Purification and characterisation of a 31-kDa chitinase from the Myzus Persicae aphid: a target for hemiptera biocontrol.
Hydrolytic enzymes involved in chitin degradation are important to allow moulting during insect development. Chitinases are interesting targets to disturb growth and develop alternative strategies to control insect pests. In this work, a chitinase from the aphid Myzus persicae was purified with a 36-fold purification rate in a three step procedure by ammonium sulphate fractionation, anion-exchange chromatography on a DEAE column and on an affinity Concanavalin A column. The purified chitinase purity assessed by 1D and 2D SDS-PAGE revealed a single band and three spots at 31 kDa, respectively. Chitinases were found to have high homologies with Concanavalins A and B, two chitinase-related proteins, a fungal endochitinase and an aphid acetylhydrolase by peptide identification by Maldi-Tof-Tof. The efficiency of two potent chitinase inhibitors, namely allosamidin and psammaplin A, was tested and showed significant rate of enzymatic inhibition.
Entomologie Fonctionnelle et Evolutive, Gembloux Agro-Bio Tech, Université de Liège,Gembloux, Liège, Belgium. Frederic.Francis@ulg.ac.be
Frédéric
F
Francis
Julien
J
Saguez
Anas
A
Cherqui
Sophie
S
Vandermoten
Charles
C
Vincent
Marie-France
MF
Versali
Jacques
J
Dommès
Edwin
E
De Pauw
Philippe
P
Giordanengo
Eric
E
Haubruge
D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001042:Aphids / Q000201:enzymology*; D002688:Chitinases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000302:isolation & purification*; D004220:Disulfides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008970:Molecular Weight; D010572:Pest Control, Biological / Q000379:methods*; D014312:Trisaccharides / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology
D004220:Disulfides; D004791:Enzyme Inhibitors; D014312:Trisaccharides; C054851:allosamidin; C098378:psammaplin A; D014443:Tyrosine; D002688:Chitinases; D000117:Acetylglucosamine