Abstract
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.
Highlights
Superoxide Dismutase (SOD) plays a central role in the antioxidant system [1] because of its specific catalysis of the dismutation of the primary free radical—superoxide anion [2], which endows it with attractive potential in the fields of medicine and functional food
The Maillard reaction of the co-incubated reactant of Cu/Zn SOD and glucose was investigated by SDS-PAGE
The un-incubated Cu/Zn SOD exhibited a main protein band with a molecular weight of about 16 KDa, while in the case of G-SOD, the density of this band decreased with an obvious tailing phenomenon, and a protein band with a molecular weight higher than 116 KDa appeared with obvious tailing. These results indicated that the Maillard reaction occurred during the co-incubation process, and that some polymer aggregate of Cu/Zn SOD developed in the reactant [12]
Summary
Superoxide Dismutase (SOD) plays a central role in the antioxidant system [1] because of its specific catalysis of the dismutation of the primary free radical—superoxide anion [2], which endows it with attractive potential in the fields of medicine and functional food. A few preparative methods, for example polyketal microparticles, provide relatively safe and efficient delivery for SOD [8,9], most of these previous preparative procedures require either complicated operations, a variety of reagents—some of them even highly toxic—and low encapsulation efficiency or low release rate. These disadvantages have limited the applications of SOD NPs. more alternative simple and green preparation methods are desirable
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