Abstract: This chapter discusses the determination of formate dehydrogenase. The assay depends on measurement of the rate of increase of optical density at 340 mμ consequent on the reduction of diphosphopyridine nucleotide (DPN) in the presence of formate. The complete reaction system in a 1.5 ml silica cell (1 cm light path) consists of 0.25 ml of phosphate buffer, 0.25 ml of sodium formate, 0.1 ml of DPN, enzyme extract, and water to a total volume of 1 ml. The blank cell lacks formate and DPN. The reaction is started by addition of enzyme to both cells, and the rate of increase of extinction at 340 mμ is measured. Enzyme may be placed initially in both test and blank cell and the reaction be started by adding formate to the test cell. One unit of enzyme is defined as the amount of enzyme that catalyzes the reduction of 1 micromole of DPN per minute under the conditions of assay. Specific activity is expressed as units of enzyme per milligram of protein. The purified enzyme is specific for formate: no activity has been detected with methanol, ethanol, formaldehyde, oxalate, acetate, pyruvate, or malate.
Publication Year: 1966
Publication Date: 1966-01-01
Language: en
Type: book-chapter
Indexed In: ['crossref']
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Cited By Count: 48
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