摘要
对生物战剂及病原微生物实现快速、安全、高效的大面积消除或抑制是生物安全领域消毒技术研究的重点。本研究以碳纳米管为固定化载体,采用EDC-NHS共价交联法和浸渍法负载溶菌酶和纳米银制备新型抑菌复合材料CNTs-Lzm-AgNps复合纳米酶,考察了溶菌酶浓度、缓冲液pH、固定温度、固定时间等条件对溶菌酶固定情况的影响。结果表明,当初始溶菌酶浓度3.0 mg/mL、缓冲液pH=10.0、固定温度25℃,固定时间12 h时,溶菌酶的固定效果最佳。此时溶菌酶负载量可达280.2 mg/g,相对酶活性为88.1%。当硝酸银与溶菌酶摩尔比为25∶1时,所制备的材料具有合适的纳米银负载量且不会影响酶活性。此时材料中的银含量为3.41%±0.07%,银的负载率为63.26%±1.28%,相对酶活性为86.6%。用傅里叶红外光谱仪(FTIR)、扫描电子显微镜(SEM)、透射电子显微镜(TEM)、能量色散X射线光谱仪(EDX)和X射线衍射仪(XRD)对样品进行表征,表明该复合纳米酶负载均匀,填充效果好。对该复合纳米酶的抑菌效果进行研究,发现其在温度30℃、pH=5.0时对大肠杆菌和金黄色葡萄球菌的抑菌效果最佳,证明该复合纳米酶是一种稳定、广谱、绿色的抑菌剂,具有实际应用的可行性。
Rapid,safe and efficient elimination or inhibition of biological warfare agents and pathogenic microorganisms is the focus of disinfection technology research in the field of biosafety.In this study,CNTs-Lzm-AgNps composite nano-enzyme was prepared as a new antibacterial composite by using carbon nanotubes as immobilization carrier,loading lysozyme and nano-silver with EDC-NHS covalent crosslinking method and impregnation method.The effects of lysozyme concentration,buffer pH,fixing temperature and fixing time on the immobilization of lysozyme were investigated.The results showed that when the initial lysozyme concentration was 3.0 mg/mL,the buffer pH was 10.0,the fixing temperature was 25℃,and the fixing time was 12 hours,the immobilization effect of lysozyme was the best.Under these conditions,the load of lysozyme reached 280.2 mg/g,and the relative enzyme activity was 88.1%.When the molar ratio of silver nitrate to lysozyme was 25:1,the prepared material has appropriate silver loading capacity without affecting the enzyme activity.At this time,the silver content in the material was 3.41%±0.07%,the silver loading rate was 63.26%±1.28%,and the relative enzyme activity was 86.6%.The samples were characterized by Fourier transform infrared spectroscopy(FTIR),scanning electron microscopy(SEM),transmission electron microscopy(TEM),energy dispersive X-ray spectroscopy(EDX)and X-ray diffraction(XRD).The results showed that the composite nano-enzyme was uniformly loaded and had a good filling effect.The bacteriostatic efficacy of the composite nano-enzyme was studied.It is found that the bacteriostatic efficacy of the composite nano-enzyme on Escherichia coli and Staphylococcus aureus is the best at 30℃and pH=5.0,which proves that the composite nano-enzyme is a stable,broad-spectrum and green bacteriostatic agent with practical application feasibility.
作者
马腾博
陈高云
赵传欣
卢穆青
刘敏
MA Tengbo;CHEN Gaoyun;ZHAO Chuanxin;LU Muqing;LIU Min(Army Institute of NBC Defence,Beijing 102205,China)
出处
《防化研究》
2023年第1期46-55,共10页
CBRN DEFENSE
关键词
碳纳米管
溶菌酶
纳米银
固定化
复合抑菌材料
抑菌效果
复合纳米酶
carbon nanotube
lysozyme
nano silver
immobilization
composite antibacterial material
bacteriostasis efficacy
composite nano-enzyme
