金、银纳米颗粒固定化重组人精氨酸酶I:制备及性能研究

N. Stasyuk, R. Serkiz, S. Mudry, G. Gayda, Andriy Zakalskiy, Yevgen Koval’chuk, M. Gonchar, M. Nisnevitch
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引用次数: 16

摘要

金属纳米粒子(NPs),如金(Au)和银(Ag),由于其独特的光学、电学和光热性质,在化学、物理和生物学中具有重要意义。这些NPs被广泛用于固定各种生物活性物质,包括肽、酶、抗体和DNA。分别用葡萄糖还原硝酸银和柠檬酸钠还原四氯金酸制备了纳米银和纳米金。采用TEM、AFM和XRD等方法对AgNPs和AuNPs的尺寸和结构进行了表征。AgNPs和AuNPs的平均大小在8 ~ 15 nm之间。用碳二亚胺五氟苯酚法将重组精氨酸酶I固定在ω-巯基十六烷酸功能化的NPs表面。结果表明,从酵母中分离的重组人肝精氨酸酶I在两种NPs上固定后都保持了良好的稳定性。与游离精氨酸酶相比,在+4℃条件下保存25天后,固定化后的精氨酸酶在AuNPs表面和AgNPs表面的活性分别保持了40%和25%。该固定化酶可用于药品、食品和血液中精氨酸的测定。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Recombinant human arginase I immobilized on gold and silver nanoparticles: preparation and properties
Metal nanoparticles (NPs), such as gold (Au) and silver (Ag), are important for chemistry, physics, and biology due to their unique optical, electrical, and photothermal properties. Such NPs are widely used for immobilization of various bioactive substances, including peptides, enzymes, antibodies and DNA. The synthesis of silver and gold nanoparticles was carried out by reduction of silver nitrate by glucose and reduction of tetrachloroauric acid by sodium citrate, respectively. The size and structure of the AgNPs and AuNPs were characterized using TEM, AFM and XRD methods. The average size of the AgNPs and AuNPs was between 8 and 15 nm. Recombinant arginase I was immobilized using the carbodiimidepentafluorophenol method on the surface of NPs functionalized with ω-mercaptohexadecanoic acid. It was shown that recombinant human liver arginase I isolated from the yeast Hansenula polymorpha maintains satisfactory stability after immobilization on both NPs. The immobilized arginase retained 40% of its activity on the surface of AuNPs and 25% on AgNPs compared to the free arginase after storage at +4 oC during 25 days. The immobilized enzyme can be used for assay of arginine in pharmaceuticals, in food products and in blood.
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