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Actinorhodin Production By Streptomyces coelicolor and Growth of Streptomyces lividans Are Improved by the Expression of a Bacterial Hemoglobin

Abstract

Secondary metabolite production by Streptomyces is often highly sensitive to oxygen supply, which can be limiting in large-scale fermentations. In an attempt to improve oxygen utilization by the cells, we expressed a heterologous bacterial hemoglobin gene in Streptomyces coelicolor and Streptomyces lividans. Hemoglobin expression was demonstrated by immunoblot analysis and carbon monoxide binding activity. In batch fermentations run under reduced aeration, the expression of hemoglobin in S. coelicolor resulted in a ten-fold increase in specific yields of the aromatic polyketide, actinorhodin. Actinorhodin yields were also much less sensitive to aeration conditions in the hemoglobin-expressing strain. In addition, hemoglobin-expressing S. lividans cells grown under reduced aeration had higher final cell densities and exhibited greater oxygen consumption rates than non-expressing cells.

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References

  1. Atkinson, B. and Mavituna, B. 1983. Biochemical Engineering and Biotechnology Handbook. Stockton Press, New York, NY.

    Google Scholar 

  2. Buckland, B. 1985. Fermentation exhaust gas analysis using mass spectrometry. Bio/Technology 3: 982–988.

    CAS  Google Scholar 

  3. Yegneswaran, P.K. and Gray, M.R. 1988. Effects of reduced oxygen on growth and antibiotic production in Streptomyces clavuligerus. Biotechnol. Lett. 10: 479–484.

    Article  CAS  Google Scholar 

  4. Adlercreutz, P. and Mattiasson, B. 1982. Oxygen supply to immobilized cells. Eur. J. Appl. Microbiol. Biotechnol. 16: 165–170.

    Article  CAS  Google Scholar 

  5. Khosla, C. and Bailey, J.E. 1988. Heterologous expression of a bacterial haemoglobin improves the growth properties of recombinant Escherichia coli. Nature 331: 633–635.

    Article  CAS  Google Scholar 

  6. Orii, Y. and Webster, D.A. 1986. Photodissociation of oxygenated cytochrome o(s) (Vitreoscilla) and kinetic studies of reassociation. J. Biol. Chem. 261: 3544–3547.

    CAS  Google Scholar 

  7. Boreman, S. and Webster, D.A. 1982. Control of heme content in Vitreoscilla by oxygen. J. Gen. Appl. Microbiol. 28 35–43.

    Article  Google Scholar 

  8. Wakabayashi, S., Matsubara, H. and Webster, D.A. 1986. Primary sequence of a dimeric bacterial hemoglobin from Vitreoscilla. Nature 322: 481–483.

    Article  CAS  Google Scholar 

  9. Khosla, C. and Bailey, J.E. 1988. The Vitreoscilla hemoglobin gene: Molecular cloning, nucleotide sequence and genetic expression in Escherichia coli. Mol. Gen. Genet. 214: 158–161.

    Article  CAS  Google Scholar 

  10. Khosla, C., Curtis, J.E., DeModena, J., Rinas, U. and Bailey, J.E. 1990. Expression of intracellular hemoglobin improves protein synthesis in oxygen-limited Escherichia coli. Bio/Technology 8: 849–853.

    CAS  PubMed  Google Scholar 

  11. Cole, S.P., Rudd, B.A.M., Hopwood, D.A., Chang, C. and Floss, H.G. 1987. Biosynthesis of the antibiotic actinorhodin: Analysis of blocked mutants of Streptomyces coelicolor. J. Antibiotics. 40: 340–347.

    Article  CAS  Google Scholar 

  12. Khosla, C. and Bailey, J.E. 1989. Evidence for partial export of Vitreoscilla hemoglobin into the periplasmic space in Escherichia coli. J. Mol. Biol. 210 79–89.

    Article  CAS  Google Scholar 

  13. Kieser, T. and Melton, R.E. 1988. Plasmid pIJ699, a multi-copy positive-selection vector for Streptomyces. Gene 65 83–91.

    Article  CAS  Google Scholar 

  14. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  15. Hobbs, G., Frazer, C.M., Gardner, C.J., Cullum, J.A. and Oliver, S.G. 1989. Dispersed growth of Streptomyces in liquid culture. Appl. Microbiol. Biotechnol. 31: 272–277.

    Article  CAS  Google Scholar 

  16. Hopwood, D.A., Bibb, M.J., Chater, K.F., Kieser, T., Bruton, C.J., Kieser, H.M., Lydiate, D.J., Smith, C.P., Ward, J.M. and Schrempf, H. 1985. Genetic Manipulation of Streptomyces: A Laboratory Manual. John Innes Foundation, Norwich, UK.

    Google Scholar 

  17. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. 1987. Current Protocols in Molecular Biology. Current Protocols, U.S.A.

    Google Scholar 

  18. Webster, D.A. and Lu, C.Y. 1974. Reduced nicotinamide adenine dinucleotide cytochrome o reductase associated with cytochrome o purified from Vitreoscilla. J. Biol. Chem. 249: 4257–4260.

    CAS  PubMed  Google Scholar 

  19. Bradford, M.M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principal of protein dye binding. Anal. Biochem. 72: 248–254.

    Article  CAS  Google Scholar 

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Magnolo, S., Leenutaphong, D., DeModena, J. et al. Actinorhodin Production By Streptomyces coelicolor and Growth of Streptomyces lividans Are Improved by the Expression of a Bacterial Hemoglobin. Nat Biotechnol 9, 473–476 (1991). https://doi.org/10.1038/nbt0591-473

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