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Synergistic Actions of Tailoring Enzymes in Pradimicin Biosynthesis
Author(s) -
Napan Kandy,
Zhang Shuwei,
Morgan Whitney,
Anderson Thomas,
Takemoto Jon Y.,
Zhan Jixun
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402306
Subject(s) - biosynthesis , enzyme , chemistry , streptomyces coelicolor , biochemistry , stereochemistry , cytochrome p450 , natural product , gene , mutant
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.