Correcting a potential defect in an enzymatic cycle for NADP.
An enzymatic cycle for NADP which uses as one of its enzymes glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides has occasionally caused trouble due to failure to completely heat-kill this enzyme before the indicator step. It was found that a very small increase in pH was the cause of this. It was also found that the other two proteins present in the reagent greatly increase heat inactivation of the enzyme. The inactivation problem is completely overcome by keeping the pH below 7.2.
D005470:Fluorometry / Q000379:methods; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors; D005969:Glutamate Dehydrogenase; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007934:Leuconostoc / Q000201:enzymology*; D009249:NADP / Q000032:analysis*; D011092:Polyethylene Glycols; D012710:Serum Albumin, Bovine
D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; D009249:NADP; D005954:Glucosephosphate Dehydrogenase; D005969:Glutamate Dehydrogenase