유사 생합성 경로를 가진 Streptomyces sp.의 혼합배양을 이용한 Doxorubicin 생합성

  • Choi, Yun-Hwa (Department of Microbiology, Chungbuk National University) ;
  • Hong, Young-Soo (Korea Research Institute of Bioscience and Biotechnology) ;
  • Lim, Jai-Yun (Department of Microbiology, Chungbuk National University) ;
  • Lee, Jung-Joon (Korea Research Institute of Bioscience and Biotechnology)
  • 최윤화 (충북대학교 자연과학대학 미생물학과) ;
  • 홍영수 (생명공학연구소 천연물생합성 R.U.) ;
  • 임재윤 (충북대학교 자연과학대학 미생물학과) ;
  • 이정준 (생명공학연구소 천연물생합성 R.U.)
  • Published : 1997.12.01

Abstract

We selected two mutants namely strain D5 and Nu23 by mutagenesis of anthracycline producing Streptomyces: the former is an $\varepsilon$-rhodomycinone overproducing mutant selected from Streptomyces sp. C5, a baumycin producer and the latter, a blocked mutant of early pathway for doxorubicin biosynthesis obtained from Streptomyces peucetius ATCC 27952, a doxorubicin producer. The mutant strain Nu23 does not produce anthracycline metabolites but retains the most of enzyme activities converting aklavinone to doxorubicin and the mutant strain D5 produced $\varepsilon$-rhodomycinone at a level of 150 $\mu$g/ml. These strains were grown separately in NDYE medium and each was mixed at day 3 by equal volume of culture broth but the quantity of doxorubicin produced was far below an estimation based on the level of $\varepsilon$-rhodomycinone normaly produced by the strain D5. On the other hand doxorubicin was reached at maximum level after 4 days in the mixed culture condition which was composed of culture broth of strain D5 grown for 6 day and that of strain Nu23 grown for 3 day. It was turned out that the growth of mutant strain D5 was inhibited by the accumulation of daunorubicin and doxorubicin in mixed culture broth, which cause the limitation of $\varepsilon$-rhodomycinone.

Keywords

Streptomyces;Mutant;Mixed culture;Doxorubicin

References

  1. Nature v.314 Production of hybrid antibiotics by genetic engineering Hopwood,D.A.;F.Malpartida;H.M.Kieser;H.Ikeda;J.Duncan;I.Fujii;B.A.Rudd;H.G.Floss;S.Omura
  2. Biotechnology of industrial antibiotics v.22 Daunorubicin and adriamycin:properties, biosynthesis and fermentation White,R.J.;R.M.Stroshane;E.J.Vandamme(ed.)
  3. J. Microbiol. Biotechnol. v.7 Characterization of doxorubicinnonproducing mutant, Nu23 of Streptomyces peucetius ATCC 27952 Hwang,C.K.;H.S.Lee;Y.S.Hong;N.S.Paek;T.H.Kim;J.J.Lee
  4. Can. J. Microbiol. v.31 Nutrient effects on anthracycline production by Streptomyces peucetius in a defined medium Dekleva,M.L.;J.A.Titus;W.R.Strohl
  5. J. Bacteriol. v.172 Cloning and expression of daunorubicin biosynthesis genes from Streptomyces peucetius and Streptomyces peucetius subsp. caesius. Otten,S.L.;K.J.Stutzman Engwall;C.R.Hutchinson
  6. Cancer Treat. Rep. v.63 Daunorubicin and adriamycin in cancer treatment: an analysis of their roles and limitations Davis,H.L.;T.E.Davis
  7. J. Gen. Microbiol. v.136 Biosynthesis of anthracyclines: analysis of mutants of Streptomyces sp. strain C5 blocked in daunomycin biosynthesis Bartel,P.L.;N.C.Connors;W.R.Strohl
  8. J. Fermen. Bioengin v.79 Daunomycin biosynthesis by microbial conversion of precursor metabolites using biosynthetically blocked mutants Yoshimoto,A.;T.Nakamura;K.Kubo;O.Johdo;H.Tone
  9. Nature v.375 Rational design of aromatic polyketide natural products by recombinant assembly of enzymatic subunits McDaniel,R.;S.Ebert Khosla;D.A.Hopwood;C.Khosla
  10. Biotechnol. Bioeng. v.11 Adriamycin, 14-Hydroxydaunomycin, a new antitumor antibiotic from Streptomyces peucetius var caesius Arcamone,F.;G.Cassinelli;G.Fantini;A.Grein;P.Orezzi;C.Pol;C.Spalla
  11. Proc. Natl. Acad. Sci. v.84 Cloning genes for the biosynthesis of a macrolide antibiotic Fishman,S.E.;K.Cox;J.L.Larson;P.A.Reynolds;E.T.Seno;W.K.Yen;R.Van Frank;C.L.Hershberger
  12. Genetic manipulation of Streptomyces;A laboratory manual Hopwood,D.W.;M.J.Bibb;K.F.Chater;T.Kieser;C.J.Bruton;H.M.Kieser;D.J.Lydiate;C.P.Smith;J.M.Ward;H.Schremp
  13. Proc. Natl. Acad. Sci. v.90 An erythromycin analog produced by reprogramming of polyketide synthesis Donadio,S.;J.B.Mcalpine;P.J.Sheldon;M.Jackson;L.Katz
  14. J. Bacteriol. v.178 Isolation and Characterization of a gene from Streptomyces sp. strain C5 that confers the ability to convert daunomycin to doxorubicin on Streptomyces lividans TK24 Dickens,M.L.;W.R.Strohl
  15. Mutation Research v.9 Mutagenesis by N-methyl-N-nitro-N-nitrosoguanidine(NTG) in Streptomyces coelicolor Delic,V.;D.A.Hopwood;E.Friend
  16. J. Bacteriol. v.176 Molecular cloning and characterization of the aklavinone 11-hydroxylase gene of Streptomyces peucetius subsp. caesius ATCC 27952 Hong,Y.S.;C.K.Hwang;S.K.Hong;Y.H.Kim;J.J.Lee
  17. J. Antibiotics v.49 New anthracycline Metabolites produced by the aklavinone 11-hydroxylase gene in Streptomyces galilaeus ATCC 31133 Kim,H.S.;Y.S.Hong;Y.H.Kim;O.J.Yoo;J.J.Lee
  18. Med. Res. Rev. v.4 Antitumor anthracycline: recent developments Arcamone,F.
  19. Antimicrob. Agents Chemother v.39 Expression of Streptomyces peucetius genes for doxorubicin resistance and aklavinone 11-hydroxylase in Streptomyces galilaeus ATCC 31133 and production of a hybrid aclacinomycin Hwang,C.K.;H.S.Kim;Y.S.Hong;Y.H.Kim;S.K.Hong;S.J.Kim;J.J.Lee