title: Highly Efficient CYP167A1 (EpoK) dependent Epothilone B Formation and Production of 7-Ketone Epothilone D as a New Epothilone Derivative.
abstract: Since their discovery in the soil bacterium Sorangium cellulosum, epothilones have emerged as a valuable substance class with promising anti-tumor activity. Because of their benefits in the treatment of cancer and neurodegenerative diseases, epothilones are targets for drug design and pharmaceutical research. The final step of their biosynthesis - a cytochrome P450 mediated epoxidation of epothilone C/D to A/B by CYP167A1 (EpoK) - needs significant improvement, in particular regarding the efficiency of its redox partners. Therefore, we have investigated the ability of various hetero- and homologous redox partners to transfer electrons to EpoK. Hereby, a new hybrid system was established with conversion rates eleven times higher and Vmax of more than seven orders of magnitudes higher as compared with the previously described spinach redox chain. This hybrid system is the most efficient redox chain for EpoK described to date. Furthermore, P450s from So ce56 were identified which are able to convert epothilone D to 14-OH, 21-OH, 26-OH epothilone D and 7-ketone epothilone D. The latter one represents a novel epothilone derivative and is a suitable candidate for pharmacological tests. The results revealed myxobacterial P450s from S. cellulosum So ce56 as promising candidates for protein engineering for biotechnological production of epothilone derivatives.
authors.affiliation: Department of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Fredy
authors.initials: F
authors.lastname: Kern
authors.affiliation: Institute of Bioanalytical Chemistry, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Tobias K F
authors.initials: TK
authors.lastname: Dier
authors.affiliation: Department of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Yogan
authors.initials: Y
authors.lastname: Khatri
authors.affiliation: Department of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Kerstin M
authors.initials: KM
authors.lastname: Ewen
authors.affiliation: Unité Mixte de Recherches, 1136 Interaction arbres microorganismes INRA, Nancy University, 54506 Vandoeuvre-lès-Nancy cedex, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Jacquot
authors.affiliation: Institute of Bioanalytical Chemistry, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Dietrich A
authors.initials: DA
authors.lastname: Volmer
authors.affiliation: Department of Biochemistry, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Rita
authors.initials: R
authors.lastname: Bernhardt
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004579:Electron Transport; D004583:Electrons; D034261:Epothilones / Q000096:biosynthesis / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005288:Ferredoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007659:Ketones / Q000737:chemistry / Q000378:metabolism; D009231:Myxococcales / Q000737:chemistry / Q000201:enzymology*; D010084:Oxidation-Reduction; D010802:Phylogeny; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D034261:Epothilones; D005288:Ferredoxins; D007659:Ketones; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System