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Functional characterization of O-methyltransferases used to catalyse site-specific methylation in the post-tailoring steps of pradimicin biosynthesis
- Title
- Functional characterization of O-methyltransferases used to catalyse site-specific methylation in the post-tailoring steps of pradimicin biosynthesis
- Authors
- Han, J. W.; Ng, B. G.; Sohng, J. K.; Yoon, Y. J.; Choi, G. J.; Kim, B. S.
- Ewha Authors
- 윤여준
- SCOPUS Author ID
- 윤여준
- Issue Date
- 2018
- Journal Title
- JOURNAL OF APPLIED MICROBIOLOGY
- ISSN
- 1364-5072
1365-2672
- Citation
- JOURNAL OF APPLIED MICROBIOLOGY vol. 124, no. 1, pp. 144 - 154
- Keywords
- aromatic polyketide; methyltransferase; O-methyltransferase; polyketide; pradimicin; pradimicin biosynthesis; pradimicinone
- Publisher
- WILEY
- Indexed
- SCIE; SCOPUS
- Document Type
- Article
- Abstract
- Aims: To identify the roles of the two O-methyltransferase homologous genes pdmF and pdmT in the pradimicin biosynthetic gene cluster of Actinomadura hibisca P157-2. Methods and Results: Pradimicins are pentangular polyphenol antibiotics synthesized by bacterial type II polyketide synthases (PKSs) and tailoring enzymes. Pradimicins are naturally derivatized by combinatorial O-methylation at two positions (i.e., 7-OH and 11-OH) of the benzo[alpha]naphthacenequinone structure. PdmF and PdmT null mutants (PFKO and PTKO) were generated. PFKO produced the 11-O-demethyl shunt metabolites 11-O-demethylpradimicinone II (1), 11-O-demethyl-7-methoxypradimicinone II (2), 11-O-demethylpradimicinone I (3) and 11-O-demethylpradimicin A (4), while PTKO generated the 7-O-demethyl derivatives pradimicinone II (5) and 7-hydroxypradimicin A (6). Pradimicinones 1, 2, 3, and 5 were fed to a heterologous host Escherichia coli harbouring expression plasmid pET-22b::pdmF or pET-28a::pdmT. PdmF catalysed 11-O-methylation of pradimicinones 1, 2, and 3 regardless of O-methylation at the C-7 position, while PdmT was unable to catalyse 7-O-methylation when the C-11 hydroxyl group was methylated (5). Conclusions: PdmF and PdmT were involved in 11-O- and 7-O-methylations of the benzo[alpha]naphthacenequinone moiety of pradimicin, respectively. Methylation of the C-7 hydroxyl group precedes methylation of the C-11 hydroxyl group in pradimicin biosynthesis. Significance and Impact of the Study: This is the first reported demonstration of the functions of PdmF and PdmT for regiospecific O-methylation, which contributes to better understanding of the post-PKS modifications in pradimicin biosynthesis as well as to rational engineering of the pradimicin biosynthetic machinery.
- DOI
- 10.1111/jam.13619
- Appears in Collections:
- 자연과학대학 > 화학·나노과학전공 > Journal papers
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