Abstract
Actinoallolides are anti-trypanosomal macrolides isolated from the secondary metabolites of two endophytic actinomycete strains, Actinoallomurus fulvus MK10-036 and K09-0307. A putative actinoallolide biosynthetic gene cluster was predicted from the genome sequence of the strain K09-0307. The gene cluster spans a contiguous 53 kb DNA region that comprises seven genes encoding three PKSs (aalA1, aalA2, and aalA3), cytochrome P450 (aalB), acyl-CoA dehydrogenase (aalC), crotonyl-CoA reductase (aalD), and TetR family regulator (aalR). The entire gene cluster was cloned into a plasmid pYIK1 by assembling DNA fragments, which were obtained from two cosmids containing left and right parts of the gene cluster. Following the introduction of an ermE* promoter at 100bp upstream from the start codon of aalA1, the gene cluster was introduced into Streptomyces coelicolor M1152. Subsequent LC-MS analysis revealed production of actinoallolide A in the culture broth. Thus, the actinoallolide biosynthetic gene cluster was identified by heterologous expression in Streptomyces.
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Acknowledgements
We are grateful to Prof. Gregory L. Challis (University of Warwick, UK) for providing E. coli ET12567 and the John Innes Centre (UK) for providing Streptomyces coelicolor M1152 and plasmids pUB307 and pIJ10702. We also thank Dr. Kaia Palm (Protobios LLC, Estonia) for helpful experimental advice. This study was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers 15K18888 to Y.I. and 16H06453 to T.K., a Kitasato University Research Grant for Young Researchers (Y.I.), and funds from the Institute for Fermentation, Osaka (IFO) and from JSPS through the “Funding Program for Next Generation World-Leading Researchers (NEXT Program)” (LS028 to T.K.), which was initiated by the Council for Science and Technology Policy (CSTP), Japan.
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Inahashi, Y., Shiraishi, T., Také, A. et al. Identification and heterologous expression of the actinoallolide biosynthetic gene cluster. J Antibiot 71, 749–752 (2018). https://doi.org/10.1038/s41429-018-0057-8
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DOI: https://doi.org/10.1038/s41429-018-0057-8
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