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目的研究Bicine-Na OH缓冲液制备的尿酸酶(UCA)-过氧化氢酶纳米脂质体(UCNL)中UCA的生化特性及初步稳定性。方法使用Bicine-Na OH缓冲液并采用逆向蒸发法制备UCNL,考察UCNL的形态结构、粒径分布、Zeta电位、包封率及UCNL中UCA的最适温度、最适p H、米氏常数和热稳定性、贮存稳定性、酸碱稳定性、抗胰蛋白酶水解能力和抗部分金属离子和有机化合物的能力。结果 UCNL呈类圆形,包封率为56.87%±3.81%,粒径为759.30±28.12 nm,Zeta电位为-8.49±3.18 m V。UCNL中UCA和游离UCA的最适温度均为40℃,最适p H分别为8.0、8.5,Km值分别13.40、13.87μmol·L-1;UCNL中UCA的各种稳定性均比游离UCA好。结论 UCNL中UCA的催化活性和稳定性较游离酶明显增加。
Objective To study the biochemical characteristics and preliminary stability of UCA in uricase (UCA) - catalase nano-liposome (UCNL) prepared by Bicine-Na OH buffer. Methods UCNL was prepared by Bicine-Na OH buffer and reverse-evaporation method. The morphological structure, particle size distribution, Zeta potential and entrapment efficiency of UCNL were investigated. The optimum temperature of UCA in UCNL, Thermal stability, storage stability, acid-base stability, antitrypsin hydrolytic ability and resistance to partial metal ions and organic compounds. Results The UCNLs were round in shape with entrapment efficiency of 56.87% ± 3.81%, particle size of 759.30 ± 28.12 nm and Zeta potential of -8.49 ± 3.18 mV. The optimum temperature of UCA and free UCA in UCNL were both 40 ℃, the optimum p H were 8.0, 8.5 and the Km values were 13.40 and 13.87μmol·L-1, respectively. The stability of UCA in UCNL was better than that of free UCA . Conclusion The catalytic activity and stability of UCA in UCNL are significantly higher than that of free enzyme.