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Redox Coenzymes
No.165(April 2015)
Enzymatic cycling methods have been designed for amplifying sensitivity to assay a small amount of substrate or enzyme activity.1) Earlier studies on enzymatic cycling methods were described on DAN+, NADP+, and CoA.2-4) Thio-NAD+ (1) contributed to improvement of the enzymatic cycling method.1,5)
Conversion of MVA to HMG-CoA by HMG-CoA reductase6)
An example of application of 1 for the enzymatic cycling method is shown above. As 1 is converted to thio-NADH (2) by such an enzymatic reaction, the reaction can be traced by the production of 2. Accumulation of 2 can be monitored by the increase in its absorbance (400 nm), which is at a longer wavelength than NADH (340nm).8) On the other hand, 2 was also applied for analysis of the active form of bleomycin.9)
References
- 1)Carnitine determination by an enzymatic cycling method with carnitine dehydrogenase
- 2)An enzymatic cycling method for nicotinamide-adenine dinucleotide with malic and alcohol dehydrogenases
- 3)The measurement of pyridine nucleotides by enzymatic cycling
- 4)Distribution of enzymes between nucleus and cytoplasm of single nerve cell bodies
- 5)ALT reagent with thionicotinamide adenine dinucleotide
- 6)An ultrasensitive enzymatic method for measuring mevalonic acid in serum
- 7)Spectrophotometric method for the assay of steroid 5-reductase activity of rat liver and prostate microsome
- 8)Ultrasensitive enzyme-linked immunosorbent assay (ELISA) of proteins by combination with the thio-NAD cycling method
- 9)The redox state of activated bleomycin
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