chitinase cDNA, LrCht5, from the apple leaf miner moth Lithocolletis ringoniella and characterized its amino acid sequence and protein properties. The L. ringoniella chitinase cDNA was 2136 bp in length with an open reading frame of 1737 bp that encodes a polypeptide of 579 amino acid residues with a predicted molecular mass of 64.4 kDa and pI of 5.49. The catalytic domain has several phosphorylation and glycosylation sites. The recombinant LrCht5 was expressed in Escherichia coli and the Spodoptera frugiperda cell line Sf9, and the LrCht5 expressed in insect cells exhibited chitinolytic activity. LrCht5 was most stable at pH 6.0 and 45°C. This work has potential application in the development of novel and more specific synthetic chitinase inhibitors for use as bioinsecticides.

Key words

Lithocolletis ringoniella chitinase RACE enzyme activity 

Abbreviations

LrCht

Lithocolletis ringoniella chitinase

MU

methylumbelliferone

RACE

rapid amplification of cDNA ends

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Copyright information

© Pleiades Publishing, Ltd. 2015

Authors and Affiliations

  • Xiao-Jun Fan
    • 1
    Email author
  • Yan-Xia Mi
    • 1
  • Hui Ren
    • 1
  • Chang Zhang
    • 1
  • Yao Li
    • 1
  • Xiao-Xiao Xian
    • 1
  1. 1.Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical EngineeringTaiyuan University of TechnologyTaiyuanShanxi, China

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