Determination of magnesium ion by use of the coupled-enzyme method with pyruvate kinase and lactate dehydrogenase.
We developed a new convenient method for determination of magnesium ion concentrations by use of magnesium ion-dependent enzymes; pyruvate kinase and lactate dehydrogenase. This method is based on the determination of the reaction rate of pyruvate kinase which uses MgADP- as substrate. The reaction rate of pyruvate kinase is dependent upon the formation of the complex with ADP and magnesium ion and the amount of the complex is dependent upon that of magnesium ion. The reaction rate of pyruvate kinase can be easily and spectrophotometrically determined by using lactate dehydrogenase and NADH as the decreased amount of absorbance at 340 nm.
Shimofuruya
H
H
Department of Materials Science and Engineering, Suzuka College of Technology, Mie, Japan.
Nakamura
M
M
Suzuki
J
J
D000244:Adenosine Diphosphate / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000737:chemistry*; D008274:Magnesium / Q000032:analysis*; D009243:NAD / Q000737:chemistry; D010084:Oxidation-Reduction; D010728:Phosphoenolpyruvate / Q000737:chemistry; D011770:Pyruvate Kinase / Q000737:chemistry*; D013053:Spectrophotometry; D013552:Swine
D009243:NAD; D000244:Adenosine Diphosphate; D010728:Phosphoenolpyruvate; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D011770:Pyruvate Kinase; D008274:Magnesium