罗非鱼肌肉乙酰胆碱酯酶的制备及冻干稳定性研究

Preparation of Muscle Acetylcholinesterase in Oreochromis aurea and Its Stability Analysis of Freeze-drying

以罗非鱼（Oreochromis aurea）肌肉组织为材料提取乙酰胆碱酯酶,采用正交试验法研究了不同提取条件（离心转速、提取液质量体积比W/V、匀浆时间）下肌肉粗酶液的酶活力,得到提取酶的最佳条件组合为离心转速6000rmp,提取液质量体积比W/V为1：4,匀浆时间为20s。在酶的冷冻干燥稳定性实验中,添加5%聚乙二醇或5%蔗糖作为稳定剂能有效保护酶活力,冻干时剩余酶活力约为80%。作为保护剂5%聚乙二醇和5%蔗糖两者的差异不显著。酶干粉保存15d时,三种保存温度下（-20℃,0-4℃,20-25℃）,添加保护剂与不添加保护剂的酶活力差值约为10%;30d时,差值为10%-15%;60d时,差值为30%~40%。随着保存温度的提高,添加保护剂的稳定效应逐渐被较高温度对酶活力的不利影响抵消。添加5%聚乙二醇作为稳定剂,-20℃,60d时,酶干粉测得剩余酶活力为74%。

This paper is aimed to present the author＇s research on preparation and freeze-drying stability of muscle acetylcholinester-ase （ACHE） in Oreochromis aurea. Using orthogonal test, the optimal extraction condition was： 6000rpm, w/v of extraction as 1：4, ho-mogenizing time as 20s. In research of freeze-drying stability, 5% polyethylene glycol or 5% sucrose are effective in protecting enzymeactivity, with remaining enzyme activity as 80%. There is no significant difference between 5% polyethylene glycol and 5% sucrose. Af-ter 15 days, with or without protective agent, the difference of enzyme activity among three temperature （- 20 ℃ , 0-4 ℃ , 20- 25 ℃ ） is10%, and after 30 days, the difference is 10%-15%, and after 60 days, the difference is 30%-40% , which means as temperature ris-es, the effect of protective agent would be neutralized. With 5% polyethylene glycol as protective agent, the remaining enzyme activityis 74% 60 days under -20℃ after freeze-drying.