一株拮抗绿脓杆菌的抗菌蛋白产生菌的分离与鉴定

Isolation and Identification of an Antimicrobial Protein-producing Strain Against Pseudomonas Aeruginosa

获得了产生拮抗绿脓杆菌抗菌蛋白的菌株。从土壤样品中筛选拮抗绿脓杆菌的菌株,采用形态学和16S rDNA分析鉴定其分类地位,采用琼脂平板扩散法测定分离菌发酵产物拮抗绿脓杆菌的活性,采用硫酸铵沉淀、分子筛分离发酵上清中的目标蛋白,测定了目标蛋白的蛋白酶敏感性、高温耐受性。结果表明:从土壤中分离出的该株不动杆菌,从其发酵产物中纯化出的目标蛋白有拮抗绿脓杆菌的活性;该目标蛋白对蛋白酶K或高温敏感,处理后可降低或丧失抗绿脓杆菌的活性,该抗菌蛋白相对分子量约35 KD。

To screen and indentify a strain expression antimicrobial protein against Pseudomonas aeruginosa,the strain was isolated from soil samples and then identified according to the morphological features and 16S rDNA sequence analysis.Agar diffusion method was performed to determine the antimicrobial activity of target protein produced from the isolated strain.Ammonium sulfate precipitation and molecular sieve were used to purify antibacterial protein from fermentation liquor.Protease K sensitivity and high temperature tolerance were detected to analyze the stability of antibacterial protein.A strain,expression antimicrobial protein against Pseudomonas aeruginosa was screened out,which belonged to Acinetobacter.The target protein was purified from fermentation liquor,but its antimicrobial activity was instable.Protease K or high temperature treatment would cause target proteint's antimicrobial activity reduced even lost.The molecular weight of antimicrobial protein was 35 KD.