The CYP450 enzyme isolated from bacteria in presence of 20% glycerol, 1mM DTT, 1mM PMSF in 0.1M Potassium phosphate buffer pH 7.4, during assay with carbon monooxide i got the peak at 418nm and not at P450 why this happened ??? kindly suggest.
Hello, 50 per cent of the cytochrome P450 is found in its inactive form, P420. Glycerol is known to prevent inactivation of cytochrome P450 to P420 in microsomal membranes treated with SH-agents, detergents etc. and causes reconversion of P420 into P450 in the membranes. So, perhaps you could play around with glycerol concentrations to see what concentration could increase P450 concentrations for you.
Hello, The cytochrome P450 content in cells, which corresponded to the BPA degradation activity, gradually increased until the stationary phase and reached a maximum level at about 20 h.
The cytochrome P450 monooxygenase system is very unstable, and sodium dithionite prevented inactivation of the system, although sodium dithionite had an inhibitory effect on its activity. Also as mentioned in previous answer glycerol is effective stabilizer for their activities, but 10% solution.
Hope this info will help you good luck!
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