Applied Microbiology and Biotechnology

, Volume 97, Issue 6, pp 2493–2502 | Cite as

Exploration of two epimerase homologs in Streptomyces peucetius ATCC 27952

  • Bijay Singh
  • Tae Jin Oh
  • Jae Kyung Sohng
Applied Genetics and Molecular Biotechnology


Streptomyces peucetius ATCC 27952 is a potent producer of the therapeutically important antitumor drug, doxorubicin. S. peucetius contains two deoxythymidine diphospho (dTDP)-4-keto-6-deoxyglucose 3,5-epimerase-encoding genes, dnmU and rmbC, in its genome. While dnmU from the doxorubicin biosynthesis gene cluster is involved in the biosynthesis of dTDP-l-daunosamine, rmbC is involved in the biosynthesis of dTDP-l-rhamnose, a precursor of cell wall biosynthesis. The proteins encoded by dnmU and rmbC share 47 % identity and 64 % similarity with each other. Both enzymes converted the same substrate, dTDP-4-keto-6-deoxy-d-glucose, into dTDP-4-keto-l-rhamnose in vitro. However, when disruption of dnmU or rmbC was carried out, neither gene in S. peucetius compensated for each other’s loss of function in vivo. These results demonstrated that although dnmU and rmbC encode for similar functional proteins, their native roles in their respective biosynthetic pathways in vivo are specific and independent of one other. Moreover, the disruption of rmbC resulted in fragmented mycelia that quickly converted into gray pigmented spores. Additionally, the production of doxorubicin, a major product of S. peucetius, appeared to be abolished after the disruption of rmbC, demonstrating its pleiotropic effect. This adverse effect might have switched on the genes encoding for spore formation, arresting the expression of many genes and, thereby, preventing the production of other metabolites.


Streptomyces peucetius dTDP-4-keto-6-deoxyglucose 3,5-epimerase gene Sporulation Doxorubicin 



This work was supported by a grant from the Next-Generation BioGreen 21 Program (SSAC, grant no. PJ008013), Rural Development Administration, and the Converging Research Center Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (20090082333), Republic of Korea.

