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Expression of Intracellular Hemoglobin Improves Protein Synthesis in Oxygen-Limited Escherichia coli

Bio/Technology volume 8, pages 849–853 (1990)Cite this article

Abstract

We have previously cloned the Vitreoscilla hemoglobin gene (VHb) and expressed the protein in Escherichia coli in its active form. Under oxygen-limited conditions the presence of VHb improves protein synthesis as indicated by both total protein content and the activity of an enzyme expressed from a cloned gene present on a multicopy plasmid. Measurements of nitrogen utilization rates corroborate the observation of enhanced protein synthesis; however, the rates of carbon consumption and acid synthesis remain unchanged. This suggests that the net effect of VHb in E. coli is to improve the efficiency, rather than the kinetics, of oxygen-limited aerobic metabolism. We propose two possible models for the mechanism of action of VHb: the facilitated diffusion hypothesis and the intracellular redox effector hypothesis. These suggest other systems in which cloned VHb may enhance bioprocess productivity.

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References

  1. Poole, R.K. and Ingledew, W.J. 1987. Pathways of electrons to oxygen, p. 170–200. In: Escherichia coli and Salmonella typhimurium. Neidhardt, F. C. (Ed. in chief). American Society for Microbiology, Washington, D.C.

    Google Scholar 

  2. Anraku, Y. and Gennis, R.B. 1987. The aerobic respiratory chain of Escherichia coli. Trends Biochem. Sci. 12: 262–266.

    Article  CAS  Google Scholar 

  3. Anraku, Y. 1988. Bacterial electron transport chains. Ann. Rev. Biochem. 57: 101–132.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  6. Dikshit, K.L. and Webster, D.A. 1988. Cloning, characterization and expression of the bacterial globin gene from Vitreoscilla in Escherichia coli. Gene 70: 377–386.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  8. Khosla, C. and Bailey, J.E. 1989. Evidence for partial export of Vitreoscilla hemoglobin into the periplasmic space in Escherichia coli: Implications for protein function. J. Mol. Biol. 210: 79–90.

    Article  CAS  PubMed  Google Scholar 

  9. Khosla, C. and Bailey, J.E. 1989. Characterization of the oxygen-dependent promoter of the Vitreoscilla hemoglobin gene in Escherichia coli. J. Bacteriol. 171: 5995–6004.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Miller, J.H. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

  11. Georgiou, C.D. and Webster, D.A. 1987. Identification of b, c, and d cytochromes in the membrane of Vitreoscilla. Arch. Microbiol. 148: 328–333.

    Article  CAS  PubMed  Google Scholar 

  12. Tempest, D.W. and Neijssel, O.M. 1987. Growth yield and energy distribution, p. 797–806. In: Escherichia coli and Salmonella typhimurium. Neidhardt, F. C. (Ed. in chief). American Society for Microbiology, Washington, D.C.

    Google Scholar 

  13. Doelle, H.W., Ewings, K.N. and Hollywood, N.W. 1983. Regulation of glucose metabolism in bacterial systems. Adv. Biochem. Eng. 23: 1–31.

    Google Scholar 

  14. Wittenberg, J. 1966. The molecular mechanism of hemoglobin-facilitated oxygen diffusion. J. Biol. Chem. 241: 104–114.

    CAS  PubMed  Google Scholar 

  15. 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  PubMed  Google Scholar 

  16. Hughes, D.E., Curtis, J.E., Khosla, C. and Bailey, J.E. 1989. A new oxygen-regulated promoter for the expression of proteins in Escherichia coli. Biotechniques 7: 1026–1028.

    CAS  PubMed  Google Scholar 

  17. Weisemann, J.M. and Weinstock, G.E. 1985. Direct selection of mutations reducing transcription or translation of the recA gene of Escherichia coli with a recA-lacZ protein fusion. J. Bacteriol. 163: 748–755.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Neumann, J.R., Morency, C.A. and Russian, K.O. 1987. A novel rapid assay for chloramphenicol acetyltransferase gene expression. Biotechniques 5: 444–447.

    CAS  Google Scholar 

  19. Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  CAS  PubMed  Google Scholar 

  20. O'Farrell, P.H. 1975. High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250: 4007–4021.

    CAS  PubMed  Google Scholar 

  21. Hochstrasser, D.F., Harrington, M.G., Hochstrasser, A.C., Miller, M.J. and Merril, C.R. 1988. Methods for increasing the resolution of two-dimensional protein electrophoresis. Anal. Biochem. 173: 424–435.

    Article  CAS  PubMed  Google Scholar 

  22. Phillips, T.A., Vaughn, V., Bloch, P.L. and Neidhardt, F.C. 1987. Gene-protein index of Escherichia coli K12, edition 2, p. 919–966. In: Escherichia coli and Salmonella typhimurium. Neidhardt, F. C. (Ed. in chief). American Society for Microbiology, Washington, D.C.

    Google Scholar 

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Author notes

  1. Chaitan Khosla

    Present address: Department of Genetics, John Innes Institute, Norwich, NR4 7UH, U.K.

  2. James E. Bailey: Corresponding author.

Authors and Affiliations

  1. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, 91125

    Ursula Rinas & James E. Bailey

  2. Exogene, Pasadena, CA, 91107

    Joseph E. Curtis & John DeModena

Authors
  1. Chaitan Khosla
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  2. Joseph E. Curtis
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  3. John DeModena
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  4. Ursula Rinas
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  5. James E. Bailey
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Khosla, C., Curtis, J., DeModena, J. et al. Expression of Intracellular Hemoglobin Improves Protein Synthesis in Oxygen-Limited Escherichia coli. Nat Biotechnol 8, 849–853 (1990). https://doi.org/10.1038/nbt0990-849

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