贵州医科大学学报

1994, (03)

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底物浓度选择在酶Km值测定中的重要性
The Importance of Selecting a Right Substrate Concentration in the Km Assay of a Michaelis-Menten Enzyne

吴晓黎,陶建蜀
WuXiaoli;et al(Department of Clinical Biochemistry,Guiyang Medical College)

摘要(Abstract):

以胰蛋白酶对酪蛋白的Km值测定为例,讨论了底物浓度选择在酶Km值测定中的重要性。底物酪蛋白的浓度分为三组:高浓度(1~5mg/ml),低浓度(0.1~1mg/ml)和极低浓度(0.02~0.08mg/ml),高底物浓度(>8Km)的测定结果不能有效地区分多酶(作用于同一底物的多种酶)的存在,也不符合作图法要求的准确度,所得到的Km值可能是不准确的或错误的。因此,测定Km值时所用的底物浓度必须在0.2~5Km范围内,必要时应包括更广泛的浓度范围(包括高底物浓度)。
The data of Km assay of casein hydrolyzed by trypsin was taken was taken as a example for discussing the importance of selecting a right substrate concentrations in the Km assay of a Michaelis-Menten enzyme.Casein(as substrate of trypsin) was divided to three levels of high concentrations(1-5mg/ml),low concentrations (0.1-1mg/ml) and very low concentrations(0.02-0.08mg/ml).High substrate concentrations (>8Km) is not enough to prove the exist of a multi-enzymes system,(two or more than two enzymes which catalyze the same substrate) and to come to the precision require by a plotting.It may result in a mistake or incorrect Km.The substrate concentrations used to assay Km must be in 0.2-5Km which is in the levels of low and very low concentrations,if necessaray,a larger range of substrate concentrations should be selected that include high level.

关键词(KeyWords): 酪蛋白类,胰蛋白酶类,米氏常数
caseins;trypsin;Km;multi-enzymes

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 吴晓黎,陶建蜀
WuXiaoli;et al(Department of Clinical Biochemistry,Guiyang Medical College)

Email:

DOI: 10.19367/j.cnki.1000-2707.1994.03.003

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