TTC法测定离子液体中产紫青霉细胞存活率的研究

Cell Survival Determination of Penicillium purpurogenum Li-3 by TTC Assay in Different Ionic Liquids

以产紫青霉菌细胞为供试材料,采用2,3,5-氯化三苯基四氮唑(TTC)测定了不同波长、萃取剂、显色时间、显色温度以及不同pH下的脱氢酶活性,并将优化后的TTC-脱氢酶还原法用于测定不同离子液体中产紫青霉的细胞存活率。结果表明,在0.5%的TTC缓冲溶液中,测定波长为485 nm时吸光度最高;选用丙酮作为萃取剂时,萃取效果好;适宜的显色时间为1 h,显色温度为32℃;在pH为7.0、0.2 mol/L的NaH2PO4-Na2HPO4(PB)缓冲溶液中,显色稳定,脱氢酶活性也较好。优化后的TTC-脱氢酶还原法能较为准确、快速的检测出不同离子液体中产紫青霉菌的细胞存活率。

Using Penicillium purpurogenum Li-3 cells as experimental material, dehydrogenase activity was measured by 2,3,5- three triphenyl tetrazolium chloride （TTC） dehydrogenase reduction method at different wavelengths, extraction agent, chromogenic time, coloration temperature and pH values, and the optimized TYC-dehydrogenase reduction method was chosen for determination of survival rate of Penicillium purpurogenum Li-3 in different ionic liquids. The results showed that the absorbance value reached the highest in 0.5% TTC buffer solution under 485 nm detection wavelength; good extraction effect was achieved using acetone as extraction agent; optimum chromogenic time is 1.5 h at the temperature of 32C; The dehydrogenase activity was stable in pH 7.0 0.2 mol/L NaH2PO4-NazHPO4 （PB） buffer solution. The optimized TrC- dehydrogenase reduction method could detect the survival rate ofPenicilliurn purpurogenum Li-3 in different ionic liquids accurately and rapidly.