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 language | English |
|---|---|
| Pages (from-to) | 749-752 |
| Number of pages | 4 |
| Journal | Journal of Antibiotics |
| Volume | 71 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2018 Aug 1 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Pharmacology
- Drug Discovery
Cite this
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 journal › Article
}
TY - JOUR
T1 - Identification and heterologous expression of the actinoallolide biosynthetic gene cluster
AU - Inahashi, Yuki
AU - Shiraishi, Taro
AU - Take, Akira
AU - Matsumoto, Atsuko
AU - Takahashi, Yōko
AU - Ōmura, Satoshi
AU - Kuzuyama, Tomohisa
AU - Nakashima, Takuji
PY - 2018/8/1
Y1 - 2018/8/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85045735867&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85045735867&partnerID=8YFLogxK
U2 - 10.1038/s41429-018-0057-8
DO - 10.1038/s41429-018-0057-8
M3 - Article
AN - SCOPUS:85045735867
VL - 71
SP - 749
EP - 752
JO - Journal of Antibiotics
JF - Journal of Antibiotics
SN - 0021-8820
IS - 8
ER -