title: Cloning and functional expression of a chitinase cDNA from the apple leaf miner moth Lithocolletis ringoniella.
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.
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, Shanxi, 030024, China. fxjbio@163.com.
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Fan
authors.forename: Yan-Xia
authors.initials: YX
authors.lastname: Mi
authors.forename: Hui
authors.initials: H
authors.lastname: Ren
authors.forename: Chang
authors.initials: C
authors.lastname: Zhang
authors.forename: Yao
authors.initials: Y
authors.lastname: Li
authors.forename: Xiao-Xiao
authors.initials: XX
authors.lastname: Xian
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002688:Chitinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D006031:Glycosylation; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D010766:Phosphorylation; D010802:Phylogeny; D011487:Protein Conformation
chemical_list: D002688:Chitinases