Characterization of the polyene macrolide P450 epoxidase from Streptomyces natalensis that converts de-epoxypimaricin into pimaricin

The Biochemical Journal
Marta V MendesJesús F Aparicio

Abstract

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 degrees 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.

References

Jun 1, 1996·Molecular Microbiology·A W Munro, J G Lindsay
Apr 17, 2003·Applied Microbiology and Biotechnology·J F AparicioS B Zotchev
Apr 20, 2004·Journal of Bacteriology·Nuria AntónJesús F Aparicio

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Citations

Mar 28, 2008·Applied Microbiology and Biotechnology·Bin ZhugeJian Zhuge
Aug 27, 2009·Applied Microbiology and Biotechnology·Domingo MiranzoFrancisco Malpartida
Aug 14, 2012·Applied Microbiology and Biotechnology·Hong ZongJian Zhuge
Apr 24, 2013·Enzyme and Microbial Technology·Sebastian PeterMartin Hofrichter
Oct 25, 2011·Chemical Reviews·Christopher J ThibodeauxHung-wen Liu
Mar 27, 2012·Journal of the American Chemical Society·Santhosh SivaramakrishnanPaul R Ortiz de Montellano
Mar 28, 2008·Journal of the American Chemical Society·Yousong DingDavid H Sherman
Dec 6, 2008·Journal of the American Chemical Society·Heather D Johnson, Jon S Thorson
Jul 30, 2008·Natural Product Reports·Elizabeth S SattelyChristopher T Walsh
Apr 9, 2013·Bioscience, Biotechnology, and Biochemistry·Niamh StephensPatrick Caffrey
Oct 30, 2015·Applied Microbiology and Biotechnology·Jesús F AparicioJavier Santos-Aberturas
Sep 19, 2006·Biochimica Et Biophysica Acta·Frank HannemannRita Bernhardt
Feb 27, 2015·Applied Microbiology and Biotechnology·Cláudia M VicenteJesús F Aparicio
Mar 24, 2006·FEMS Microbiology Letters·Lorena L EnríquezJesús F Aparicio
Nov 23, 2016·Molecular Microbiology·Tan-Jun WangYong-Quan Li
Jul 18, 2008·Molecular BioSystems·Jesús F Aparicio, Juan F Martín
Sep 18, 2008·Chembiochem : a European Journal of Chemical Biology·Changsheng ZhangJon S Thorson
Dec 24, 2019·Current Protein & Peptide Science·Xin XieHui Jiang
Jul 11, 2020·International Journal of Molecular Sciences·Fanele Cabangile MnguniKhajamohiddin Syed
May 24, 2021·Journal of Biological Engineering·Guang ChenHuanzhang Xia
May 11, 2017·Biochemistry·Drew R TietzThomas C Pochapsky

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