* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
With 1H-imidazole; Mn(TDCPP)Cl; dihydrogen peroxide In dichloromethane; acetonitrile at 20℃; for 4h;
Multi-step reaction with 5 steps
1: 55 percent / NBS, benzoyl peroxide / CCl4 / 6 h / Heating
2: LiBr / dimethylformamide / 5 h / 80 - 100 °C
3: m-chloroperbenzoic acid / CH2Cl2 / 2 h / 35 °C
4: 1.) potassium tert-butoxide, 2.) HCl / 1.) THF, 15 deg C, 2 h; 2.) THF, 15 deg C, 20 min
5: 100 percent / H2 / Pd/C / tetrahydrofuran / 1.5 h / 2585.7 Torr / Ambient temperature
With β-nicotinamide adenine dinucleotide 2'-phosphate reduced tetra(cyclohexylammonium) salt; cytochrome P450 BM3 from Bacillus megaterium Asp251Gly/Gln307His mutant at 20℃; aq. phosphate buffer; Enzymatic reaction;
With Vitamin C; Coprinellus radians peroxygenase; dihydrogen peroxide at 20℃; for 0.05h; aq. phosphate buffer; Enzymatic reaction; regioselective reaction;
With glucose-6-phosphate dehydrogenase; glucose-6-phosphate; ferredoxin 8; ferredoxin-NADP<SUP>+</SUP> reductase B; recombinant CYP267B1 from Sorangium cellulosum So ce56; NADPH In ethanol at 30℃; for 3h; Enzymatic reaction;
In Vitro Conversions.
General procedure: A reconstituted in vitro system containing the corresponding P450 (0.5 mM), FdR_B (1.5 mM), and Fdx8 (10 mM), and acofactor regenerating system with glucose-6-phosphate (5 mM) and glucose-6-phosphate dehydrogenase (2 U/ml) in potassium phosphate buffer (20 mM, pH 7.4,1% glycerin) was used. The potential substrates, except olanzapine and omeprazole(both dissolved in dimethylsulfoxide),were dissolved in ethanol (10mM) and addedto an end concentration of 200 mM. The total volume of the reaction was 250 ml.The reaction was started by the addition of NADPH (800 mM) and carried out for3 hours at 30C. For substrates 1, 9, 11-15, and 18-21, 1M glycine buffer (pH 11)or acetate buffer (pH 4) was added after reaction to enable improved recovery of theanalytes. Therefore, the reaction was stopped by adding buffer (300 ml) ororganic solvent (500 ml). The extraction was performed twice with 500 ml of theappropriate solvent (see (Supplemental Table 0). A negative control withoutP450 was implemented for each substrate to verify the P450-dependent reaction.
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