Aminoacylase efficiently hydrolyses fatty acid amino acid conjugates of Helicoverpa armigera potentially to increase the pool of glutamine

IF 3.2 2区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Gopal S. Kallure , Shubhranshu Shekhar Sahoo , Rutuja S. Kale , Vitthal T. Barvkar , Ravindar Kontham , Ashok P. Giri
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Abstract

One of the most prevalent bioactive molecules present in the oral secretion (OS) of lepidopteran insects is fatty acid amino acid conjugates (FACs). Insect dietary components have influence on the synthesis and retaining the pool of FACs in the OS. We noted differential and diet-specific accumulation of FACs in the OS of Helicoverpa armigera by using Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry. Interestingly, we identified FACs hydrolyzing enzyme aminoacylase (HaACY) in the OS of H. armigera through proteomic analysis. Next, we have cloned, expressed, and purified active recombinant HaACY in the bacterial system. Recombinant HaACY hydrolyzes all the six identified FACs in the OS of H. armigera larvae fed on host and non-host plants and releases respective fatty acid and glutamine. In these six FACs, fatty acid moieties vary while amino acid glutamine was common. Glutamine obtained upon hydrolysis of FACs by HaACY might serve as an amino acid pool for insect growth and development. To understand the substrate choices of HaACY, we chemically synthesized, purified, and characterized all the six FACs. Interestingly, rHaACY also shows hydrolysis of synthetic FACs into respective fatty acid and glutamine. Our results underline the importance of diet on accumulation of FACs and role of aminoacylase(s) in regulating the level of FACs and glutamine.

Abstract Image

Abstract Image

AminoacWylase 能有效水解 Helicoverpa armigera 的脂肪酸氨基酸共轭物,从而增加谷氨酰胺的含量。
鳞翅目昆虫口腔分泌物(OS)中最常见的生物活性分子之一是脂肪酸氨基酸共轭物(FACs)。昆虫的食物成分会影响口腔分泌物中脂肪酸氨基酸共轭物的合成和保留。我们利用液相色谱-四极杆飞行时间质谱法注意到,FACs 在 Helicoverpa armigera 的操作系统中的积累存在差异,且具有饮食特异性。有趣的是,通过蛋白质组分析,我们在胡蜂OS中发现了FACs水解酶氨基酰化酶(HaACY)。接下来,我们在细菌系统中克隆、表达和纯化了活性重组 rHaACY。重组的 HaACY 能水解以寄主植物和非寄主植物为食的 H. armigera 幼虫操作系统中所有六种已确定的 FACs,并释放出相应的脂肪酸和谷氨酰胺。在这六种 FAC 中,脂肪酸分子各不相同,而谷氨酰胺是常见的氨基酸。HaACY 水解 FACs 后得到的谷氨酰胺可能是昆虫生长发育的氨基酸库。为了了解 HaACY 的底物选择,我们对所有六种 FAC 进行了化学合成、纯化和表征。有趣的是,rHaACY 还能将合成的 FACs 水解为相应的脂肪酸和谷氨酰胺。我们的研究结果强调了饮食对 FACs 积累的重要性,以及氨基酰化酶在调节 FACs 和谷氨酰胺水平中的作用。
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来源期刊
CiteScore
7.40
自引率
5.30%
发文量
105
审稿时长
40 days
期刊介绍: This international journal publishes original contributions and mini-reviews in the fields of insect biochemistry and insect molecular biology. Main areas of interest are neurochemistry, hormone and pheromone biochemistry, enzymes and metabolism, hormone action and gene regulation, gene characterization and structure, pharmacology, immunology and cell and tissue culture. Papers on the biochemistry and molecular biology of other groups of arthropods are published if of general interest to the readership. Technique papers will be considered for publication if they significantly advance the field of insect biochemistry and molecular biology in the opinion of the Editors and Editorial Board.
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