Purification and Characterisation of a 31-kDa Chitinase from the Myzus Persicae Aphid: A Target for Hemiptera Biocontrol

Abstract

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 Appl Biochem Biotechnol (2012) 166:1291-1300 DOI 10.1007/s12010-011-9517-

Purification and Characterization of Trehalase From Acyrthosiphon pisum, a Target for Pest Control

“…Whilst the inhibition of chitinase was shown relevant to control harmful insects [13] as it allows for moulting during insect development, Treh only recently received attention as a promising solution for pest control. Indeed, Treh inhibitors such as validamycin A, castanospermine and trehazolin are able to disturb the regulation of trehalose catabolism, causing severe damage such as abnormal development, hampered growth, weight loss, reduced chitin synthesis, decreased flight capacity, lethal metamorphosis, unsuccessful stress recovery, as well as hypoglycaemia [3,[14][15][16].…”

Construction of an immobilised acetylcholinesterase column and its application in screening insecticidal constituents from Magnolia officinalis

“…Target enzymes are extremely important in research and development of new pesticides . The most active pesticides target the crucial metabolic enzymes for insect metabolism . Acetylcholinesterase (AChE) is such a crucial enzyme, involved in neural transmission of insects and therefore a target of various neurotoxicants.…”

Purification of bacteriophage lambda repressor

“…In this work, an efficient strategy was developed to efficiently produce tag-free, full length CI protein. Since individual proteins often exhibit unique combinations of affinity and ion exchange equilibria, even quite similar proteins may be separated on these principles (31). In the case of the CI protein, ion exchange on Q sepharose followed by heparin affinity chromatography was selected for the purification from over-expressing clones.…”