Identification of the biosynthetic gene cluster for the antibiotic polyketide L-155,175 in Streptomyces hygroscopicus
- Authors
- Kim, Eun Young; Han, Jae Woo; Lee, Jee Yeon; Kim, Beom Seok
- Issue Date
- Nov-2012
- Publisher
- SPRINGER
- Citation
- FOLIA MICROBIOLOGICA, v.57, no.6, pp.543 - 550
- Indexed
- SCIE
SCOPUS
- Journal Title
- FOLIA MICROBIOLOGICA
- Volume
- 57
- Number
- 6
- Start Page
- 543
- End Page
- 550
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/107165
- DOI
- 10.1007/s12223-012-0173-y
- ISSN
- 0015-5632
- Abstract
- The antibiotic L-155,175, a potent antiparasitic and antifungal compound, has an unusual structure involving 16-membered macrolides that contain a tetrahydropyran ring connected through a three-carbon linker chain. To identify the biosynthetic gene cluster for L-155,175, a genomic DNA library of Streptomyces hygroscopicus ATCC31955 was constructed and screened with a degenerate primer set designed from a conserved region of the ketosynthase (KS) domain. Sequence analysis of a fosmid clone, pEY1D8 (34 kb), revealed multiple open reading frames (ORFs) encoding type I polyketide synthase (PKS). To determine whether the cloned genes are involved in L-155,175 biosynthesis, a deletion mutant (1D8m) was generated by homologous recombination, in which the gene encoding the KS domain was substituted with an apramycin-resistance gene by PCR-targeted Streptomyces gene replacement. LC-MS analysis showed that L-155,175 production was completely abolished in the 1D8m strain, thereby proving that the cloned gene is responsible for L-155,175 biosynthesis. The sequencing of two other fosmid clones (pEY8B10 and pEY1C9) harboring overlapping sequences from pEY1D8 revealed a 60-kb DNA segment encoding six ORFs for type I PKS harboring 12 modules. The domain organization of the PKS modules encoded by PKS exactly matched the structure of L-155,175. This is the first report on the gene cluster involved in the biosynthesis of L-155,175.
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Collections - Graduate School > Department of Plant Biotechnology > 1. Journal Articles
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