文献类型：期刊文献

中文题名：中华眼镜蛇毒核糖核酸酶的分离纯化及其特殊生物学活性

英文题名：Special biological activities of a ribonuclease isolated from venom of Chinese cobra snake(Naja atra)

作者：陶凤云[1,2];赵伟[2];林强[2];马润宇[1]

第一作者：陶凤云

机构：[1]北京化工大学生命科学与技术学院;[2]北京联合大学生物化学工程学院

第一机构：北京化工大学生命科学与技术学院,北京100029

年份：2010

卷号：31

期号：3

起止页码：145-149

中文期刊名：中国生化药物杂志

外文期刊名：Chinese Journal of Biochemical Pharmaceutics

收录：北大核心:【北大核心2008】；CSCD:【CSCD_E2011_2012】；

基金：北京市教育委员会科技发展计划面上项目(KM200411417014)

语种：中文

中文关键词：中华眼镜蛇;核糖核酸酶;分离纯化;抑菌作用;细胞毒性;抗氧化

外文关键词：Chinese cobra snake; ribonuclease; isolation and purification; antibacterial effects; cytotoxic effects ; antioxidation

摘要：目的从中华眼镜蛇毒中分离纯化核糖核酸酶,并研究其生物学活性。方法以中华眼镜蛇毒为原料,采用SP-Trisacryl阳离子交换色谱、Sephadex G-75凝胶色谱、C8反相色谱等纯化方法,分离纯化具有核糖核酸酶活性的蛋白质,表征其酶学性质,检测其抑菌性、抗肿瘤细胞作用及抗氧化作用。结果 SDS-PAGE电泳显示纯化的中华眼镜蛇毒核糖核酸酶(Na-RNase)为相对分子质量为13 000的单一成分。该酶最适反应温度为40℃,最适pH值为6.5,米氏常数为3.67μmol/L,最大反应速率为3.52 pmol/s。在体外抑菌实验中,在8μmol/L的浓度下对大肠艾希菌和金黄葡萄球菌均未显示出显著抑制作用;在体外细胞毒性实验中,对于Hela肿瘤细胞和正常的人成纤维细胞在8μmol/L的浓度下无明显抑制作用;在抗氧化实验中,浓度达到71.5μmol/mL时,对小鼠肝微粒体脂质过氧化反应的抑制率为41.30%。结论中华眼镜蛇毒中的核糖核酸酶具有抗氧化作用,可能作为氧化还原的调节剂,在有关的疾病治疗方面具有应用的潜能。
Purpose A ribonuclease （Na-RNase） was purifed from venom of Chinese cobra snake （Naja atra） , and its biological activities were explored. Methods SP-Trisacryl cation-exchange chromatography, Sephadex G-75 gel chromatography and Cs reversed-phase chromatography were performed to obtain a purified Na-RNase. Its enzymatic properties were characterized, and antibacterial, antitumor and antioxidant effects were detected. Results SDS-PAGE electrophoresis of the purified Na-RNase shows a single band with relative molecular weight of 13 kDa. Its optimum temperature is 40 ℃ , and optimum pH is 6.5. Its Michaelis constant is 3.67 μmol/L, and maximum reaction rate is 3.52 pmol/s. In in vitro antibacterial tests for Escherichia coli and Staphylococcus aureus, no inhibitory effects were observed up to 8 μmol/L. And in in vitro cytotoxicity experiments for the Hela tumor cells and the normal human fibro-blasts cells, no cytotoxic effects were observed up to 8 μmol/L. In the antioxidation experiments, this Na-RNase was found to inhibit peroxidation of mice liver microsomal lipid, and the inhibitory rate is 41.30% at the Na-RNase concentration of 71.5 μmol/mL. Conclusion The Na-RNase isolated from Chinese cobra snake has antioxidant effect, and it may have application potential as a redox regulator in the treatment of diseases.