Peroxide-driven catalysis of the heme domain of A. radioresistens cytochrome P450 116B5 for sustainable aromatic rings oxidation and drug metabolites production.

Peroxide-driven catalysis of the heme domain of A. radioresistens cytochrome P450 116B5 for sustainable aromatic rings oxidation and drug metabolites production. N Biotechnol. 2019 Aug 29;: Authors: Ciaramella A, Catucci G, Di Nardo G, Sadeghi SJ, Gilardi G Abstract The heme domain of cytochrome P450 116B5 from Acinetobacter radioresistens (P450 116B5hd), a self-sufficient class VII P450, was functionally expressed in Escherichia coli, purified and characterised in active form. Its unusually high reduction potential (-144 ± 42 mV) and stability in the presence of hydrogen peroxide make this enzyme a good candidate for driving catalysis with the so-called peroxide shunt, avoiding the need for a reductase and the expensive cofactor NAD(P)H. The enzyme is able to carry out the peroxide-driven hydroxylation of aromatic compounds such as p-nitrophenol (KM = 128.85 ± 29.51 μM and kcat = 2.65 ± 0.14 min-1), 10-acetyl-3,7-dihydroxyphenoxazine (KM = 6.01 ± 0.32 μM and kcat = 0.33 ± 0.03 min-1), and 3,5,3',5'tetramethylbenzidine (TMB). Moreover, it catalyses different reactions on well-known drugs such as hydroxylation of diclofenac (KM = 49.60 ± 6.30 μM and kcat = 0.06 ± 0.01 min-1) and N-desmethylation of tamoxifen (KM = 57.20 ± 7.90 μM and kcat = 0.79 ± 0.04 min-1). The data demonstrate that P450 116B5hd is an efficient biocatalyst for sustainable app...
Source: New Biotechnology - Category: Biotechnology Authors: Tags: N Biotechnol Source Type: research