| 金黄色葡萄球菌胞外分泌蛋白的核酸酶活性研究 |
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| 引用本文： | 李琦,廖成水,毛福超,王晓利,张梦珂,张晓洁,张春杰,李银聚,吴庭才,程相朝.金黄色葡萄球菌胞外分泌蛋白的核酸酶活性研究[J].中国预防兽医学报,2020(2):175-181. |
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| 作者姓名： | 李琦 廖成水 毛福超 王晓利 张梦珂 张晓洁 张春杰 李银聚 吴庭才 程相朝 |
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| 作者单位： | 河南科技大学动物科技学院/洛阳市活载体生物材料与动物疫病防控重点实验室;河南科技大学医学院;洛阳职业技术学院 |
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| 基金项目： | 河南科技大学博士启动基金项目(13480071);河南省科技攻关项目(182102110061);国家自然科学基金项目(31572489,31802159)。 |
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| 摘 要： | 为研究金黄色葡萄球菌胞外分泌蛋白的核酸酶活性,本研究复苏培养金黄色葡萄球菌后取培养上清液,采用透析得到金黄色葡萄球菌胞外分泌蛋白,结果显示获得的该蛋白浓度为48.5μg/mL。采用琼脂糖凝胶电泳法、琼脂扩散法和琼脂培养法检测金黄色葡萄球菌胞外分泌蛋白的核酸酶活性,利用琼脂糖凝胶电泳法探究温度、pH、金属离子对核酸酶活性的影响。结果显示,金黄色葡萄球菌胞外分泌蛋白表现出降解λDNA的核酸酶活性,且最适温度和pH值分别为50℃和9.0,在低温和酸性条件下核酸酶的活性较弱,但胞外核酸酶对70℃以上的耐受性较差。不同浓度的Ba^2+、Mg^2+和Zn^2+对胞外分泌蛋白的核酸酶活性无影响;低浓度(0.01 mmol/L^1 mmol/L)的Ca^2+、Ni^2+、Cu^2+和Mn^4+可以促进胞外核酸酶切割λDNA的活性;高浓度的Na^+、K^+和Fe^3+可以提高胞外核酸酶切割λDNA的活性;添加Co^2+(0.01 mmol/L^10 mmol/L)可以促进胞外分泌蛋白的核酸酶活性。本研究证实了金黄色葡萄球菌胞外分泌蛋白的核酸酶活性,为进一步研究胞外分泌蛋白在金黄色葡萄球菌和宿主互作中的确切作用奠定了基础。
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| 关 键 词： | 金黄色葡萄球菌 胞外蛋白 核酸酶 金属离子 活性 |
Characterization of the nuclease activity of extracellular proteins from Staphylococcus aureus |
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| LI Qi,LIAO Cheng-shui,MAO Fu-chao,WANG Xiao-li,ZHANG Meng-ke,ZHANG Xiao-jie,ZHANG Chun-jie,LI Yin-ju,WU Ting-cai,CHENG Xiang-chao.Characterization of the nuclease activity of extracellular proteins from Staphylococcus aureus[J].Chinese Journal of Preventive Veterinary Medicine,2020(2):175-181. |
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| Authors: | LI Qi LIAO Cheng-shui MAO Fu-chao WANG Xiao-li ZHANG Meng-ke ZHANG Xiao-jie ZHANG Chun-jie LI Yin-ju WU Ting-cai CHENG Xiang-chao |
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| Institution: | (College of Animal Science and Technology/Luoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control,Henan University of Science and Technology,Luoyang 471023,China;Medical College,Henan University of Science and Technology,Luoyang 471023,China;Luoyang Vocational and Technical College,Luoyang 471099,China) |
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| Abstract: | To investigate the nuclease activity of extracellular protein of Staphylococcus aureus(S.aureus).The extracellular protein was obtained from the supernatant fraction of S.aureus using dialysis bag and the concentration of purified protein is 48.5μg/mL.The nuclease activity of extracellular proteins of S.aureus was identified by agarose gel electrophoresis,agar diffusion method,and agar culture assay.Agarose gel electrophoresis was employed to determine the nuclease activity of extracellular proteins under different temperature,pH,and metal ions.The results showed that the extracellular proteins of S.aureus exhibited the nuclease activity againstλDNA,and the optimum temperature and pH were 50℃ and 9.0,respectively.The nuclease activity was weaker under a low temperature and acidic conditions,and the enzymes were inactivated when subjected to 70℃ or higher temperature Different concentrations of Ba^2+, Mg^2+, and Zn^2+ had no effect on the nuclease activity;however, the nuclease activity of extracellular proteins was increased at low ionic concentration(0.01 mmol/L-1 mmol/L) of Ca^2+, Ni^2+, Cu^2+, and Mn4+or at high ionic strength of Na^+, K^+, and Fe^3+, and the addition of Co^2+(0.01 mmol/L-10 mmol/L) promoted the degradation activity of λDNA. This study demonstrated that the extracellular proteins from S. aureus has nuclease activity, which lays a foundation for further study of the exact role of extracellular proteins in S. aureus and host interaction. |
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| Keywords: | Staphylococcus aureus extracellular protein nuclease metal ions activity |
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