Objective: The integrated method was investigated to measure Vm/Km of mouse liver glutathione S-transfer-ase (GST) activity on GSH and 7-Cl-4-nitrobenzofurazozan. Methods: Presetting concentration of one substrate twe...Objective: The integrated method was investigated to measure Vm/Km of mouse liver glutathione S-transfer-ase (GST) activity on GSH and 7-Cl-4-nitrobenzofurazozan. Methods: Presetting concentration of one substrate twenty-fold above the other's and taking maximum product absorbance Am as parameter while Km as constant, Vm/Km was obtained by nonlinear fitting of GST reaction curve to the integrated Michaelis-Menten equation In [Am/(Am -Ai)] + Ai/ ( ξ× Km ) = ( Vm/Km )×ti (1). Results: Vm/Km for GST showed slight dependence on initial substrate concentration and data range, but it was resistant to background absorbance, error in reaction origin and small deviation in presetting Km. Vm/Km was proportional to the amount of GST with upper limit higher than that by initial rate. There was close correlation between Vm/Km and initial rate of the same GST. Consistent results were obtained by this integrated method and classical initial rate method for the measurement of mouse liver GST. Conclusion: With the concentration of one substrate twenty-fold above the other's, this integrated method was reliable to measure the activity of enzyme on two substrates , and substrate concentration of the lower one close to its apparent Km was able to be used.展开更多
Excessive nitrate in groundwater has emerged as a serious environmental concern. The elevated nitrate concentration in drinking water causes a serious threat to public health on account of the possible transformation ...Excessive nitrate in groundwater has emerged as a serious environmental concern. The elevated nitrate concentration in drinking water causes a serious threat to public health on account of the possible transformation of nitrate to nitrite, which is one of the main predisposing factors of methemoglobinemia [1].展开更多
本文探讨了超声处理对戊二醛改性明胶包埋的木瓜蛋白酶酶解反应条件以及酶学特性的影响。采用均匀实验的方法优化超声处理固定化木瓜蛋白酶的反应条件。利用偏最小二乘回归法分析均匀实验结果,得到最优条件为:135 k Hz、0.05 W/cm2、20 ...本文探讨了超声处理对戊二醛改性明胶包埋的木瓜蛋白酶酶解反应条件以及酶学特性的影响。采用均匀实验的方法优化超声处理固定化木瓜蛋白酶的反应条件。利用偏最小二乘回归法分析均匀实验结果,得到最优条件为:135 k Hz、0.05 W/cm2、20 min、40℃,此条件下实测相对酶活为221.25%,与理论值相对误差为3.58%。超声并没有改变固定化酶的最适p H和最适温度,即超声并不改变固定化酶的酶学特性。展开更多
基金National Natural Science Foundation of China (No.30200266)
文摘Objective: The integrated method was investigated to measure Vm/Km of mouse liver glutathione S-transfer-ase (GST) activity on GSH and 7-Cl-4-nitrobenzofurazozan. Methods: Presetting concentration of one substrate twenty-fold above the other's and taking maximum product absorbance Am as parameter while Km as constant, Vm/Km was obtained by nonlinear fitting of GST reaction curve to the integrated Michaelis-Menten equation In [Am/(Am -Ai)] + Ai/ ( ξ× Km ) = ( Vm/Km )×ti (1). Results: Vm/Km for GST showed slight dependence on initial substrate concentration and data range, but it was resistant to background absorbance, error in reaction origin and small deviation in presetting Km. Vm/Km was proportional to the amount of GST with upper limit higher than that by initial rate. There was close correlation between Vm/Km and initial rate of the same GST. Consistent results were obtained by this integrated method and classical initial rate method for the measurement of mouse liver GST. Conclusion: With the concentration of one substrate twenty-fold above the other's, this integrated method was reliable to measure the activity of enzyme on two substrates , and substrate concentration of the lower one close to its apparent Km was able to be used.
文摘Excessive nitrate in groundwater has emerged as a serious environmental concern. The elevated nitrate concentration in drinking water causes a serious threat to public health on account of the possible transformation of nitrate to nitrite, which is one of the main predisposing factors of methemoglobinemia [1].
