黄粉虫β-1，3-葡聚糖识别蛋白的分离纯化及部分生物学功能

Purification and partial biological functions of β-1,3-glucan recognition protein from Tenebrio molitor(Coleoptera:Tenebrionidae)larvae

采用中性盐沉淀、凝胶层析等常规方法纯化黄粉虫Tenebrio molitor血淋巴中的β-1,3-葡聚糖识别蛋白,并对其在酚氧化酶原激活系统中的作用进行了初步的研究。结果表明:黄粉虫血淋巴的β-1,3-葡聚糖识别蛋白的分子量约为70kDa,主要分布于血浆中。纯化的β-1,3-葡聚糖识别蛋白只能特异性地识别β-1,3-葡聚糖而不能识别肽聚糖。在β-1,3-葡聚糖所诱导的酚氧化酶原的激活过程中,随着酚氧化酶原激活程度的提高,内源性β-1,3-葡聚糖识别蛋白的含量逐渐减少。抗β-1,3-葡聚糖识别蛋白多克隆抗体对黄粉虫血淋巴中由β-1,3-葡聚糖所诱导的酚氧化酶活性起抑制作用,且该抑制作用呈现一种剂量依赖性的趋势。上述结果有助于深入了解β-1,3-葡聚糖对黄粉虫血淋巴酚氧化酶原激活系统的激活作用。

One kind of β-1,3-glucan recognition protein （Tm-βGRP） was purified from Tenebrio molitor larvae by such traditional means as neutral salt precipitation and gel chromatography, etc., and its biological functions in pro-phenoloxidase activation system were primarily studied. The results showed that the molecular weight of Tm-βGRP was approximately 70 kDa, and the protein mainly existed in plasma. Purified Tm-βGRP could only specifically recognize β-1,3-glucan, but not peptidoglycan. During the process of pro-phenoloxidase activation induced by β-1,3-gluean, with the proceeding of prophenoloxidase activation, the amount of endogenous Tm-βGRP decreased significantly. The polyelonal antibody against Tm-βGRP could inhibit phenoloxidase activity in the hemolymph of T. molitor induced by β-1, 3-gluean in a dose-dependent manner. This research may help us deeply elucidate the biological function of β-1, 3-glucan in pro-phenoloxidase activation system in the hemolymph of T. molitor.