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三种鳞翅目幼虫(大蜡螟、家蚕、甘薯天蛾)血淋巴中溶菌酶特性的比较研究

Comparative study on characteristics of lysozymes from the hemolymph of three lepidopteran larvae, Galleria mellonella, Bombyx mori, Agrius convolvuli.

作者信息

Yu Kyung Hyun, Kim Kyu Nam, Lee Joon Ha, Lee Heui Sam, Kim Sang Hyun, Cho Kyung Yeon, Nam Myung Hee, Lee In Hee

机构信息

Department of Life Science, Hoseo University, 29-1 Sechuli, Baebang-Myun, Asan City Choongnam-Do 336-795, South Korea.

出版信息

Dev Comp Immunol. 2002 Oct;26(8):707-13. doi: 10.1016/s0145-305x(02)00027-7.

Abstract

Lysozymes were purified from the hemolymph of three immunized Lepidopteran larvae, Galleria mellonella, Bombyx mori, Agrius convolvuli to compare their physico-chemical properties and antibacterial activities with those of chicken lysozyme. Four lysozymes including the one from chicken had similar molecular masses and chromatographic behavior on reverse phase-high pressure liquid chromatography. Western blotting analysis using an antibody raised against G. mellonella revealed that lysozyme cross-reacted with two other insect lysozymes but not with commercial chicken lysozyme. Antibacterial activities of lysozymes were measured in two types of tests: radial diffusion assay and colony count assay. Our antibacterial tests revealed that all lysozymes have strong activities against Gram-positive bacteria and three insect lysozymes still retain a little potency against Gram-negative bacteria, while chicken lysozyme has no activity against Gram-negative bacteria. Taken together, we conclude three Lepidopteran lysozymes have a common distinct structure and have an antibacterial activity, which is absent in chicken lysozyme, against Gram-negative bacteria.

摘要

从三种免疫的鳞翅目幼虫(大蜡螟、家蚕、甘薯天蛾)的血淋巴中纯化溶菌酶,以比较它们与鸡溶菌酶的物理化学性质和抗菌活性。包括来自鸡的溶菌酶在内的四种溶菌酶在反相高压液相色谱上具有相似的分子量和色谱行为。使用针对大蜡螟产生的抗体进行的蛋白质免疫印迹分析表明,溶菌酶与其他两种昆虫溶菌酶发生交叉反应,但与市售鸡溶菌酶不发生交叉反应。通过两种测试来测定溶菌酶的抗菌活性:径向扩散试验和菌落计数试验。我们的抗菌测试表明,所有溶菌酶对革兰氏阳性菌都有很强的活性,三种昆虫溶菌酶对革兰氏阴性菌仍保留一点效力,而鸡溶菌酶对革兰氏阴性菌没有活性。综上所述,我们得出结论,三种鳞翅目溶菌酶具有共同的独特结构,并且具有鸡溶菌酶所没有的针对革兰氏阴性菌的抗菌活性。

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