A sensitive and rapid method for determination of pyrophosphatase activity.
A new, rapid and sensitive method of measuring the rate and amount of PPi hydrolysis is described. The method is based on registration of small pH changes, caused by decrease of hydrogen-ion concentration during the pyrophosphatase reaction. With this detected value of hydrogen-ion concentration change and the buffering capacity of each reaction mixture determined by titration, the rate and amount of PPi hydrolysis can be calculated. The results obtained by pH method are in good agreement with values determined by direct phosphate analysis. The described method can be applied to the continuous determination of pyrophosphatase activity in chromatophores, mitochondria, crude extracts or purified enzymes.
D020130:Bacterial Chromatophores / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D011755:Pyrophosphatases / Q000032:analysis*; D012247:Rhodospirillum rubrum / Q000201:enzymology
D011755:Pyrophosphatases