Abstract
Streptolydigin is a tetramic acid antibiotic produced by Streptomyces lydicus NRRL 2433 and involving a hybrid polyketide synthase (PKS)–nonribosomal peptide synthetase (NRPS) system in its biosynthesis. The streptolydigin amino-acid precursor, 3-methylaspartate, has been proposed to be condensed to the polyketide portion of the molecule by a NRPS composed by three enzymes (SlgN1, SlgN2 and SlgL). On the other hand, biosynthesis of the polyketide moiety involves the participation of cytochrome P450 SlgO2 for the correct cyclization of the characteristic bicyclic ketal. Independent disruption of slgN1, slgN2, slgL or slgO2 resulted in S. lydicus mutants unable to produce the antibiotic thus confirming the involvement of these genes in the biosynthesis of the antibiotic. These mutants did not accumulate any streptolydigin biosynthesis intermediate or shunt product derived from early polyketides released from the PKS. However, they produced three novel compounds identified as 4-(2-carboxy-propylamino)-3-chloro-benzoic acid, 4-(2-carboxy-propylamino)-3-hydroxy-benzoic acid and 4-(2-carboxy-propylamino)-benzoic acid, which were designated as christolane A, christolane B and christolane C, respectively. These compounds have been shown to exert some antibiotic activity.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Accession codes
References
Newman, D. J. & Cragg, G. M. Natural products as sources of new drugs over the last 25 years. J. Nat. Prod. 70, 461–477 (2007).
Butler, M. S. Natural products to drugs: natural product-derived compounds in clinical trials. Nat. Prod. Rep. 25, 475–516 (2008).
Cragg, G. M., Grothaus, P. G. & Newman, D. J. Impact of natural products on developing new anti-cancer agents. Chem. Rev. 109, 3012–3043 (2009).
Olano, C., Méndez, C. & Salas, J. A. Antitumor compounds from actinomycetes: from gene clusters to new derivatives by combinatorial biosynthesis. Nat. Prod. Rep. 26, 628–660 (2009).
Olano, C., Méndez, C. & Salas, J. A. Antitumor compounds from marine actinomycetes. Mar. Drugs 7, 210–248 (2009).
Staunton, J. & Weissman, K. J. Polyketide biosynthesis: a millennium review. Nat. Prod. Rep. 18, 380–416 (2001).
Walsh, C. T. Polyketide and nonribosomal peptide antibiotics: modularity and versatility. Science 303, 1805–1810 (2004).
Fischbach, M. A. & Walsh, C. T. Assembly-line enzymology for polyketide and nonribosomal Peptide antibiotics: logic, machinery, and mechanisms. Chem. Rev. 106, 3468–3496 (2006).
Sieber, S. A. & Marahiel, M. A. Molecular mechanisms underlying nonribosomal peptide synthesis: approaches to new antibiotics. Chem. Rev. 105, 715–738 (2005).
Du, L., Sánchez, C. & Shen, B. Hybrid peptide-polyketide natural products: biosynthesis and prospects toward engineering novel molecules. Metab. Eng. 3, 78–95 (2001).
Walsh, C. T. et al. Tailoring enzymes that modify nonribosomal peptides during and after chain elongation on NRPS assembly lines. Curr. Opin. Chem. Biol 5, 525–534 (2001).
Rix, U., Fischer, C., Remsing, L. L. & Rohr, J. Modification of post-PKS tailoring steps through combinatorial biosynthesis. Nat. Prod. Rep. 19, 542–580 (2002).
Yin, X., O'Hare, T., Gould, S. J. & Zabriskie, T. M. Identification and cloning of genes encoding viomycin biosynthesis from Streptomyces vinaceus and evidence for involvement of a rare oxygenase. Gene 312, 215–224 (2003).
Walsh, C. T. The chemical versatility of natural-product assembly lines. Acc. Chem. Res. 41, 4–10 (2008).
Olano, C., Méndez, C. & Salas, J. A. Post-PKS tailoring steps in natural product-producing actinomycetes from the perspective of combinatorial biosynthesis. Nat. Prod. Rep. 27, 571–616 (2010).
Schobert, R. & Schlenk, A. Tetramic and tetronic acids: an update on new derivatives and biological aspects. Bioorg. Med. Chem. 16, 4203–4221 (2008).
