Abstract
Lankacidin is a unique 17-membered macrocyclic antibiotic different from usual even-membered macrolides. Based on the gene organization of the lankacidin biosynthetic cluster coded on the linear plasmid pSLA2-L in Streptomyces rochei, we previously proposed a hypothesis of modular-iterative mixed polyketide biosynthesis for lankacidin. Two experimental evidences in this paper further strengthened this hypothesis. Heterologous expression of the lankacidin cluster (lkcA-lkcO) in Streptomyces lividans resulted in lankacidinol A production, indicating that the gene cluster is sufficient for the synthesis of the lankacidin skeleton. In addition, a gene fusant of lkcF and lkcG produced lankacidin at a similar level to the parent strain, suggesting that an iterative function of the LkcF protein is unlikely. These results are consistent with the hypothesis that LkcC is used four times and LkcA, LkcF and LkcG are used modularly to accomplish eight condensation reactions leading to the lankacidin skeleton.
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References
Staunton J, Weissman KJ . Polyketide biosynthesis: a millennium review. Nat Prod Rep 18: 380–416 ( 2001)
Wilkinson B, Foster G, Rudd BAM, Taylor NL, Blackaby AP, Sidebottom PJ, Cooper DJ, Dawson MJ, Buss AD, Gaisser S, Böhm IU, Rowe CJ, Cortés J, Leadlay PF, Staunton J . Novel octaketide macrolides related to 6-deoxyerythronolide B provide evidence for iterative operation of the erythromycin polyketide synthase. Chem Biol 7: 111–117 ( 2000)
Hardt IH, Steinmetz H, Gerth K, Sasse F, Reichenbach H, Höfle G . New natural epothilones from Sorangium cellulosum, strains So ce90/B2 and So ce90/D13: Isolation, structure elucidation, and SAR studies. J Nat Prod 64: 847–856 ( 2001)
Gaitatzis N, Silakowski B, Kunze B, Nordsiek G, Blöker H, Höfle G, Müller R . The biosynthesis of the aromatic myxobacterial electron transport inhibitor stigmatellin is directed by a novel type of modular polyketide synthase. J Biol Chem 277: 13082–13090 ( 2002)
Olano C, Wilkinson B, Moss SJ, Sheridan R, Braña AF, Méndez C, Leadlay PF, Salas JA . Evidence from engineered gene fusions for the repeated use of a module in a modular polyketide synthase. Chem Commun 2003: 2780–2782 ( 2003)
He J, Hertweck C . Functional analysis of the aureothin iterative type I polyketide synthase. ChemBioChem 6: 908–912 ( 2005)
Kinashi H, Mori E, Hatani A, Nimi O . Isolation and characterization of large linear plasmids from lankacidin-producing Streptomyces species. J Antibiot 47: 1447–1455 ( 1994)
Kinashi H, Fujii S, Hatani A, Kurokawa T, Shinkawa H . Physical mapping of the linear plasmid pSLA2-L and localization of the eryAI and actI homologs. Biosci Biotechnol Biochem 62: 1892–1897 ( 1998)
Mochizuki S, Hiratsu K, Suwa M, Ishii T, Sugino F, Yamada K, Kinashi H . The large linear plasmid pSLA2-L of Streptomyces rochei has an unusually condensed gene organization for secondary metabolism. Mol Microbiol 48: 1501–1510 ( 2003)
Arakawa K, Sugino F, Kodama K, Ishii T, Kinashi H . Cyclization mechanism for the synthesis of macrocyclic antibiotic lankacidin in Streptomyces rochei. Chem Biol 12: 249–256 ( 2005)
Yu TW, Müller R, Müller M, Zhang X, Draeger G, Kim CG, Leistner E, Floss HG . Mutational analysis and reconstituted expression of the biosynthetic genes involved in the formation of 3-amino-5-hydroxybenzoic acid, the starter unit of rifamycin biosynthesis in Amycolatopsis mediterranei S699. J Biol Chem 276: 12546–12555 ( 2001)
Fernández-Moreno MA, Caballero J, Hopwood DA, Malpartida F . The act cluster contains regulatory and antibiotic export genes, direct targets for translational control by the bldA tRNA gene of Streptomyces. Cell 66: 769–780 ( 1991)
McDaniel R, Ebert-Khosla S, Hopwood DA, Khosla C . Engineered biosynthesis of novel polyketides. Science 262: 1546–1550 ( 1993)
Bierman M, Logan R, O'Brien K, Seno ET, Rao RN, Schoner BE . Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116: 43–49 ( 1992)
McDaniel R, Kao CM, Hwang SJ, Khosla C . Engineered intermodular and intramodular polyketide synthase fusions. Chem Biol 4: 667–674 ( 1997)
Ishikawa J, Niino Y, Hotta K . Construction of pRES18 and pRES19, Streptomyces-Escherichia coli shuttle vectors carrying multiple cloning sites. FEMS Microbiol Lett 145: 113–116 ( 1996)
Hopwood DA . Genetic contributions to understanding polyketide synthases. Chem Rev 97: 2465–2497 ( 1997)
Huang G, Zhang L, Birch RG . A multifunctional polyketide-peptide synthetase essential for albicidin biosynthesis in Xanthomonas albilineans. Microbiology 147: 631–642 ( 2001)
Mayorga ME, Timberlake WE . The developmentally regulated Aspergillus nidulans wA gene encodes a polypeptide homologous to polyketide and fatty acid synthases. Mol Gen Genet 235: 205–212 ( 1992)
Fujii I, Watanabe A, Sankawa U, Ebizuka Y . Identification of Claisen cyclase domain in fungal polyketide synthase WA, a naphthopyrone synthase of Aspergillus nidulans. Chem Biol 8: 189–197 ( 2001)
Yu JH, Leonard TJ . Sterigmatocystin biosynthesis in Aspergillus nidulans requires a novel type I polyketide synthase. J Bacteriol 177: 4792–4800 ( 1995)
Rahman AS, Hothersall J, Crosby J, Simpson TJ, Thomas CM . Tandem duplicated acyl carrier proteins, which increase polyketide antibiotic production, can apparently function either in parallel or in series. J Biol Chem 280: 6399–6408 ( 2005)
Sambrook J, Fritsch EF, Maniatis T . In Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York, ( 1989)
Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA . In Practical Streptomyces Genetics. The John Innes Foundation, Norwich, ( 2000)
Zhang H, Shinkawa H, Ishikawa J, Kinashi H, Nimi O . Improvement of transformation system in Streptomyces using a modified regeneration medium. J Ferment Bioeng 83: 217–221 ( 1997)
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Tatsuno, S., Arakawa, K. & Kinashi, H. Analysis of Modular-iterative Mixed Biosynthesis of Lankacidin by Heterologous Expression and Gene Fusion. J Antibiot 60, 700–708 (2007). https://doi.org/10.1038/ja.2007.90
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DOI: https://doi.org/10.1038/ja.2007.90
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