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对虾(十足目:对虾科)中三酰甘油的分解代谢。

Triacylglycerol catabolism in the prawn Macrobrachium borellii (Crustacea: Palaemoniade).

机构信息

Instituto de Investigaciones Bioquímicas de La Plata, CONICET-CCT La Plata - UNLP, Universidad Nacional de La Plata, Argentina.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2011 Dec;160(4):201-7. doi: 10.1016/j.cbpb.2011.08.006. Epub 2011 Aug 26.

Abstract

While invertebrates store neutral lipids as their major energy source, little is known about triacylglycerol (TAG) class composition and their differential catabolism in aquatic arthropods. This study focuses on the composition of the main energy source and its catabolism by lipase from the midgut gland (hepatopancreas) of the crustacean Macrobrachium borellii. Silver-ion thin-layer chromatography of prawn large TAG deposit (80% of total lipids) and its subsequent fatty acid analysis by gas chromatography allowed the identification of 4 major fractions. These are composed of fatty acids of decreasing unsaturation and carbon chain length, the predominant being 18:1n-9. Fraction I, the most unsaturated one, contained mainly 20:5n-3; fraction II 18:2n-6; fraction III 18:1n-9 while the most saturated fraction contained mostly 16:0. Hepatopancreas main lipase (Mr 72 kDa) cross-reacted with polyclonal antibodies against insect lipase, was not dependent on the presence of Ca(2+) and had an optimum activity at 40°C and pH 8.0. Kinetic analysis showed a Michaelis-Menten behavior. A substrate competition assay evidenced lipase specificity following the order: 18:1n-9-TAG>PUFA-enriched-TAG>16:0-TAG different from that in vertebrates. These data indicate there is a reasonable correspondence between the fatty acid composition of TAG and the substrate specificity of lipase, which may be an important factor in determining which fatty acids are mobilized during lipolysis for oxidation in crustaceans.

摘要

虽然无脊椎动物将中性脂肪作为其主要能量来源储存,但关于三酰基甘油 (TAG) 类别的组成及其在水生节肢动物中的差异分解代谢知之甚少。本研究专注于甲壳类动物Macrobrachium borellii 中肠腺(肝胰腺)的脂肪酶的主要能量来源的组成及其分解代谢。银离子薄层色谱法分析对虾的大型 TAG 沉积物(占总脂质的 80%)及其随后的气相色谱法脂肪酸分析允许鉴定出 4 种主要的成分。这些成分由不饱和程度和碳链长度降低的脂肪酸组成,主要是 18:1n-9。最不饱和的 I 组分主要含有 20:5n-3;II 组分为 18:2n-6;III 组分为 18:1n-9,而最饱和的组分为 16:0。肝胰腺主要脂肪酶(Mr 72 kDa)与昆虫脂肪酶的多克隆抗体发生交叉反应,不依赖于 Ca(2+)的存在,在 40°C 和 pH 8.0 时具有最佳活性。动力学分析显示出米氏行为。底物竞争测定证明了脂肪酶的特异性,其顺序为:18:1n-9-TAG>富含多不饱和脂肪酸的-TAG>16:0-TAG,与脊椎动物不同。这些数据表明,TAG 的脂肪酸组成与脂肪酶的底物特异性之间存在合理的对应关系,这可能是决定甲壳动物在脂肪分解过程中哪些脂肪酸被动员用于氧化的重要因素。

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