Synergistic actions of tailoring enzymes in pradimicin biosynthesis.

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. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Citation

Kandy Napan, Shuwei Zhang, Whitney Morgan, Thomas Anderson, Jon Y Takemoto, Jixun Zhan. Synergistic actions of tailoring enzymes in pradimicin biosynthesis. Chembiochem : a European journal of chemical biology. 2014 Oct 13;15(15):2289-96

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PMID: 25155298

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