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Preparation and Characterization of Nanoparticles Made from Co-Incubation of SOD and Glucose

机译：SOD与葡萄糖共温育纳米粒子的制备与表征

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摘要

The attractive potential of natural superoxide dismutase (SOD) in the fields of medicine and functional food is limited by its short half-life in circulation and poor permeability across the cell membrane. The nanoparticle form of SOD might overcome these limitations. However, most preparative methods have disadvantages, such as complicated operation, a variety of reagents—some of them even highly toxic—and low encapsulation efficiency or low release rate. The aim of this study is to present a simple and green approach for the preparation of SOD nanoparticles (NPs) by means of co-incubation of Cu/Zn SOD with glucose. This method was designed to prepare nanoscale aggregates based on the possible inhibitory effect of Maillard reaction on heating-induced aggregation during the co-incubation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results indicated that the Maillard reaction occurred during the co-incubation process. It was found that enzymatically active NPs of Cu/Zn SOD were simultaneously generated during the reaction, with an average particle size of 175.86 ± 0.71 nm, and a Zeta potential of −17.27 ± 0.59 mV, as established by the measurement of enzymatic activity, observations using field emission scanning electron microscope, and analysis of dynamic light scattering, respectively. The preparative conditions for the SOD NPs were optimized by response surface design to increase SOD activity 20.43 fold. These SOD NPs showed storage stability for 25 days and better cell uptake efficacy than natural SOD. Therefore, these NPs of SOD are expected to be a potential drug candidate or functional food factor. To our knowledge, this is the first report on the preparation of nanoparticles possessing the bioactivity of the graft component protein, using the simple and green approach of co-incubation with glucose, which occurs frequently in the food industry during thermal processing.
机译：天然超氧化物歧化酶（SOD）在医药和功能食品领域的诱人潜力受到其半衰期短，细胞膜通透性差的限制。 SOD的纳米颗粒形式可能会克服这些限制。但是，大多数制备方法都具有缺点，例如操作复杂，试剂种类繁多（其中有些甚至具有剧毒），包封效率低或释放速率低。这项研究的目的是提出一种简单而绿色的方法，通过将Cu / Zn SOD与葡萄糖共孵育来制备SOD纳米颗粒（NP）。根据美拉德反应在共孵育过程中对加热诱导的聚集的可能抑制作用，设计该方法来制备纳米级聚集体。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）结果表明，在共孵育过程中发生了美拉德反应。发现在反应过程中同时生成了具有酶活性的Cu / Zn SOD NP，其平均粒径为175.86±0.71 nm，Zeta电位为-17.27±0.59 mV，这是通过测定酶活性确定的，使用场发射扫描电子显微镜进行观测，并分别进行动态光散射分析。通过响应面设计优化了SOD NP的制备条件，以使SOD活性增加20.43倍。与天然SOD相比，这些SOD NP表现出25天的储存稳定性和更好的细胞吸收功效。因此，这些SOD NP有望成为潜在的候选药物或功能性食品因子。据我们所知，这是有关使用具有葡萄糖共孵育作用的简单绿色方法制备具有移植物成分蛋白生物活性的纳米颗粒的首次报道，这种方法在食品加工过程中经常发生。

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