A continuous spectrophotometric assay for the enzymatic marker glucose 6-phosphatase

A continuous spectrophotometric assay for glucose 6-phosphatase is described. The method uses glucose dehydrogenase and mutarotase as ancillary enzymes. Glucose 6-phosphatase activity is measured by following NADH formation at 340 nm. The method is linear, at least up to 38 mU in the test which corresponds to a delta E of 0.24 min-1, when the enzyme is assayed in a microsomal fraction. We also discuss the method's suitability for subcellular fractionation. No other continuous assay for this important enzymatic marker of the endoplasmic reticulum is currently available.