Allantoin catabolism influences the production of antibiotics in Streptomyces coelicolor
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Purines are a primary source of carbon and nitrogen in soil; however, their metabolism is poorly understood in Streptomyces. Using a combination of proteomics, metabolomics, and metabolic engineering, we characterized the allantoin pathway in Streptomyces coelicolor. When cells grew in glucose minimal medium with allantoin as the sole nitrogen source, quantitative proteomics identified 38 enzymes upregulated and 28 downregulated. This allowed identifying six new functional enzymes involved in allantoin metabolism in S. coelicolor. From those, using a combination of biochemical and genetic engineering tools, it was found that allantoinase (EC 126.96.36.199) and allantoicase (EC 188.8.131.52) are essential for allantoin metabolism in S. coelicolor. Metabolomics showed that under these growth conditions, there is a significant intracellular accumulation of urea and amino acids, which eventually results in urea and ammonium release into the culture medium. Antibiotic production of a urease mutant strain showed that the catabolism of allantoin, and the subsequent release of ammonium, inhibits antibiotic production. These observations link the antibiotic production impairment with an imbalance in nitrogen metabolism and provide the first evidence of an interaction between purine metabolism and antibiotic biosynthesis.
KeywordsStreptomyces Allantoin Ammonium Antibiotic regulation
This work was supported by ANPCyT grants PICT 2007–00711 to PC and PICT2008-644 to HG, Fundación Perez-Guerrero grant to ER and PIP 100764 from CONICET to ER. ER, PC, and HG are members of the Research Career and LN is a doctoral fellow of CONICET. CLC is a doctoral fellow of CONACYT. We kindly thank Dr. Alun Jones and Dr. Amanda Nouwens for LC-MS assistance and David Hopwood for helpful comments. All proteomics work was performed at the proteomics facility at IMB and/or SCMB. We thank Paul Dyson (Swansea University) for kindly providing the derivative cosmids carrying transposon insertions and Monica Hourcade (Universidad Nacional de Rosario) for technical assistance in the metabolomic analysis.
Conflict of interest
The authors declare no competing financial interests.
- Ashiuchi M, Misono H (1999) Biochemical evidence that Escherichia coli hyi (orf b0508, gip) gene encodes hydroxypyruvate isomerase. Biochim Biophys Acta 1435(1–2):153–159Google Scholar
- Bentley SD, Chater KF, Cerdeno-Tarraga AM, Challis GL, Thomson NR, James KD, Harris DE, Quail MA, Kieser H, Harper D, Bateman A, Brown S, Chandra G, Chen CW, Collins M, Cronin A, Fraser A, Goble A, Hidalgo J, Hornsby T, Howarth S, Huang CH, Kieser T, Larke L, Murphy L, Oliver K, O’Neil S, Rabbinowitsch E, Rajandream MA, Rutherford K, Rutter S, Seeger K, Saunders D, Sharp S, Squares R, Squares S, Taylor K, Warren T, Wietzorrek A, Woodward J, Barrell BG, Parkhill J, Hopwood DA (2002) Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2). Nature 417(6885):141–147PubMedCrossRefGoogle Scholar
- Herron PR, Hughes G, Chandra G, Fielding S, Dyson PJ (2004) Transposon Express, a software application to report the identity of insertions obtained by comprehensive transposon mutagenesis of sequenced genomes: analysis of the preference for in vitro Tn5 transposition into GC-rich DNA. Nucleic Acids Res 32(14):e113PubMedCentralPubMedCrossRefGoogle Scholar
- Hobbs G, Frazer CM, Gardner DCJ, Flett F, Oliver SG (1990) Pigmented antibiotic production by Streptomyces coelicolor A3(2): kinetics and the influence of nutrients. Microbiology 136:2291–2296Google Scholar
- Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics, 2000th edn. Norwich, UKGoogle Scholar
- Molina I, Pellicer MT, Badia J, Aguilar J, Baldoma L (1994) Molecular characterization of Escherichia coli malate synthase G. Differentiation with the malate synthase A isoenzyme. FEBS J 224(2):541–548Google Scholar
- Nygaard P (1983) Utilization of preformed purine bases and nucleosides. In: Munch-Petersen A (ed) Metabolism of nucleotides, nucleosides and nucleobases in microorganisms. Academic, London, UK, pp 27–93Google Scholar
- Ramazzina I, Cendron L, Folli C, Berni R, Monteverdi D, Zanotti G, Percudani R (2008) Logical identification of an allantoinase analog (puuE) recruited from polysaccharide deacetylases. J Biol Chem 283(34):23295–23304Google Scholar
- Shilov IV, Seymour SL, Patel AA, Loboda A, Tang WH, Keating SP, Hunter CL, Nuwaysir LM, Schaeffer DA (2007) The Paragon Algorithm, a next generation search engine that uses sequence temperature values and feature probabilities to identify peptides from tandem mass spectra. Mol Cell Proteomics 6(9):1638–1655PubMedCrossRefGoogle Scholar