Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Jun;178(11):3384–3388. doi: 10.1128/jb.178.11.3384-3388.1996

Cloning, sequencing, and analysis of aklaviketone reductase from Streptomyces sp. strain C5.

M L Dickens 1, J Ye 1, W R Strohl 1
PMCID: PMC178101  PMID: 8655529

Abstract

DNA sequence analysis of a region of the Streptomyces sp. strain C5 daunomycin biosynthesis gene cluster, located just upstream of the daunomycin polyketide biosynthesis genes, revealed the presence of six complete genes. The two genes reading right to left include genes encoding the potentially translationally coupled gene products, an acyl carrier protein and a ketoreductase, and the four genes reading divergently, left to right, include two open reading frames of unknown function followed by a gene encoding an apparent glycosyltransferase and dauE, encoding aklaviketone reductase. Extracts of Streptomyces lividans TK24 containing recombinant DauE catalyzed the NADPH-specific conversion of aklaviketone, maggiemycin, and 7-oxodaunomycinone to aklavinone, epsilon-rhodomycinone, and daunomycinone, respectively. Neither the product of dauB nor that of the ketoreductase gene directly downstream of the acyl carrier protein gene demonstrated aklaviketone reductase activity.

Full Text

The Full Text of this article is available as a PDF (297.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bibb M. J., Findlay P. R., Johnson M. W. The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences. Gene. 1984 Oct;30(1-3):157–166. doi: 10.1016/0378-1119(84)90116-1. [DOI] [PubMed] [Google Scholar]
  3. Bibb M. J., Sherman D. H., Omura S., Hopwood D. A. Cloning, sequencing and deduced functions of a cluster of Streptomyces genes probably encoding biosynthesis of the polyketide antibiotic frenolicin. Gene. 1994 May 3;142(1):31–39. doi: 10.1016/0378-1119(94)90351-4. [DOI] [PubMed] [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dickens M. L., Strohl W. R. Isolation and characterization of a gene from Streptomyces sp. strain C5 that confers the ability to convert daunomycin to doxorubicin on Streptomyces lividans TK24. J Bacteriol. 1996 Jun;178(11):3389–3395. doi: 10.1128/jb.178.11.3389-3395.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dickens M. L., Ye J., Strohl W. R. Analysis of clustered genes encoding both early and late steps in daunomycin biosynthesis by Streptomyces sp. strain C5. J Bacteriol. 1995 Feb;177(3):536–543. doi: 10.1128/jb.177.3.536-543.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eckardt K., Schumann G., Tresselt D., Ihn W. Biosynthesis of anthracyclinones: isolation of a new early cyclization product aklaviketone. J Antibiot (Tokyo) 1988 Jun;41(6):788–793. doi: 10.7164/antibiotics.41.788. [DOI] [PubMed] [Google Scholar]
  8. Eckardt K., Wagner C. Biosynthesis of anthracyclinones. J Basic Microbiol. 1988;28(1-2):137–144. doi: 10.1002/jobm.3620280117. [DOI] [PubMed] [Google Scholar]
  9. Fernández-Moreno M. A., Martínez E., Caballero J. L., Ichinose K., Hopwood D. A., Malpartida F. DNA sequence and functions of the actVI region of the actinorhodin biosynthetic gene cluster of Streptomyces coelicolor A3(2). J Biol Chem. 1994 Oct 7;269(40):24854–24863. [PubMed] [Google Scholar]
  10. Grimm A., Madduri K., Ali A., Hutchinson C. R. Characterization of the Streptomyces peucetius ATCC 29050 genes encoding doxorubicin polyketide synthase. Gene. 1994 Dec 30;151(1-2):1–10. doi: 10.1016/0378-1119(94)90625-4. [DOI] [PubMed] [Google Scholar]
  11. Hallam S. E., Malpartida F., Hopwood D. A. Nucleotide sequence, transcription and deduced function of a gene involved in polyketide antibiotic synthesis in Streptomyces coelicolor. Gene. 1988 Dec 30;74(2):305–320. doi: 10.1016/0378-1119(88)90165-5. [DOI] [PubMed] [Google Scholar]
