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Characterization of neutral lipases revealed the tissue-specific triacylglycerol hydrolytic activity in Nilaparvata lugens

机译:中性脂肪酶的表征揭示了 Nilaparvata lugens 中组织特异性三酰基甘油水解活性

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摘要

The lipid metabolism plays an essential role in the development and reproduction of insects, and lipases are important enzymes in lipid metabolism. In Nilaparvata lugens, an important insect pest on rice, triacylglycerol hydrolytic activities were different among tissues, with high activity in integument, ovary, and fat body, but low activity in intestine. To figure out the tissue-specific triacylglycerol hydrolytic activity, we identified 43 lipases in N. lugens. Of these 43 lipases, 23 belonged to neutral lipases, so this group was selected to perform further experiments on triacylglycerol hydrolysis. The complete motifs of catalytic triads, beta 9 loop, and lid motif, are required for the triacylglycerol hydrolytic activity in neutral lipases, which were found in some neutral lipases with high gene expression levels in integument and ovary, but not in intestine. The recombinant proteins of 3 neutral lipases with or without 3 complete motifs were obtained, and the activity determination confirmed the importance of 3 motifs. Silencing XM_022331066.1, which is highly expressed in ovary and with 3 complete motifs, significantly decreased the egg production and hatchability of N. lugens, partially through decline of the lipid metabolism. In summary, at least one-third of important motifs were incomplete in all neutral lipases with high gene expression in intestine, which could partially explain why the lipase activity in intestine was much lower than that in other tissues. The low activity to hydrolyze triacylglycerol in N. lugens intestine might be associated with its food resource and nutrient components, and the ovary-specific neutral lipases were important for N. lugens reproduction.
机译:脂质代谢在昆虫的发育和繁殖中起着至关重要的作用,脂肪酶是脂质代谢中的重要酶。在水稻的重要害虫Nilaparvata lugens中,三酰甘油水解活性在组织间存在差异,在外皮、卵巢和脂肪体中活性较高,但在肠道中活性较低。为了弄清楚组织特异性三酰基甘油水解活性,我们在 N. lugens 中鉴定了 43 种脂肪酶。在这43种脂肪酶中,有23种属于中性脂肪酶,因此选择该组进行三酰基甘油水解的进一步实验。催化三联征、β9 环和盖子基序的完整基序是中性脂肪酶中三酰基甘油水解活性所必需的,这些基序存在于一些在外皮和卵巢中基因表达水平高的中性脂肪酶中,但在肠道中则不然。得到3个中性脂肪酶的重组蛋白,有或没有3个完整基序,活性测定证实了3个基序的重要性。沉默XM_022331066.1在卵巢中高表达,具有3个完整的基序,显著降低了鲁根奈瑟菌的产蛋量和孵化率,部分原因是脂质代谢下降。综上所述,在所有肠道中基因表达较高的中性脂肪酶中,至少有三分之一的重要基序是不完整的,这可以部分解释为什么肠道中的脂肪酶活性远低于其他组织。鲁氏奈氏菌肠道中水解三酰基甘油的低活性可能与其食物资源和营养成分有关,卵巢特异性中性脂肪酶对鲁氏奈瑟菌的繁殖具有重要意义。

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