Plant and Food Research, Palmerston North, New Zealand.
Insect Mol Biol. 2010 Feb;19(1):9-25. doi: 10.1111/j.1365-2583.2009.00924.x. Epub 2009 Nov 25.
We have identified lipase-like genes from an Epiphyas postvittana larval midgut EST library. Of the 10 pancreatic lipase family genes, six appear to encode active lipases and four encode inactive lipases, based on the presence/absence of essential catalytic residues. The four gastric lipase family genes appear to encode active proteins. Phylogenetic analysis of 54 lepidopteran pancreatic lipase proteins resolved the clade into five groups of midgut origin and a sixth of non-midgut lipases. The inactive proteins formed two separate groups with highly conserved mutations. The lepidopteran midgut lipases formed a ninth subfamily of pancreatic lipases. Eighteen insect and human gastric lipases were analysed phylogenetically with only very weak support for any groupings. Gene expression was measured in the larval midgut following feeding on five artificial diets and on apple leaves. The artificial diets contained different levels of triacylglycerol, linoleic acid and cholesterol. Significant changes in gene expression (more than 100-fold for active pancreatic lipases) were observed. All the inactive lipases were also highly expressed. The gastric lipase genes were expressed at lower levels and suppressed in larvae feeding on leaves. Together, protein motif analysis and the gene expression data suggest that, in phytophagous lepidopteran larvae, the pancreatic lipases may function in vivo as galactolipases and phospholipases whereas the gastric lipases may function as triacylglycerol hydrolases.
我们从 Epiphyas postvittana 幼虫中肠 EST 文库中鉴定出了脂肪酶样基因。在 10 个胰脂肪酶家族基因中,有 6 个似乎编码有活性的脂肪酶,4 个编码无活性的脂肪酶,这是基于必需催化残基的存在/不存在。四个胃脂肪酶家族基因似乎编码有活性的蛋白质。54 种鳞翅目胰脂肪酶蛋白的系统发育分析将该分支分为五个来源于中肠的组和第六个非中肠脂肪酶。无活性蛋白形成两个具有高度保守突变的独立组。鳞翅目中肠脂肪酶形成了胰脂肪酶的第九个亚家族。分析了 18 种昆虫和人类胃脂肪酶的系统发育,但对于任何分组都只有非常微弱的支持。在幼虫中肠中,用五种人工饲料和苹果叶喂养后测量基因表达。人工饲料中含有不同水平的三酰基甘油、亚油酸和胆固醇。观察到基因表达的显著变化(活性胰脂肪酶的变化超过 100 倍)。所有无活性的脂肪酶也高度表达。胃脂肪酶基因的表达水平较低,在以叶片为食的幼虫中受到抑制。综上所述,蛋白质结构域分析和基因表达数据表明,在植食性鳞翅目幼虫中,胰脂肪酶可能在体内作为半乳糖脂酶和磷脂酶发挥作用,而胃脂肪酶可能作为三酰基甘油水解酶发挥作用。