School of Food Science and Technology, Jiangnan University, Wuxi, China.
J Sci Food Agric. 2012 Mar 15;92(4):787-93. doi: 10.1002/jsfa.4646. Epub 2011 Nov 18.
High-purity soybean phosphatidylcholine (SPC) (94%) were prepared using macroporous resin adsorption chromatography previously. Catalase is a food enzyme for promoting health and protecting against many age-related disease. Solid lipid nanoparticles (SLN) are safe immobilizing systems for efficient protein transportation to biomembranes while avoiding adverse degradation of protein. This study was aimed at developing and characterizing catalase-loaded SLN using SPC as solubilizers and stabilizing agents, to protect catalase from proteolysis.
Catalase-loaded SLN were prepared by the double emulsification method and solvent evaporation techniques, using acetone-methylene chloride (1:1, v/v) as an organic solvent, SPC-tripalmitin as oil phase and Poloxamer 188 as a surfactant. The optimized SLN were prepared using an SPC:tripalmitin ratio of 5% (w/w), 20 s plus 30 s sonication, 20 g L⁻¹ Poloxamer 188 and 1:2 (v/v) of oily phase:outer aqueous phase ratio. The mean particle size of SLN was 296.0 ± 7.0 nm, polydispersity index range and zeta potential were 0.322-0.354 and -36.4 ± 0.6, respectively, and encapsulation efficiency reached its maximum of 77.9 ± 1.56%. Catalase, which was found to distribute between the solid lipid and inner aqueous phase, was gradually released from SLN up to 20% within 20 h. Catalase-loaded SLN had stably retained 30% of H₂O₂-degrading activity for at least 24 h in a proteolytic environment, while free catalase lost its activity within 1 h.
Catalase can indeed be loaded in tripalmitin-based SLN using SPC as solubilizers and stabilizing agents, which protected it against proteolysis, suggesting the potential application of SPC in delivery and protection of functional food enzyme.
高纯度大豆卵磷脂（SPC）（94%）以前是通过大孔树脂吸附色谱法制备的。过氧化氢酶是一种促进健康和预防许多与年龄相关疾病的食品酶。固体脂质纳米粒（SLN）是一种安全的固定化系统，可有效将蛋白质输送到生物膜，同时避免蛋白质的不良降解。本研究旨在开发并表征负载过氧化氢酶的 SLN，使用 SPC 作为增溶剂和稳定剂，以保护过氧化氢酶免受蛋白水解。
采用双重乳化法和溶剂蒸发技术制备负载过氧化氢酶的 SLN，使用丙酮-二氯甲烷（1:1，v/v）作为有机溶剂，SPC-三棕榈酸甘油酯作为油相，泊洛沙姆 188 作为表面活性剂。使用 SPC:三棕榈酸甘油酯比例为 5%（w/w）、20 s 加 30 s 超声、20 g/L 泊洛沙姆 188 和油相：外水相比例为 1:2（v/v）优化 SLN。SLN 的平均粒径为 296.0±7.0nm，多分散指数范围和 Zeta 电位分别为 0.322-0.354 和-36.4±0.6，包封效率最高可达 77.9±1.56%。发现过氧化氢酶分布在固体脂质和内水相之间，在 20 h 内从 SLN 中逐渐释放，达到 20%。在蛋白水解环境中，负载过氧化氢酶的 SLN 稳定保留至少 24 h 的 30%H₂O₂降解活性，而游离过氧化氢酶在 1 h 内失去活性。
确实可以使用 SPC 作为增溶剂和稳定剂将过氧化氢酶负载在基于三棕榈酸甘油酯的 SLN 中，从而保护其免受蛋白水解，这表明 SPC 在功能性食品酶的递送和保护方面具有潜在应用。
