牛骨胶原蛋白源抑菌肽的分离纯化及成分分析

牛骨胶原蛋白经中性蛋白酶水解,采用超滤技术和凝胶层析技术对酶解液进行分离纯化并测定对大肠杆菌的抑菌活性;借助反向高效液相色谱和基质辅助激光解析/离子化串联飞行时间质谱仪对抑菌肽进行分析.结果表明:经超滤分离后,分子质量小于10kD的组分对大肠杆菌的抑菌活性强;Sephadex G-25柱分离得到的峰Ⅰ和峰Ⅳ组分有较强的抑菌活性.经反向高效液相色谱和基质辅助激光解析/离子化串联飞行时间质谱仪分析,峰Ⅰ组分中含有的多肽种类多,分子质量集中在850～1550D之间.峰Ⅳ组分含有的多肽种类较少,分子质量集中在700～900D之间.

Isolation,Purification and Compositional Analysis of Antibacterial Peptides from Bovine Bone Collagen

Bovine bone collagen was hydrolyzed by neutral protease, and then the hydrolyate was separated and purified using ultrafiltration and gel chromatography. The antibacterial peptides obtained were analyzed by reverse high-performance liquid chromatography （RP-HPLC） and matrix-assisted laser desorption/ionization cascade time of flight mass spectrometry （MALDI-TOF-TOF-MS）. The results showed that the ultrafiltration fractions with molecular weight less than 10 kD had strong antibacterial activity against Escherichia coli. The peaks I and IV isolated by Sephadex G-25 gel chromatography bad strong antibacterial activity. By RP-HPLC and MALDI-TOF-TOF-MS analysis, the peak I contained a variety of peptides with molecular weight between 850 and 1550 D, whereas the peak IV contained fewer peptides between 700 and 900 D.