Sánchez-Hidalgo, M., Núñez, L. E., Méndez, C. & Salas, J. A. Involvement of the beta subunit of RNA polymerase in resistance to streptolydigin and streptovaricin in the producer organisms Streptomyces lydicus and Streptomyces spectabilis. Antimicrob. Agents Chemother 54, 1684–1692 (2010).
Olano, C. et al. Deciphering biosynthesis of the RNA polymerase inhibitor streptolydigin and generation of glycosylated derivatives. Chem. Biol 16, 1031–1044 (2009).
Horna, D. H. et al. Biosynthesis of the RNA polymerase inhibitor streptolydigin in Streptomyces lydicus: tailoring modification of 3-methyl-aspartate. J. Bacteriol 193, 2647–2651 (2011).
Gómez, C. et al. Amino acid precursor supply in the biosynthesis of the RNA polymerase inhibitor streptolydigin by Streptomyces lydicus. J. Bacteriol 193, 4214–4223 (2011).
Kieser, T., Bibb, M. J., Buttner, M. J., Chater, K. F. & Hopwood, D. A. Practical Streptomyces Genetics, The John Innes Foundation: Norwich, (2000).
Fernández, E. et al. Identification of two genes from Streptomyces argillaceus encoding glycosyltransferases involved in transfer of a disaccharide during biosynthesis of the antitumor drug mithramycin. J. Bacteriol 180, 4929–4937 (1998).
Sambrook, J., Fritsch, E. F. & Maniatis, T. Molecular cloning: a laboratory manual, Cold Spring Harbour Laboratory press New York, Cold Spring Harbour, (1989).
Bierman, M. et al. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116, 43–49 (1992).
Olano, C. et al. Biosynthesis of the angiogenesis inhibitor borrelidin by Streptomyces parvulus Tü4055: cluster analysis and assignment of functions. Chem. Biol 11, 87–97 (2004).
Menéndez, N. et al. Deoxysugar transfer during chromomycin A3 biosynthesis in Streptomyces griseus subsp. griseus: new derivatives with antitumor activity. Appl. Environ. Microbiol 72, 167–177 (2006).
Tang, G. L., Cheng, Y. Q. & Shen, B. Leinamycin biosynthesis revealing unprecedented architectural complexity for a hybrid polyketide synthase and nonribosomal peptide synthetase. Chem. Biol. 11, 33–45 (2004).
Garneau, S., Dorrestein, P. C., Kelleher, N. L. & Walsh, C. T. Characterization of the formation of the pyrrole moiety during clorobiocin and coumermycin A1 biosynthesis. Biochemistry 44, 2770–2780 (2005).
Bihlmaier, C. et al. Biosynthetic gene cluster for the polyenoyltetramic acid α-lipomycin. Antimicrob. Agents Chemother 50, 2113–2121 (2006).
Carlson, J. C. et al. Identification of the tirandamycin biosynthetic gene cluster from Streptomyces sp. 307-9. ChemBioChem 11, 564–572 (2010).
Mo, X. et al. Cloning and characterization of the biosynthetic gene cluster of the bacterial RNA polymerase inhibitor tirandamycin from marine-derived Streptomyces sp. SCSIO1666. Biochem. Biophys. Res. Commun 406, 341–347 (2011).
Du, L., Sánchez, C., Chen, M., Edwards, D. J. & Shen, B. The biosynthetic gene cluster for the antitumor drug bleomycin from Streptomyces verticillus ATCC15003 supporting functional interactions between nonribosomal peptide synthetases and a polyketide synthase. Chem. Biol 7, 623–642 (2000).
Weber, T. et al. Molecular analysis of the kirromycin biosynthetic gene cluster revealed β-alanine as precursor of the pyridone moiety. Chem. Biol 15, 175–188 (2008).
Mendes, M. V., Antón, N., Martín, J. F. & Aparicio, J. F. Characterization of the polyene macrolide P450 epoxidase from Streptomyces natalensis that converts de-epoxypimaricin into pimaricin. Biochem. J. 386, 57–62 (2005).
Shah, S. et al. Cloning, characterization and heterologous expression of a polyketide synthase and P-450 oxidase involved in the biosynthesis of the antibiotic oleandomycin. J. Antibiot 53, 502–508 (2000).
Schultz, A. W. et al. Biosynthesis and structures of cyclomarins and cyclomarazines, prenylated cyclic peptides of marine actinobacterial origin. J. Am. Chem. Soc 130, 4507–4516 (2008).
