华南农业大学学报2017，Vol.38Issue(5)：19-23,5.DOI:10.7671/j.issn.1001-411X.2017.05.004
A型产气荚膜梭菌噬菌体裂解酶Cp51的原核表达及活性检测
Prokaryotic expression and activity detection of bacteriophage lysin Cp51 against Clostridium perfringens type A
摘要
Abstract
[Objective] To construct a prokaryotic expression system of type A Clostridiumperfringens phage lysin Cp.51,and study its antibacterial activity against C Perfringens type A in vitro.[Method] Bacteriophage lysin Cp51 gene was synthesized.The prokaryotic expression vector pET-32a-Cp51 was constructed and transformed into Escherichia coli BL21 (DE3).After induction using 0.5 mmol·L-1 IPTG,the soluble recombinant protein Cp51 was successfully expressed,and was subsequently purified with Ni2+-NTA affinity chromatography.The antibacterial activity of the recombinant protein Cp51 was detected by kinetic turbidimetric assay.[Result] The bacteria turbidities of seven strains of C.perfringens type A were effectively reduced by the recombinant protein Cp51.The bactericidal rate was above 99.99％ in 30 min after treatment of recombinant protein Cp51 at the concentration of above 5 μg·mL-1.The Cp51 protein had no bactericidal effect against other types of bacteria.[Conclusion] The recombinant protein of bacteriophage lysine Cp51 has strong bactericidal activity and specificity in vitro against type A C.perfringens,which could provide a basis for clinical application of Cp51 lysin.关键词
产气荚膜梭菌/噬菌体/裂解酶/蛋白表达/杀菌活性Key words
Clostridium perfringens/bacteriophage/lysin/protein expression/bactericidal activity分类
生物科学引用本文复制引用
吕梦娜,龙航宇,王丽梅,唐陶,陈冰冰,林瑞庆..A型产气荚膜梭菌噬菌体裂解酶Cp51的原核表达及活性检测[J].华南农业大学学报,2017,38(5):19-23,5.基金项目
广东省科技计划项目(2014A020208099) （2014A020208099）
