中国对虾中抗菌短肽的分离纯化与功能分析

Isolation and purification of a novel antimicrobial peptide from Penaeus chinesis

以水产病害的病原菌为目标菌,从中国对虾(Penaeus chinesis)体液中分离筛选抗菌肽,通过Sephadex G-50、RP-HPLC等分离纯化技术,得到一种抑制水产病原菌的抗菌肽PC-V-2。经初步鉴定,该抗菌肽具有广谱抗性,不仅抗鲁克氏耶尔森氏菌(Yersinia ruckeri),而且对金黄色葡萄球菌(Staphyloccus aureus)、枯草杆菌(Bacillus subttilis)等革兰氏阳性菌,大肠杆菌(Escherichia coli)等革兰氏阴性菌和白色念珠菌(Candida albicans)等真菌也有较强的抗性。另外,该抗菌肽还有一定的抑制凝血的活性,但没有检测到溶血和丝氨酸蛋白酶抑制活性。经MALDI-TOF质谱分析,其分子量为1 848 D。

It is one of the important things to find more effective biological medicaments to prevent the disease in aquaculture. The purpose of the study was to get antimicrobial peptide from Penaeus chinensis body fluid. Some pathogenic microorganisms were used to detect the activity of the antimicrobial peptides. The results showed that a novel antimicrobial peptide to Yersinia ruckeri was isolated and purified from Penaeus chinensis by Scphadex G-50 gel filtration and reverse phase high performance liquid chromatography. The molecular weight was 1 848 D determined by matrix-assisted laser desorption ionization time-of-flight （MALDI-TOF） mass spectrometry. It had characteristics of broad spectrum. It not only inhibited the growth of Gram positive bacteria such as Staphyloccus aureus and Bacillus subttilis, but also Gram negative bacteria such as Escherichia coli, especially Yersinia ruckeri. It also inhibited Candida albicans, a kind of fungi. The peptide could delay the coagulation time as anticoagulant did. No inhibitory effects on activities of serine proteases and hemolysis were observed in current studies. The study on the amino acid sequence and other characters of the peptide are needed in the future.