Identification and heterologous expression of the actinoallolide biosynthetic gene cluster

Yuki Inahashi, Taro Shiraishi, Akira Take, Atsuko Matsumoto, Yōko Takahashi, Satoshi Ōmura, Tomohisa Kuzuyama, Takuji Nakashima

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)749-752
Number of pages4
JournalJournal of Antibiotics
Volume71
Issue number8
DOIs
Publication statusPublished - 2018 Aug 1
Externally publishedYes

Fingerprint

Multigene Family
acyl-CoA dehydrogenase (NADP+)
Acyl-CoA Dehydrogenase
Streptomyces coelicolor
Cosmids
Initiator Codon
Actinobacteria
DNA
Macrolides
Streptomyces
Cytochrome P-450 Enzyme System
Plasmids
Genome
Genes

ASJC Scopus subject areas

  • Pharmacology
  • Drug Discovery

Cite this

Inahashi, Y., Shiraishi, T., Take, A., Matsumoto, A., Takahashi, Y., Ōmura, S., ... Nakashima, T. (2018). Identification and heterologous expression of the actinoallolide biosynthetic gene cluster. Journal of Antibiotics, 71(8), 749-752. https://doi.org/10.1038/s41429-018-0057-8

Identification and heterologous expression of the actinoallolide biosynthetic gene cluster. / Inahashi, Yuki; Shiraishi, Taro; Take, Akira; Matsumoto, Atsuko; Takahashi, Yōko; Ōmura, Satoshi; Kuzuyama, Tomohisa; Nakashima, Takuji.

In: Journal of Antibiotics, Vol. 71, No. 8, 01.08.2018, p. 749-752.

Research output: Contribution to journalArticle

Inahashi, Y, Shiraishi, T, Take, A, Matsumoto, A, Takahashi, Y, Ōmura, S, Kuzuyama, T & Nakashima, T 2018, 'Identification and heterologous expression of the actinoallolide biosynthetic gene cluster', Journal of Antibiotics, vol. 71, no. 8, pp. 749-752. https://doi.org/10.1038/s41429-018-0057-8
Inahashi Y, Shiraishi T, Take A, Matsumoto A, Takahashi Y, Ōmura S et al. Identification and heterologous expression of the actinoallolide biosynthetic gene cluster. Journal of Antibiotics. 2018 Aug 1;71(8):749-752. https://doi.org/10.1038/s41429-018-0057-8
Inahashi, Yuki ; Shiraishi, Taro ; Take, Akira ; Matsumoto, Atsuko ; Takahashi, Yōko ; Ōmura, Satoshi ; Kuzuyama, Tomohisa ; Nakashima, Takuji. / Identification and heterologous expression of the actinoallolide biosynthetic gene cluster. In: Journal of Antibiotics. 2018 ; Vol. 71, No. 8. pp. 749-752.
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