菲律宾蛤仔中大防御素的纯化及其抗菌活性

[Purification of a big defensin from Ruditapes philippinesis and its antibacterial activity].

作者信息

Wei Yu-Xi, Guo Dao-Sen, Li Rong-Gui, Chen Hao-Wen, Chen Pei-Xun

机构信息

Department of Biology, Qingdao University, Qingdao 266071, China.

出版信息

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Dec;35(12):1145-8.

PMID:14673509

Abstract

A novel big defensin was isolated and characterized from the plasma of Ruditapes philippinesis. It was purified to homogeneity by means of precipitation with (NH(4))(2)SO(4), gel-exclusion chromatography, two kinds of cation-exchange chromatography and named RPD-1. Its relative molecular mass was 24.8 kD by means of SDS-PAGE. By means of ABI437 amino acid sequence analyser, its 11 NH(2)-terminal amino acid sequence is AVPDVAFNAYG. Two databank systems (NCBI and EBI/EMBL) was indexed, no sequence with homology to RPD-1 was found. Furthermore, it exhibited strong inhibition on the growth of Gram-negative and -positive bacteria. Minimal inhibitory concentration (MIC) of RPD-1 against Staphylococcus aureus, Bacillus subtilis, Micrococcus tetragenus, Escherichia coli, Vibrio parahaemolyticus, Vibrio anguillarum was 9.6 mg/L, 76.8 mg/L, 38.4 mg/L, 76.8 mg/L, 19.2 mg/L, 19.2 mg/L respectively.

摘要

从菲律宾蛤仔血浆中分离并鉴定出一种新型大防御素。通过硫酸铵沉淀、凝胶排阻色谱、两种阳离子交换色谱将其纯化至同质，并命名为RPD-1。通过SDS-PAGE测得其相对分子质量为24.8 kD。利用ABI437氨基酸序列分析仪，测得其11个氨基末端氨基酸序列为AVPDVAFNAYG。检索两个数据库系统（NCBI和EBI/EMBL），未发现与RPD-1具有同源性的序列。此外，它对革兰氏阴性菌和阳性菌的生长均表现出强烈抑制作用。RPD-1对金黄色葡萄球菌、枯草芽孢杆菌、四联微球菌、大肠杆菌、副溶血性弧菌、鳗弧菌的最小抑菌浓度（MIC）分别为9.6 mg/L、76.8 mg/L、38.4 mg/L、76.8 mg/L、19.2 mg/L、19.2 mg/L。