A Rapid Enzymatic Assay for Glycerol
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TL;DR
An alternative and more rapid method has been developed which is based on the following reactions: glycerol + ATP + L-glycerol 1-phosphate + ADP (glycersol kinase) ADP + phosphoenolpyruvate + NADH2 (lactate dehydrogenase)
Abstract
ENZYME methods for the assay of glycerol using glycerol dehydrogenase1 or glycerol kinase with L-glycerol 1 -phosphate dehydrogenase2 and based on the reduction of NAD to NADH2 (measured by change in extinction at 340 mµ) have the disadvantage that the reaction time is comparatively long (about 30 min). An alternative and more rapid method has been developed which is based on the following reactions: glycerol + ATP ⇌ L-glycerol 1-phosphate + ADP (glycerol kinase) ADP + phosphoenolpyruvate ⇌ ATP + pyruvate (pyruvate kinase) pyruvate + NADH2 ⇌ lactate + NAD (lactate dehydrogenase)