海洋微生物溶菌酶体外抗菌抗病毒活性研究

Study on Antimicrobial and Antiviral Activities in vitro of Marine Microorganism Lysozyme

【目的】初步研究海洋微生物溶菌酶S-12-86(MMLS)体外抑菌与抗病毒活性。【方法】采用最小抑菌浓度(MIC)和最小杀菌浓度法(MBC)测定MMLS对部分模式菌的抑菌和杀菌作用;通过细胞病变抑制实验法(CPE),研究MMLS在猪肾细胞(PK-15)中对伪狂犬病毒(PRV)的抑制作用。【结果】MMLS对革兰氏阳性菌,革兰氏阴性菌及真菌均有抑菌作用,抑菌作用的浓度范围在0.25～4.00mg·ml-1之间,杀菌作用的浓度范围一般在0.25～8.00mg·ml-1之间。在电镜下,可见MMLS能够使大部分白色念珠球菌的细胞壁出现严重变形,细胞质出现不均匀,细菌胞质中的空腔增多。MMLS的TC50为100.0μg·ml-1,抑制伪狂犬病毒的半数有效浓度(EC50)为0.46μg·ml-1,治疗指数(TI)为217;在病毒感染后的0、2、4、6和8h添加MMLS,均有抑制伪狂犬病毒的作用。【结论】MMLS具有广谱的抑菌作用和抗伪狂犬病毒作用。

[Objective] The in vitro antimicrobial and antiviral activities of marine microorganism lysozyme S-12-86 （MMLS） were studied. [Method] The minimum inhibition concentration （MIC） and the minimum bactericidal concentration （MBC） were used to test the antimicrobial effects of MMLS. The inhibiting effects of MMLS on Pseudo rabies Virus （PRV） in swine kidney cells （PK-15 cells） were judged by cytopathogenic effect test （CPE）. [ Result ] MMLS had a broad antimicrobial spectrum against several standard strains including G^＋, G^-and fungi, et al. The minimum inhibition concentration （MIC） was 0.25-4.00 mg.ml^-1 and the minimum bactericidal concentration （MBC） was 0.25-8.00 mg.ml^-1, respectively. Observation under the transmission electron microscope revealed that the cell wall of Candida albicans was seriously distortional, and the cytoplasm was asymmetrical. There were many cavities in the cytoplasm. The median toxic concentration （TC50） of MMLS was 100.0 μg.ml^-1, the median effective concentration （EC50） of MMLS was 0.46 μg.ml^-1, the selectivity index was 217. In time of addition experiment, MMLS inhibited PRV in PK-15 cell when it was added at 0, 2, 4, 6, and 8 h after virus infection. [Conclusion] We could conclude that MMLS had broad antimicrobial spectrum and good inhibiting effects on PRV.