Practice in NADPH cofactor application in on-line drug biotransformation by capillary electrophoresis
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| Year of publication | 2015 |
| Type | Conference abstract |
| MU Faculty or unit | |
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| Description | Drug biotransformation performed by cytochrome P450 enzymes requires certain conditions to achieve maximal catalytic activity. The essential factor is the presence of reduced nicotinamide cofactor, NADPH (ß-nicotinamide adenine dinucteotide 2´-phosphate) in sufficient amount. In general protocols NADPH regenerating system consisting of glucose 6-phosphate, glucose 6-phosphate dehydrogenase, NADP+ and MgCl2 is recommended but usage of NADPH itself simplifies the reaction mixture significantly. Fewer reagents in the mixture are more convenient for on-line enzymatic reaction performed by capillary electrophoresis (CE). However using NADPH solely evokes solving its stability issue since NADPH in aqueous solution must be available in active form for long time period in the on-line system: i) NADPH stability is lower than NADH and it is strongly dependent on pH, temperature, and ionic strength ; ii) reduced nicotinamide cofactors are particularly unstable in phosphate buffers and iii) such reagent solution must be handled with particular care. |
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