Replacing Commercial 6-Phosphofructokinase in an Online Pyrophosphate Detection Assay

Detection of pyrophosphate is important in quantifying enzyme activity, particularly adenylation

domain activity during non-ribosomal peptide synthesis. The previous development of an enzyme

coupled PPi/NADH assay allowed the measurement of such activity in an online fashion using

commercially available components. Now, with a key enzyme – 6-phosphofructokinase – no longer

available, we have screened and identified viable replacement enzymes that can be expressed in high

yield and that are far superior in activity to the now discontinued commercial product. This will

support the ability of groups to continue to use this established online assay for pyrophosphate

detection.