葡萄糖氧化酶的提纯方法研究

Study on the method of purification of GOD

主要研究葡萄糖氧化酶从废菌丝中的提取方法。由于葡萄糖氧化酶是胞内酶,需要先将菌丝破壁,使其中的葡萄糖氧化酶释放出来,而且要采取措施保证酶的活力损失最小。获得的初酶液,再经过初步纯化,即可制成酶制剂。最终确定了破壁的方法为超声破壁,超声破壁的最优条件为:超声时间50min,超声温度60℃,超声功率200W。破壁后选取65%乙醇一次沉淀葡萄糖氧化酶,然后用16%的PEG二次沉淀酶。

This paper mainly consider the method of purification of Glucose Oxidase from waste mycelia. Because GOD is intraeellular enzyme, first mycelium need to be broken, then glucose oxidase can be released from intracellular, at the same time I should take steps to ensure minimal loss of enzyme activity. After the initial enzyme solution is obtained, it is luther to be purifieated and be made into enzymes. Fi- nally the method of waste mycelia broken can be determined, the optimal conditions are： ultrasonic time is 50 min, ultrasound temperatures is 60 %, the ultrasonic power is 200 W. After waste mycelia is bro- ken,65% ethanol is used to precipite GOD, and then 16% PEG is used to precipite GOD for the second time.