Log in

Development of a set of expression vectors in Hansenula polymorpha

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Four expression vectors based on formate dehydrogenase promoter (FMDp) and methanol oxidase promoter (MOXp) from Hansenula polymorpha were developed to express heterologous genes in Hansenula polymorpha. A secretion signal sequence of the mating factor-alpha from Saccharomyces cerevisiae was inserted in the secretory expression plasmids for efficient secretion. A modified green fluorescent protein (mGFP5) was used as the marker of expression for the first time in H. polymorpha NCYC495 (leu 1.1) to determine the expression ability of these plasmids. The mGFP5 thus expressed retained its biochemical and physiological properties, such as accumulation inside cells and efficient secretion into the culture media. These results indicated that the four integrative vectors are useful expression systems which could be directly applied for production of heterologous proteins of interests in H. polymorpha.

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

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (France)

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Abdel-Salam HA, El-Khamissy T, Enan GA, Hollenberg CP (2001) Expression of mouse anticreatine kinase (MAK33) monoclonal antibody in the yeast Hansenula polymorpha. Appl. Microbiol. Biotechnol. 56: 157–164.

    Google Scholar 

  • Aprikian PG, Karpychev IV, Mikhailover VM, Gracheva VD, Shchedrin AM, Beburov M, El'darov MA, Skriabin KG (1991) Expression and secretion of human growth hormone (hGH) in methylotrophic yeasts Hansenula polymorpha. Dokl. Akad. Nauk. SSSR. 321: 390–394.

    Google Scholar 

  • Brito MDP, Perdomo G, Gonzalez C (1999) A set of Hansenula polymorpha integrative vectors to construct LacZ fusions. Appl. Microbiol. Biotechnol. 53: 23–29.

    Google Scholar 

  • Chung CT, Niemela SL, Miller RH (1989) One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc. Natl. Acad. Sci. USA 86: 2172–2175.

    Google Scholar 

  • Faber KN, Haima P, Harder W, Veenhuis M, Ab G (1994) Highly-efficient electrotransformation of the yeast Hansenula polymorpha. Curr. Genet. 25: 305–310.

    Google Scholar 

  • Fellinger AJ, Verbakel JM, Veale RA, Sudbery PE, Bom IJ, Overbeeke N, Verrips CT (1991) Expression of the alpha-galactosidase from Cyamopsis tetragonoloba (guar) by Hansenula polymorpha. Yeast 7: 463–473.

    Google Scholar 

  • Gellissen G, Janowicz ZA, Merckelbach A, Piontek M, Keup P, Weydemann U, Hollenberg CP, Strasser AW (1991) Heterologous gene expression in Hansenula polymorpha: efficient secretion of glucoamylase. Biotechnology 9: 291–295.

    Google Scholar 

  • Hodgkins M, Mead D, Ballance DJ, Goodey A, Sudbery P (1993) Expression of the glucose oxidase gene from Aspergillus niger in Hansenula polymorpha and its use as a reporter gene to isolate regulatory mutations. Yeast 9: 625–635.

    Google Scholar 

  • Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57: 267–272.

    Google Scholar 

  • Janowicz ZA, Melber K, Merckelbach A, Jacobs E, Harford N, Comberbach M, Hollenberg CP (1991) Simultaneous expression of the S and L surface antigens of hepatitis B, and formation of mixed particles in the methylotrophic yeast, Hansenula polymorpha. Yeast 7: 431–443.

    Google Scholar 

  • Kang HA, Kang W, Hong WK, Kim MW, Kim JY, Sohn JH, Choi ES, Choe KB, Rhee SK (2001) Development of expression systems for the production of recombinant human serum albumin using the MOX promoter in Hansenula polymorpha DL-1. Biotechnol. Bioeng. 76: 175–185.

    Google Scholar 

  • Kato N, Pontier D, Lam E (2002) Spectral profiling for the simultaneous observation of four distinct fluorescent proteins and detection of protein-protein interaction via fluorescence resonance energy transfer in tobacco leaf nuclei. Plant Physiol. 129: 931–942.

    Google Scholar 

  • Mayer AF, Hellmuth K, Schlieker H, Lopez-Ulibarri R, Oertel S, Dahlems U, Strasser AW, van Loon AP (1999) An expression system matures: a highly efficient and cost-effective process for phytase production by recombinant strains of Hansenula polymorpha. Biotechnol. Bioeng. 63: 373–381.

    Google Scholar 

  • Merckelbach A, Godecke S, Janowicz ZA, Hollenberg CP (1993) Cloning and sequencing of the ura3 locus of the methylotrophic yeast Hansenula polymorpha and its use for the generation of a deletion by gene replacement. Appl. Microbiol. Biotechnol. 40: 361–364.

    Google Scholar 

  • Raschke WC, Neiditch BR, Hendricks M, Cregg JM (1996) Inducible expression of a heterologous protein in Hansenula polymorpha using the alcohol oxidase 1 promoter of Pichia pastoris. Gene 177: 163–167.

    Google Scholar 

  • Sarramegna V, Talmont F, Seree de Roch M, Milon A, Demange P (2002) