title: Synergistic actions of tailoring enzymes in pradimicin biosynthesis.
abstract: Three key tailoring enzymes in pradimicin biosynthesis: PdmJ, PdmW, and PdmN, were investigated. PdmW was characterized as the C-6 hydroxylase by structural characterization of the corresponding product, 6-hydroxy-G-2A. The efficiencies of the C-5 and C-6 hydroxylations, catalyzed respectively by PdmJ and PdmW, were low when they were expressed individually with the early biosynthetic enzymes that form G-2A. When these two cytochrome P450 enzymes were co-expressed, a dihydroxylated product, 5,6-dihydroxy-G-2A, was efficiently produced, indicating that these two enzymes work synergistically in pradimicin biosynthesis. Heterologously expressed PdmN in Streptomyces coelicolor CH999 converted G-2A to JX137a by ligating a unit of D-alanine to the carboxyl group. PdmN has relaxed substrate specificity toward both amino acid donors and acceptors. Through combinatorial biosynthesis, a series of new pradimicin analogues were produced.
authors.affiliation: Department of Biological Engineering, Utah State University, 4105 Old Main Hill, Logan, UT 84322 (USA).
authors.forename: Kandy
authors.initials: K
authors.lastname: Napan
authors.forename: Shuwei
authors.initials: S
authors.lastname: Zhang
authors.forename: Whitney
authors.initials: W
authors.lastname: Morgan
authors.forename: Thomas
authors.initials: T
authors.lastname: Anderson
authors.forename: Jon Y
authors.initials: JY
authors.lastname: Takemoto
authors.forename: Jixun
authors.initials: J
authors.lastname: Zhan
mesh_terms: D018943:Anthracyclines / Q000737:chemistry / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D008968:Molecular Conformation; D000095643:Pradimicins and Benanomicins
chemical_list: D018943:Anthracyclines; D003577:Cytochrome P-450 Enzyme System; C066995:pradimicin N; D000095643:Pradimicins and Benanomicins
keywords: combinatorial biosynthesis; cytochrome P450 hydroxylase; polyketides; pradimicin; synergistic actions