水产学报2013，Vol.37Issue(4)：614-621,8.DOI:10.3724/SP.J.1231.2013.38284
pH和色谱柱对日本鳗鲡肝脏抗菌肽分离纯化效果的影响及抗菌活性检测
Effects of pH and chromatographic column on isolation,purification of antibacterial peptide from Japanese eel liver,and determination of antibacterial activity
摘要
Abstract
In order to quickly isolate and purify antimicrobial peptide, different flow phase pH values (cation change pH 3. 0,4. 0,5. 0, and reverse-phase liquid chromatography pH 2. 0,4. 5,7. 0), and different chromatographic columns ( cation-change and gel chromatography coupled with reverse-phase liquid chromatography respectively) were used to isolate and purify antibacterial peptide from Anguilla japonica liver,and the results were analysed and compared. The merits and drawbacks of agar plate diffusion and microporous liquid culture methods on determination of antibacterial activity were analysed. The results showed that pH 4.0 buffer solution was the best flow phase in three-pH-value buffer solutions, which had 16.43% extraction ratio and isolated two elution peaks;We did not obtain perfect result from reverse-phase liquid chromatography ( RPC ) in three-pH-value buffer solutions;Sephadex G-25 gel coupled with RPC significantly improved the isolation effect, elution peak amounts were greater, peaks were sharper and narrower,baseline was flatter and low than those of cation exchange coupled with RPC. Agar plate diffuse method to determine antibacterial activity was simple to operate, and experimental result was intuitional. It required much protein,so it should be used to determine antibacterial activity during protein crude extraction period;Microporous liquid culture method to determine antibacterial activity was sensitive. It required little protein, so it should be used to determine antibacterial activity during isolation and purification of protein in chromatography period.关键词
日本鳗鲡/抗菌肽/pH/色谱柱/抗菌活性Key words
Japanese eel/antibacterial peptide/pH/chromatographic column/antibacterial activity分类
农业科技引用本文复制引用
张东玲,关瑞章,黄文树,熊静,徐继松,宋宝东..pH和色谱柱对日本鳗鲡肝脏抗菌肽分离纯化效果的影响及抗菌活性检测[J].水产学报,2013,37(4):614-621,8.基金项目
公益性行业(农业)科研专项(nyhyzx07-043) （农业）
福建省教育厅科技项目(JA09157) （JA09157）
厦门市工程技术研究中心创新项目(3502Z20110014) （3502Z20110014）
厦门市科技局重点项目(3502Z20103023) （3502Z20103023）
