Microdetermination of glycerol using bacterial NADH-linked luciferase

A bioluminescent assay for determination of glycerol using bacterial NADH-linked luciferase was developed and successfully applied to measurement of glycerol release from human fat cells. The procedure is based on enzymic conversion of glycerol to 3-phosphoglycerate, which is irreversible in the presence of arsenate, and subsequent determination of NADH formed in the glycerol-phosphate- and glyceraldehyde-3-phosphate dehydrogenase reactions respectively. Bioluminescent determination of glycerol is about 100 times more sensitive than conventional spectrophotometry, thus permitting more than 100 determinations to be carried out on needle biopsy specimens.