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FUNCTIONAL CHARACTERIZATION OF AN ANTENNAL ESTERASE FROM THE NOCTUID MOTH, Spodoptera exigua

机译:夜蛾蛾夜蛾蛾中一类酯酶的功能鉴定

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摘要

Odorant-degrading esterases (ODEs) act in the fast deactivation of ester pheromone components and plant volatiles in insects. However, only few ODEs have been characterised to date. In this study, six full-length putative ODE genes (designated SexiCXE4, 5, 17, 18, 20, and 31) were cloned from the male antennae of Spodoptera exigua. The deduced amino acid sequences possessed typical characteristics of a carboxylesterase (CXE) and shared high identities with reported insect CXEs. The tissue and temporal expression patterns were investigated by quantitative real time PCR. Although all six SexiCXEs are expressed in antennae of both sexes, SexiCXE4, 17 and 20 are antennae-enriched; while SexiCXE5 and SexiCXE18 are dominantly expressed in wings, and SexiCXE31 is mainly expressed in proboscises, heads and legs. With the highly biased expression in antennae and proboscises, SexiCXE4 was selected for further functional assay. The recombinant SexiCXE4 were expressed in High-five cells and purified by a Ni2+ affinity column. SexiCXE4 has much higher enzyme activity against plant volatiles (Z)-3-hexenyl acetate and hexyl acetate than to the sex pheromone components, suggesting that it may function mostly in the degradation of the plant volatiles.
机译:臭味降解酯酶(ODE)可以使昆虫中的酯信息素成分和植物挥发物快速失活。但是,迄今为止,只有很少的ODE被表征。在这项研究中,从斜纹夜蛾的雄性触角中克隆了六个全长推定的ODE基因(命名为SexiCXE4、5、17、18、20和31)。推导的氨基酸序列具有羧酸酯酶(CXE)的典型特征,并且与报道的昆虫CXE具有高度同一性。通过定量实时PCR研究组织和时间表达模式。尽管所有六个SexiCXE都在两性的触角中表达,但SexiCXE4、17和20富含触角。而SexiCXE5和SexiCXE18主要在翅膀中表达,而SexiCXE31主要在鼻,头和腿中表达。由于触角和喙的表达高度偏向,因此选择SexiCXE4进行进一步的功能测定。重组SexiCXE4在高细胞中表达,并通过Ni2 +亲和柱纯化。 SexiCXE4对植物挥发物(Z)-3-己烯基乙酸酯和乙酸己酯的酶活性比对性信息素组分的酶活性高得多,这表明它可能主要在植物挥发物的降解中起作用。

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