  12. Hopwood D. A., Sherman D. H. Molecular genetics of polyketides and its comparison to fatty acid biosynthesis. Annu Rev Genet. 1990;24:37–66. doi: 10.1146/annurev.ge.24.120190.000345. [DOI] [PubMed] [Google Scholar]
  13. Lampel J. S., Aphale J. S., Lampel K. A., Strohl W. R. Cloning and sequencing of a gene encoding a novel extracellular neutral proteinase from Streptomyces sp. strain C5 and expression of the gene in Streptomyces lividans 1326. J Bacteriol. 1992 May;174(9):2797–2808. doi: 10.1128/jb.174.9.2797-2808.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Le Gouill C., Desmarais D., Déry C. V. Saccharopolyspora hirsuta 367 encodes clustered genes similar to ketoacyl synthase, ketoacyl reductase, acyl carrier protein, and biotin carboxyl carrier protein. Mol Gen Genet. 1993 Jul;240(1):146–150. doi: 10.1007/BF00276894. [DOI] [PubMed] [Google Scholar]
  15. Madduri K., Hutchinson C. R. Functional characterization and transcriptional analysis of a gene cluster governing early and late steps in daunorubicin biosynthesis in Streptomyces peucetius. J Bacteriol. 1995 Jul;177(13):3879–3884. doi: 10.1128/jb.177.13.3879-3884.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Otten S. L., Liu X., Ferguson J., Hutchinson C. R. Cloning and characterization of the Streptomyces peucetius dnrQS genes encoding a daunosamine biosynthesis enzyme and a glycosyl transferase involved in daunorubicin biosynthesis. J Bacteriol. 1995 Nov;177(22):6688–6692. doi: 10.1128/jb.177.22.6688-6692.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pandey R. C., Toussaint M. W., McGuire J. C., Thomas M. C. Maggiemycin and anhydromaggiemycin: two novel anthracyclinone antitumor antibiotics. Isolation, structures, partial synthesis and biological properties. J Antibiot (Tokyo) 1989 Nov;42(11):1567–1577. doi: 10.7164/antibiotics.42.1567. [DOI] [PubMed] [Google Scholar]
  18. Sherman D. H., Malpartida F., Bibb M. J., Kieser H. M., Bibb M. J., Hopwood D. A. Structure and deduced function of the granaticin-producing polyketide synthase gene cluster of Streptomyces violaceoruber Tü22. EMBO J. 1989 Sep;8(9):2717–2725. doi: 10.1002/j.1460-2075.1989.tb08413.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Strohl W. R., Connors N. C. Significance of anthraquinone formation resulting from the cloning of actinorhodin genes in heterologous streptomycetes. Mol Microbiol. 1992 Jan;6(2):147–152. doi: 10.1111/j.1365-2958.1992.tb01995.x. [DOI] [PubMed] [Google Scholar]
  20. Tsukamoto N., Fujii I., Ebizuka Y., Sankawa U. Nucleotide sequence of the aknA region of the aklavinone biosynthetic gene cluster of Streptomyces galilaeus. J Bacteriol. 1994 Apr;176(8):2473–2475. doi: 10.1128/jb.176.8.2473-2475.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Vara J., Lewandowska-Skarbek M., Wang Y. G., Donadio S., Hutchinson C. R. Cloning of genes governing the deoxysugar portion of the erythromycin biosynthesis pathway in Saccharopolyspora erythraea (Streptomyces erythreus). J Bacteriol. 1989 Nov;171(11):5872–5881. doi: 10.1128/jb.171.11.5872-5881.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wright F., Bibb M. J. Codon usage in the G+C-rich Streptomyces genome. Gene. 1992 Apr 1;113(1):55–65. doi: 10.1016/0378-1119(92)90669-g. [DOI] [PubMed] [Google Scholar]
  23. Ye J., Dickens M. L., Plater R., Li Y., Lawrence J., Strohl W. R. Isolation and sequence analysis of polyketide synthase genes from the daunomycin-producing Streptomyces sp. strain C5. J Bacteriol. 1994 Oct;176(20):6270–6280. doi: 10.1128/jb.176.20.6270-6280.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yu T. W., Chen C. W. The unstable melC operon of Streptomyces antibioticus is codeleted with a Tn4811-homologous locus. J Bacteriol. 1993 Mar;175(6):1847–1852. doi: 10.1128/jb.175.6.1847-1852.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES