Two carboxylesterase genes in Plutella xylostella associated with sex pheromones and plant volatiles degradation.

Abstract

Carboxyl/cholinesterases (CCEs) are thought to play a pivotal role in the degradation of sex pheromones and plant-derived odorants in insects, but their exact biochemistry and physiological functions remain unclear. In this study, two paralogous antennae-enriched CCEs from Plutella xylostella (PxylCCE16a and 16c) were identified and functionally characterized. High-purity protein preparations of active recombinant PxylCCE16a and 16c have been obtained from Sf9 insect cells by Ni2+ affinity purification. Our results revealed that the purified recombinant PxylCCE016c is able to degrade two sex pheromone components Z9-14:Ac and Z11-16:Ac at 27.64 ± 0.79% and 24.40 ± 3.07%, respectively, while PxylCCE016a presented relatively lower activity. Additionally, a similar difference in activity was measured in plant-derived odorants. Furthermore, both CCEs displayed obvious preferences for the two sex pheromone components, especially on Z11-16:Ac (Km values are in the range 7.82-45.06 μmol L-1 ) which much lower than plant odorants (Km values are in the range 1290-4030 μmol L-1 ). Furthermore, the activity of the two newly identified CCEs is pH-dependent. The activity at pH6.5 is obviously higher than that at pH5.0. Interestingly, only PxylCCE016c can be inhibited by a common esterase inhibitor triphenyl phosphate (TPP) with LC50 of 1570 ± 520 μmol L-1 . PxylCCE16c plays a more essential role in odorant degradation than PxylCCE16a. Moreover, the current study provides novel potential pesticide targets for the notorious moth Plutella xylostella. © 2021 Society of Chemical Industry.