RT info:eu-repo/semantics/article
T1 Characterization of the polyene macrolide P450 epoxidase from Streptomyces natalensis that converts de-epoxypimaricin into pimaricin
A1 Mendes, Marta V.
A1 Antón Fidalgo, Nuria
A1 Martín, Juan F.
A1 Aparicio, Jesús F.
K1 Cytochrome P450 mono-oxygenase
K1 Epoxidase
K1 Pimaricin
K1 Polyene
K1 Polyketide synthase
K1 Antibióticos
K1 Antibiotics
K1 Biosíntesis
K1 Biosynthesis
AB The biosynthesis of the antifungal agent pimaricin by Streptomyces natalensis has been proposed to involve a cytochrome P450 encoded by the gene pimD. Pimaricin is derived from its immediate precursor de-epoxypimaricin by epoxidation of the C-4–C-5 double bond on the macrolactone ring. We have overproduced PimD with a N-terminal His6 affinity tag in Escherichia coli and purified the enzyme for kinetic analysis. The protein showed a reduced CO-difference spectrum with a Soret maximum at 450 nm, indicating that it is a cytochrome P450. Purified PimD was shown to catalyse the in vitro C-4–C-5 epoxidation of 4,5-de-epoxypimaricin to pimaricin. The enzyme was dependent on NADPH for activity with optimal pH at 7.5, and the temperature optimum was 30 °C. The kcat value for the epoxidation of de-epoxypimaricin was similar to the values reported for other macrolide oxidases. Enzyme activity was inhibited at high substrate concentration. This is the first time that a polyene macrolide P450 mono-oxygenase has been expressed heterologously and studied. The unique specificity of this epoxidase should be useful for the oxidative modification of novel polyene macrolide antibiotics.
PB Biochemical society
SN 0264-6021
YR 2005
FD 2005
LK https://hdl.handle.net/10259/12148
UL https://hdl.handle.net/10259/12148
LA eng
DS Repositorio Institucional de la Universidad de Burgos
RD 03-oct-2026