Serra Montserrat, Piña Benjamin, Abad José Luis, Camps Francisco, Fabriàs Gemma
Research Unit on BioActive Molecules (RUBAM), Departament de Química Orgànica Biològica, Institut d'Investigacions Químiques i Ambientals de Barcelona, Consejo Superior de Investigaciones Científicas (CSIC), Barcelona, Spain.
Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16444-9. doi: 10.1073/pnas.0705385104. Epub 2007 Oct 5.
The sex pheromone of the female processionary moth, Thaumetopoea pityocampa, is a unique C16 enyne acetate that is biosynthesized from palmitic acid. Three consecutive desaturation reactions transform this saturated precursor into the triunsaturated fatty acyl intermediate: formation of (Z)-11-hexadecenoic acid, acetylenation to 11-hexadecynoic acid, and final Delta(13) desaturation to (Z)-13-hexadecen-11-ynoic acid. By using degenerate primers common to all reported insect desaturases, a single cDNA sequence was isolated from total RNA of T. pityocampa female pheromone glands. The full-length transcript of this putative desaturase was expressed in elo1Delta/ole1Delta yeast mutants (both elongase 1 and Delta(9) desaturase-deficient) for functional assays. The construct fully rescued the Deltaole1 yeast phenotype, confirming its desaturase activity. Analysis of the unsaturated products from transformed yeast extracts demonstrated that the cloned enzyme showed Delta(11) desaturase, Delta(11) acetylenase, and Delta(13) desaturase activities. Therefore, this single desaturase may account for the three desaturation steps involved in the sex pheromone biosynthetic pathway of the processionary moth.
松异舟蛾(Thaumetopoea pityocampa)雌蛾的性信息素是一种独特的C16烯炔乙酸酯,由棕榈酸生物合成。三个连续的去饱和反应将这种饱和前体转化为三不饱和脂肪酰中间体:形成(Z)-11-十六碳烯酸,乙炔化生成11-十六碳炔酸,最后进行Δ(13)去饱和生成(Z)-13-十六碳烯-11-炔酸。通过使用所有已报道昆虫去饱和酶共有的简并引物,从松异舟蛾雌蛾性信息素腺的总RNA中分离出一个单一的cDNA序列。该假定去饱和酶的全长转录本在elo1Δ/ole1Δ酵母突变体(延长酶1和Δ(9)去饱和酶均缺陷)中表达以进行功能测定。构建体完全拯救了Δole1酵母表型,证实了其去饱和酶活性。对转化酵母提取物中不饱和产物的分析表明,克隆的酶具有Δ(11)去饱和酶、Δ(11)乙炔化酶和Δ(13)去饱和酶活性。因此,这种单一的去饱和酶可能参与了松异舟蛾性信息素生物合成途径中的三个去饱和步骤。