NEW METHOD FOR GUANASE ACTIVITY MEASUREMENT BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

被引：6

作者：

CANEPARI, S ^{[1
]}

CARUNCHIO, V ^{[1
]}

GIRELLI, AM ^{[1
]}

MESSINA, A ^{[1
]}

机构：

[1] UNIV ROMA LA SAPIENZA,DIPARTIMENTO CHIM,P ALDO MORO 5,I-00185 ROME,ITALY

来源：

JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS | 1993年 / 616卷 / 01期

关键词：

D O I：

10.1016/0378-4347(93)80467-I

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A rapid isocratic high-performance liquid chromatographic method for the determination of guanase (EC 3.5.4.3) activity is proposed. The method is highly reproducible, with a coefficient of variation of less than 1%, and requires only ca. 10 min for a complete chromatographic separation of the enzyme reaction mixture. The method allows the detection of nanomolar changes in the concentrations of both the substrate and the product, and does not require additional reactions or sample pretreatment. Kinetic studies with the proposed method showed the guanase activity to have an apparent Michaelis constant of 13.3 and 8.5 muM, and a maximum rate of 1.95 and 3.84 pmol/min per mg protein at 37-degrees-C, in Tris-HCl and phosphate buffer, respectively.

引用

页码：25 / 30

页数：6

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