Reaktion #657269
ord-24d260030d7d47418fa126c453d3229e
Reaktionsgleichung
Reagenzien
Reaktionsbedingungen
Vorschrift
Measurements of conversion rate of formaldehyde to dihydroxyacetone were performed through the enzyme-coupled spectrophotometric assay described in Example 1, wherein the NADH consumption resulting from the reduction of dihydroxyacetone to glycerol is measured. Individual reactions contained 5 μM of the enzyme (BAL, FLS r3a, FLS r3b, FLS r3c, FLS r3d, FLS r4a, and FLS r4b), 0.1 mg/mL glycerol dehydrogenase, 1 mM NADH, 50 mM potassium phosphate buffer at pH 8.0, 0.2 mM thiamine diphosphate 2 mM MgSO4, and formaldehyde ranging from 0 to 50 mM. The change in absorbance at 340 nm was monitored, which resulted from the oxidation-reduction reaction of NADH to NAD and dihydroxyacetone to glycerol catalyzed by the coupling enzyme glycerol dehydrogenase. An increased rate of dihydroxyacetone production from formaldehyde by formolase enzymes resulted in an increased rate of NADH oxidation. The change in absorbance at 340 nm was converted into concentrations using an extinction coefficient of 6220 M−1 cm−1.