Bm-iAANAT 及其在家蚕脂肪酸酰胺生物合成中的潜在作用。
Bm-iAANAT and its potential role in fatty acid amide biosynthesis in Bombyx mori.
机构信息
Department of Chemistry, University of South Florida, Tampa, FL 33620, USA.
Department of Chemistry, University of South Florida, Tampa, FL 33620, USA.
出版信息
Prostaglandins Leukot Essent Fatty Acids. 2018 Aug;135:10-17. doi: 10.1016/j.plefa.2018.06.001. Epub 2018 Jun 15.
The purpose of this research is to unravel the substrate specificity and kinetic properties of an insect arylalkylamine N-acyltransferase from Bombyx mori (Bm-iAANAT) and to determine if this enzyme will catalyze the formation of long chain N-acylarylalkylamides in vitro. However, the determination of substrates and products for Bm-iAANAT in vitro is no guarantee that these same molecules are substrates and products for the enzyme in the organism. Therefore, RT-PCR was performed to detect the Bm-iAANAT transcripts and liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QToF-MS) analysis was performed on purified lipid extracts from B. mori larvae (fourth instar, Bmi4) to determine if long chain fatty acid amides are produced in B. mori. Ultimately, we found that recombinant Bm-iAANAT will utilize long-chain acyl-CoA thioesters as substrates and identified Bm-iAANAT transcripts and long-chain fatty acid amides in Bmi4. Together, these data show Bm-iAANAT will catalyze the formation of long-chain N-acylarylalkylamides in vitro and provide evidence demonstrating that Bm-iAANAT has a role in fatty acid amide biosynthesis in B. mori, as well.
本研究旨在阐明家蚕芳基烷基胺 N-酰基转移酶（Bm-iAANAT）的底物特异性和动力学特性，并确定该酶是否能在体外催化长链 N-酰基芳基烷基酰胺的形成。然而，Bm-iAANAT 在体外的确定的底物和产物并不能保证这些相同的分子是该酶在生物体中的底物和产物。因此，我们进行了 RT-PCR 以检测 Bm-iAANAT 转录本，并对家蚕幼虫（四龄，Bmi4）纯化的脂质提取物进行了液相色谱四极杆飞行时间质谱（LC-QToF-MS）分析，以确定是否在家蚕中产生长链脂肪酸酰胺。最终，我们发现重组 Bm-iAANAT 将长链酰基辅酶 A 硫酯用作底物，并在家蚕 4 龄幼虫中鉴定出 Bm-iAANAT 转录本和长链脂肪酸酰胺。这些数据共同表明 Bm-iAANAT 能在体外催化长链 N-酰基芳基烷基酰胺的形成，并提供证据表明 Bm-iAANAT 在家蚕脂肪酸酰胺生物合成中发挥作用。
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