枯草芽孢杆菌BSD-2一种抗菌肽的分离纯化与鉴定

Purification and Identification of an Antimicrobial Peptides from Bacillus subtilis BSD-2

为了分离纯化及鉴定枯草芽孢杆菌BSD-2代谢产物中抗菌肽,采用盐酸沉淀、Sephadex G-50层析以及反相HPLC对其进行分离纯化,并利用Tricine-SDS-PAGE、MAIDL-TOF-MS、红外光谱和核磁共振技术对其分子量、氨基酸序列以及结构进行初步解析。结果显示该抗菌肽是由29个氨基酸组成,分子量为3 514 Da,等电点为10.46。红外光谱和核磁共振分析其结构中含有-CH2-、-CH3、-OH、-NH2、-CH、-CO-以及苯环等官能团。

In order to purify and identify an Antimicrobial peptides from the metabolic products of the Bacillus subtilis BSD-2.It was purified by Hydrochloric acid precipitation,Sephadex G-50 Chromatography and RT-HPLC.And the molecular weight was determined by Tricine-SDS-PAGE and Bioautography.The amino acid sequence and structure were preliminary analysised by MAIDL-TOF-MS,Infrared spectrum and nuclear magnetic resonance.The result indicated that the antimicrobial substance was composed of 29 amino acids.The molecular weight and PI were 3 514 Da and 10.46 respectively.The structure contained a lot of functional groups by infrared spectrum and NMR,such as-CH2-,-CH3,-OH,-NH2,-CH,-CO-and so on.