摘要
以克氏原螯虾(Procambarus clarkii)酚氧化酶(PO)为材料,采用L-DOPA为反应底物,分光光度计测定不同温度和pH条件下PO活力,研究克氏原螯虾PO的特性,为其免疫学研究及优化PO准确测定方法提供依据。结果显示PO活力最高时的温度为35℃,低于和高于35℃活力逐渐下降,45℃以上活力迅速下降;0℃仍具有最高活力的24%。在pH4.0~9.5内PO均具有活力,鳃组织最佳pH5.5,血清最佳pH6.5。因此测定克氏原螯虾PO活力的最佳参数为温度35℃,pH5.5(鳃)或6.5(血清)。以上结果说明冰浴终止反应法测定克氏原螯虾PO活力会影响结果的准确性,要慎用。
The characteristic of phenoloxidase in the red swamp crayfish ( Procambarus clarkii) was studied for immunology research and accurate determination method in this experiment. The phenoloxidase activity of gills and serum in different temperature and pH was determined with spectrophotometer based on L-DOPA as substrate. The result showed that the optimum temperature of phenoloxidase was 35 ℃. The activity of phenoloxidase decreased gradually while the temperature was lower than 35 ℃ or more than 35 ℃, and dropped promptly when the temperature rose above 45 ℃. The activity of phenoloxidase in 0 ℃ was 24% of the highest in 35 ℃. Phenoloxidase had activity when pH was 4 -9. 5. The most suitable pH was 5.5 in gills and 6. 5 in serum. For this reason, the best parameter of phenoloxidase activity determining in Procambarus clarkii was 35 ℃ and pH 5.5 (gills) or 6. 5 (serum). At the same time icebath used in determining to terminate the reaction of L-DOPA and phenoloxidase was not good.
出处
《淡水渔业》
CSCD
北大核心
2012年第6期68-72,共5页
Freshwater Fisheries
基金
江苏牧院科技助农项目(ZN1208)
江苏省水产三项工程项目(PJ2011-56)