Peroxide-driven catalysis of the heme domain of A. radioresistens cytochrome P450 116B5 for sustainable aromatic rings oxidation and drug metabolites production.
N Biotechnol
; 54: 71-79, 2020 Jan 25.
Article
em En
| MEDLINE
| ID: mdl-31473254
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⯱â¯42â¯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 (KMâ¯=â¯128.85⯱â¯29.51 µM and kcatâ¯=â¯2.65⯱â¯0.14â¯min-1), 10-acetyl-3,7-dihydroxyphenoxazine (KMâ¯=â¯6.01⯱â¯0.32 µM and kcatâ¯=â¯0.33⯱â¯0.03â¯min-1), and 3,5,3',5'tetramethylbenzidine (TMB). Moreover, it catalyses different reactions on well-known drugs such as hydroxylation of diclofenac (KMâ¯=â¯49.60⯱â¯6.30 µM and kcatâ¯=â¯0.06⯱â¯0.01â¯min-1) and N-desmethylation of tamoxifen (KMâ¯=â¯57.20⯱â¯7.90 µM and kcatâ¯=â¯0.79⯱â¯0.04â¯min-1). The data demonstrate that P450 116B5hd is an efficient biocatalyst for sustainable applications in bioremediation and human drug metabolite production.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Oxazinas
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Peróxidos
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Acinetobacter
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Benzidinas
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Sistema Enzimático do Citocromo P-450
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Biocatálise
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Nitrofenóis
Idioma:
En
Revista:
N Biotechnol
Assunto da revista:
BIOLOGIA MOLECULAR
/
ENGENHARIA BIOMEDICA
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Itália