牛肺泡表面活性蛋白A的分离纯化及抗菌活性分析

doi:10.11843/j.issn.0366-6964.2021.02.022

摘要/Abstract

摘要： 旨在分离纯化牛天然肺泡表面活性蛋白A （SP-A）和分析其抗菌活性，首先通过共价交联法制备出可特异吸附SP-A的麦芽糖-Sepharose （MS）胶粒，然后用MS胶粒从牛肺泡洗出液中吸附牛SP-A，从而得到牛SP-A。SDS-PAGE和Western blot检测表明，牛SP-A单体的相对分子质量为30 ku，二聚体为60 ku。分离纯化的牛SP-A能够凝聚大肠杆菌，表明SP-A具有生物活性。抗菌活性检测结果表明，牛SP-A在体外可明显抑制金黄色葡萄球菌、无乳链球菌和致病性大肠杆菌的增殖，尤其是对革兰阴性菌的抑制作用更为明显，表明本试验制备的牛天然SP-A具有抗菌活性，这为进一步研究其生物学特性及抗菌和抗病毒活性提供了必要的条件，也为SP-A作为抗菌制剂的研发提供了新的思路。

关键词: 牛表面活性蛋白A, 分离纯化, 麦芽糖-Sepharose胶粒, 抗菌活性

Abstract: To isolate natural bovine lung surfactant protein A (SP-A) and analyze its antibacterial activities, the maltose-sepharose (MS) beads specifically binding bovine SP-A was prepared by the covalently crosslinking method. The MS beads were used to absorb the bovine SP-A from bovine lung lavage. SDS-PAGE and Western blot detections showed that two forms of SP-A, 30 ku monomer and 60 ku dimer, were detected in the bovine SP-A preparation. The SP-A was identified as possessing the ability to aggregate E. coli, suggesting it possesses biological activity. The antibacterial experiment showed that SP-A significantly inhibited the growth of Staphylococcus aureus, Streptococcus agalactiae and the pathogenic E. coli, especially for Gram-negative bacteria with high inhibition activity, indicating that the bovine SP-A prepared in this study possesses antibacterial activities. This study provided necessary conditions for further investigation on SP-A biological characteristics and antibacterial activities, as well as opened up a new way for the development of SP-A antibacterial reagent.

Key words: bovine surfactant protein A, isolation and purification, meltose-sepharose beads, antibacterial activity

中图分类号:

S852.42

李楠, 张建楼, 张永红, 郑志强, 霍珊珊, 仲飞, 翟向和. 牛肺泡表面活性蛋白A的分离纯化及抗菌活性分析[J]. 畜牧兽医学报, 2021, 52(2): 498-505.

LI Nan, ZHANG Jianlou, ZHANG Yonghong, ZHENG Zhiqiang, HUO Shanshan, ZHONG Fei, ZHAI Xianghe. Preparation of Bovine Surfactant Protein A and Its Antibacterial Activities[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(2): 498-505.

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