Yu, T. W. et al. Direct evidence that the rifamycin polyketide synthase assembles polyketide chains processively. Proc. Natl Acad. Sci. USA 96, 9051–9056 (1999).
Doi-Katayama, Y. et al. Thioesterases and the premature termination of polyketide chain elongation in rifamycin B biosynthesis by Amycolatopsis mediterranei S699. J. Antibiot 53, 484–495 (2000).
Xu, J., Wan, E., Kim, C. J., Floss, H. G. & Mahmud, T. Identification of tailoring genes involved in the modification of the polyketide backbone of rifamycin B by Amycolatopsis mediterranei S699. Microbiology 151, 2515–2528 (2005).
Sun, Y. et al. ‘Streptomyces nanchangensis’, a producer of the insecticidal polyether antibiotic nanchangmycin and the antiparasitic macrolide meilingmycin, contains multiple polyketide gene clusters. Microbiology 148, 361–371 (2002).
Ou, X., Zhang, B., Zhang, L., Zhao, G. & Ding, X. Characterization of rrdA, a TetR family protein gene involved in the regulation of secondary metabolism in Streptomyces coelicolor. Appl. Environ. Microbiol. 75, 2158–2165 (2009).
Blankenfeldt, W. et al. Structure and function of the phenazine biosynthetic protein PhzF from Pseudomonas fluorescens. Proc. Natl Acad. Sci. USA 101, 16431–16436 (2004).
Van Lanen, S. G., Lin, S. & Shen, B. Biosynthesis of the enediyne antitumor antibiotic C-1027 involves a new branching point in chorismate metabolism. Proc. Natl Acad. Sci. USA 105, 494–499 (2008).
Barker, J. L. & Frost, J. W. Microbial synthesis of p-hydroxybenzoic acid from glucose. Biotechnol. Bioeng. 76, 376–390 (2001).
Stadthagen, G. et al. p-Hydroxybenzoic acid synthesis in Mycobacterium tuberculosis. J. Biol. Chem. 280, 40699–40706 (2005).
He, J. & Hertweck, C. Biosynthetic origin of the rare nitroaryl moiety of the polyketide antibiotic aureothin: involvement of an unprecedented N-oxygenase. J. Am. Chem. Soc 126, 3694–3695 (2004).
Van Kuilenburg, A. B., Stroomer, A. E., Van Lenthe, H., Abeling, N. G. & Van Gennip, A. H. New insights in dihydropyrimidine dehydrogenase deficiency: a pivotal role for beta-aminoisobutyric acid? Biochem. J. 379, 119–124 (2004).
Beck, Z. Q., Burr, D.A. & Sherman, D. H. Characterization of the β-methylaspartate-α-decarboxylase (CrpG) from the cryptophycin biosynthetic pathway. ChemBioChem 8, 1373–1375 (2007).
Acknowledgements
We thank Luz Valero Rustarazo from the Laboratory of Proteomics CIPF (network ProteoRed) for the MS analysis. This research was supported by a grant of the Spanish Ministry of Science and Innovation (BFU2006–00404) and Red Temática de Investigación Cooperativa de Centros de Cáncer (Ministry of Health, ISCIII-RETIC RD06/0020/0026) to JAS. We thank Obra Social Cajastur for financial support to Carlos Olano and the Spanish Ministry of Science and Innovation for PhD student fellowship (FPI) to Cristina Gómez. We also thank the Spanish Ministry of Science and Innovation (SAF2008–01845) and the Centro de Investigación Príncipe Felipe for their economical support.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on The Journal of Antibiotics website
Supplementary information
Rights and permissions
About this article
Cite this article
Gómez, C., Olano, C., Palomino-Schätzlein, M. et al. Novel compounds produced by Streptomyces lydicus NRRL 2433 engineered mutants altered in the biosynthesis of streptolydigin. J Antibiot 65, 341–348 (2012). https://doi.org/10.1038/ja.2012.37
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/ja.2012.37
Keywords
This article is cited by
-
The Combinatorial Biosynthesis of “Unnatural” Products with Polyketides
Transactions of Tianjin University (2018)
-
Roles of type II thioesterases and their application for secondary metabolite yield improvement
Applied Microbiology and Biotechnology (2014)
