Chemistry Department, Missouri State University, 901 S. National Ave, Springfield, Missouri 65897, USA.
Biomater Sci. 2019 May 28;7(6):2511-2519. doi: 10.1039/c9bm00129h.
Herein, we demonstrate the use of lysozyme (Lys) as a model to fabricate a protein carrier system based on gold nanoparticles (AuNPs) via the Layer-by-Layer (LbL) technology. Poly(ethyleneimine) (PEI) and poly(sodium 4-styrenesulfonate) (PSS) were used as cationic and anionic polymers respectively to grow oppositely charged layers. Mild aqueous conditions were utilized to avoid protein denaturation and activity instead of organic solvents that have been used in other encapsulation systems. Two different strategies were used: (A) lysozyme acting as a reducing and stabilizing agent in the formation of AuNPs at a temperature of 45 ± 2 °C followed by only two subsequent polymeric layers deposited by LbL, and (B) citrate acting as a reducing agent prior to stabilization of the AuNPs by mercaptoundecanoic acid. Dynamic light scattering, UV-vis spectroscopy, IR spectroscopy and transmission electron microscopy were used to characterize the nanoconjugates. Furthermore, the enzymatic activity of the resulting protein/nanoparticle conjugates was evaluated using the bacteria Micrococcus lysodeikticus as a substrate.
在这里，我们展示了使用溶菌酶 (Lys) 作为模型，通过层层 (LbL) 技术制备基于金纳米粒子 (AuNPs) 的蛋白质载体系统。聚（聚乙烯亚胺）(PEI) 和聚（4-苯乙烯磺酸钠）(PSS) 分别用作阳离子和阴离子聚合物，以生长带相反电荷的层。采用温和的水相条件避免了蛋白质变性和失活，而不是其他封装系统中使用的有机溶剂。使用了两种不同的策略：(A) 在 45±2°C 的温度下，溶菌酶作为 AuNPs 形成的还原剂和稳定剂，随后仅通过 LbL 沉积两个后续的聚合物层，和 (B) 在巯基十一酸稳定 AuNPs 之前，柠檬酸作为还原剂。动态光散射、紫外可见光谱、红外光谱和透射电子显微镜用于表征纳米复合物。此外，使用细菌 Micrococcus lysodeikticus 作为底物评估了所得蛋白质/纳米粒子缀合物的酶活性。
