HIGHLY STABLE AND RECYCLABLE ENZYME-CALCIUM PHOSPHATE NANOPARTICLES

摘要

PURPOSE: A nanoparticle with a complex is provided to ensure high enzyme loading ability and stability. CONSTITUTION: A nanoparticle contains a complex by fixation of enzyme to calcium phosphate. The nanoparticle further contains sodium citrate. The enzyme is esterase, protease, glucose oxidase, or alpha-chymotrypsin. The size of the nanoparticles is 200-300 nm. The nanoparticle has stability and reusability. A method for preparing CP-enzyme nanoparticles comprises: a step of putting 2.65 ml of cylcohexane and 30 vol% of lgepal CO-520 into each 10ml of glass bottle; a step of adding 100 ul of 60mM NaHPO_4(B) containing 20-100 ul of various esterase enzymes(10-50 mg/ml) and 120 ul of 100 mM CaCl_2(A) containing water soluble phase to the each glass bottle; a step of stirring at 600 rpm for 20 minutes to obtain transparent micromeulsIon A and B; a step of dropping enzyme microemulsion to the microemulsion A; a step of completely mixing both emulsions and stirring for 5 minutes; a step of stabilizing calcium phosphate nanoparticles; a step of adding 50 ul of sodium citrate(15mM) and reacting for 20 minutes; a step of adding three times volume of ethanol and pulverizing the emulsion; a step of adding ethanol and preparing calcium phosphate particles of transparent fluffy precipitation; and a step of centrifuging the solution at 6000 rpm; and a step of washing with ethanol 3-4 times and vacuum-drying.