Skip to main content
SpringerLink
Log in

Expression of Bacterial Hemoglobin in the Yeast, Pichia pastoris, with a Low O2-Induced Promoter

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

The PsADH2-promoter of Pichia stipitis alcohol dehydrogenase II (ADH II) gene was employed to control the expression of Vitreoscilla hemoglobin (VHb) gene in Pichia pastoris. As in P. stipitis, the promoter was also induced microaerobically in P. pastoris. The expression level of VHb in P. pastoris at low O2 tension (<5% air saturation) was 16 nmol/g dry cell wt, i.e. about 24-fold higher than that at 60% air saturation. The expressed VHb enhanced growth of P. pastoris under microaerobic conditions. The application of O2-regulated promoter in P. pastoris revealed that induction of high-level expression of heterologous protein is feasible without addition of supplementary compounds.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • SL Bhave BB Chattoo (2003) ArticleTitleExpression of Vitreoscilla hemoglobin improves growth and levels of extracellular enzyme in Yarrowia lipolytica Biotechnol. Bioeng. 84 658–666

    Google Scholar 

  • L Bulow N Holmberg G Lilius E Bailey (1999) ArticleTitleThe metabolic effects of native and transgenic hemoglobins on plants Trends Biotechnol. 17 21–24

    Google Scholar 

  • W Chen DE Hughes JE Bailey (1994) ArticleTitleIntracellular expression of Vitreoscilla hemoglobin alters aerobic metabolism of Saccharomyces cerevisiae Biotechnol. Progr. 10 308–313

    Google Scholar 

  • JM Cregg TS Vedvick WC Raschke (1993) ArticleTitleRecent advances in the expression of foreign genes in Pichia pastoris BioTechnology 11 905–910

    Google Scholar 

  • KL Dikshit DA Webster (1988) ArticleTitleCloning, characterization and expression of the bacterial hemoglobin gene from Vitreoscilla in Escherichia coli Gene 70 377–386

    Google Scholar 

  • AD Frey PT Kallio (2003) ArticleTitleBacterial hemoglobins and flavohemoglobins: versatile proteins and their impact on microbiology and biotechnology FEMS Microbiol. Rev. 27 525–545

    Google Scholar 

  • JF Gorgens V Passoth WH Zyl Particlevan JH Knoetze B Hahn-Hagerdal (2005) ArticleTitleAmino acid supplementation, controlled oxygen limitation and sequential double induction improves heterologous xylanase production by Pichia stipitis FEMS Yeast Res. 5 677–683

    Google Scholar 

  • RM Horton HD Hunt SN Ho JM Pullen LR Pease (1989) ArticleTitleEngineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension Gene 77 61–68

    Google Scholar 

  • TW Jeffries CP Kurtzman (1994) ArticleTitleStrain selection, taxonomy, and genetics of xylose-fermenting yeasts Enzyme Microb. Technol. 16 922–932

    Google Scholar 

  • CY Liu DA Webster (1974) ArticleTitleSpectral characteristics and interconversions of the reduced, oxidized, oxygenated forms of purified cytochrome o J. Biol. Chem. 249 4261–4266

    Google Scholar 

  • V Passoth B Schafer B Liebel T Weierstall U Klinner (1998) ArticleTitleMolecular cloning of alcohol dehydrogenase genes of the yeast Pichia stipitis and identification of the fermentative ADH Yeast 14 1311–1325

    Google Scholar 

  • V Passoth B Hahn-Hagerdal (2000) ArticleTitleProduction of a heterologous endo-1,4-β-xylanase in the yeast Pichia stipitis with an O2-regulated promoter Enzyme Microb. Technol. 26 781–784

    Google Scholar 

  • V Passoth M Cohn B Schafer B Hahn-Hagerdal U Klinner (2003) ArticleTitleAnalysis of the hypoxia-induced ADH2 promoter of the respiratory yeast Pichia stipitis reveals a new mechanism for sensing of oxygen limitation in yeast Yeast 20 39–51

    Google Scholar 

  • S Shen G Sulter TW Jeffries JM Cregg (1998) ArticleTitleA strong nitrogen source-regulated promoter for controlled expression of foreign genes in the yeast Pichia pastoris Gene 216 93–102

    Google Scholar 

  • JF Tschopp PF Brust JM Cregg CA Stillman TR Gingeras (1987) ArticleTitleExpression of the lacZ gene from two methanol-regulated promoters in Pichia pastoris Nucleic Acids Res. 15 3859–3876

    Google Scholar 

  • HR Waterham ME Digan PJ Koutz SV Lair JM Cregg (1997) ArticleTitleIsolation of the Pichia pastoris glyceraldehydes-s-phosphate dehydrogenase gene and regulation and use of its promoter Gene 186 37–44

    Google Scholar 

  • S Wu GJ Letchworth (2004) ArticleTitleHigh efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol Biotechniques 36 152–154

    Google Scholar 

  • JM Wu TA Hsu CK Lee (2003) ArticleTitleExpression of the gene coding for bacterial hemoglobin improves β-galactosidase production in a recombinant Pichia pastoris Biotechnol. Lett. 25 1457–1462

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemical Engineering, National Taiwan University of Science and Technology, 106, Taipei, Taiwan

    Liang-Jung Chien & Cheng-Kang Lee

Authors
  1. Liang-Jung Chien
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cheng-Kang Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cheng-Kang Lee.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chien, LJ., Lee, CK. Expression of Bacterial Hemoglobin in the Yeast, Pichia pastoris, with a Low O2-Induced Promoter. Biotechnol Lett 27, 1491–1497 (2005). https://doi.org/10.1007/s10529-005-1324-x

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-005-1324-x

Keywords

Search

Navigation