Supplementary material

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  1. Alberts B, Bray D, Hopkin K, Johnson A, Lewis J, Raff M, Roberts K, Walter P (2009) Essential cell biology. Chapter 8: control of gene expression, 3rd edn. Garland Science, New York, pp 269–294Google Scholar
  2. Bierman M, Logan R, O’Brien K, Seno ET, Rao RN, Schoner BE (1992) Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43–49CrossRefGoogle Scholar
  3. Christendat D, Saridakis V, Dharamsi A, Bochkarev A, Pai EF, Arrowsmith CH, Edwards AM (2000) Crystal structure of dTDP-4-keto-6-deoxy-d-hexulose 3,5-epimerase from Methanobacterium thermoautotrophicum complexed with dTDP. J Biol Chem 275:24608–24612CrossRefGoogle Scholar
  4. Dong C, Major LL, Srikannathasan V, Errey JC, Giraud MF, Lam JS, Graninger M, Messner P, McNeil MR, Field RA, Whitfield C, Naismith JH (2007) RmlC, a C3′ and C5′ carbohydrate epimerase, appears to operate via an intermediate with an unusual twist boat conformation. J Mol Biol 365:146–159CrossRefGoogle Scholar
  5. Driks A (2002) Overview: development in bacteria: spore formation in Bacillus subtilis. Cell Mol Life Sci 59:389–391CrossRefGoogle Scholar
  6. Dunwell JM, Culham A, Carter CE, Sosa-Aguirre CR, Goodenough PW (2001) Evolution of functional diversity in the cupin superfamily. Trends Biochem Sci 26:740–746CrossRefGoogle Scholar
  7. Dunwell JM, Purvis A, Khuri S (2004) Cupins: the most functionally diverse protein superfamily? Phytochemistry 65:7–17CrossRefGoogle Scholar
  8. Fisher SH, Sonenshein AL (1991) Control of carbon and nitrogen metabolism in Bacillus subtilis. Annu Rev Microbiol 45:107–135CrossRefGoogle Scholar
  9. Gallo MA, Ward J, Hutchinson CR (1996) The dnrM gene in Streptomyces peucetius contains a naturally occurring frameshift mutation that is suppressed by another locus outside of the daunorubicin-production gene cluster. Microbiology 142:269–275CrossRefGoogle Scholar
  10. Giraud MF, Leonard GA, Field RA, Berlind C, Naismith JH (2000) RmlC, the third enzyme of dTDP-l-rhamnose pathway, is a new class of epimerase. Nat Struct Mol Biol 7:398–402CrossRefGoogle Scholar
  11. Hutchinson CR, Colombo AL (1999) Genetic engineering of doxorubicin production in Streptomyces peucetius: a review. J Ind Microbiol Biotechnol 23:647–652CrossRefGoogle Scholar
  12. Kato J, Fujisaki S, Nakajima K, Nishimura Y, Sato M, Nakano A (1999) The Escherichia coli homologue of yeast RER2, a key enzyme of dolichol synthesis, is essential for carrier lipid formation in bacterial cell wall synthesis. J Bacteriol 181:2733–2738Google Scholar
  13. Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces Genetics. John Innes Foundation, NorwichGoogle Scholar
  14. Kornberg A, Spudich JA, Nelson DL, Deutscher MP (1968) Origin of proteins in sporulation. Annu Rev Biochem 37:51–78CrossRefGoogle Scholar
  15. Liu HW, Thorson JS (1994) Pathways and mechanisms in the biogenesis of novel deoxysugars by bacteria. Annu Rev Microbiol 48:223–256CrossRefGoogle Scholar
  16. MacNeil DJ, Occi JL, Gewain KM, MacNeil T, Gibbons PH, Ruby CL, Danis SJ (1992) Complex organization of the Streptomyces avermitilis genes encoding the avermectin polyketide synthase. Gene 115:119–125CrossRefGoogle Scholar
  17. Madduri K, Waldron C, Merlo DJ (2001) Rhamnose biosynthesis pathway supplies precursors for primary and secondary metabolism in Saccharopolyspora spinosa. J Bacteriol 183:5632–5638CrossRefGoogle Scholar
  18. Olano C, Lomovskaya N, Fonstein L, Roll JT, Hutchinson CR (1999) A two-plasmid system for the glycosylation of polyketide antibiotics: bioconversion of epsilon-rhodomycinone to rhodomycin D. Chem Biol 6:845–855CrossRefGoogle Scholar
  19. Otten SL, Gallo MA, Madduri K, Liu X, Hutchinson CR (1997) Cloning and characterization of the Streptomyces peucetius dnmZUV genes encoding three enzymes required for biosynthesis of the daunorubicin precursor thymidine diphospho-l-daunosamine. J Bacteriol 179:4446–4450Google Scholar
  20. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
  21. Singh B, Oh TJ, Sohng JK (2009) Exploration of geosmin synthase from Streptomyces peucetius ATCC 27952 by deletion of doxorubicin biosynthetic gene cluster. J Ind Microbiol Biotechnol 36:1257–1265CrossRefGoogle Scholar
  22. Singh B, Lee CB, Sohng JK (2010) Precursor for biosynthesis of sugar moiety of doxorubicin depends on rhamnose biosynthetic pathway in Streptomyces peucetius ATCC 27952. Appl Microbiol Biotechnol 85:1565–1574CrossRefGoogle Scholar
  23. Singh B, Lee CB, Park JW, Sohng JK (2012) The amino acid sequences in the C-terminal region of glucose-1-phosphate thymidylyltransferases determine their soluble expression in Escherichia coli. Protein Eng Des Sel 25:179–187CrossRefGoogle Scholar
  24. Stockmann M, Piepersberg W (1992) Gene probes for the detection of 6-deoxyhexose metabolism in secondary metabolite-producing streptomycetes. FEMS Microbiol Lett 69:185–189CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Department of Pharmaceutical Engineering, Institute of Biomolecule ReconstructionSunMoon UniversityAsansiRepublic of Korea
  2. 2.Research Institute for Bioscience & BiotechnologyKathmanduNepal

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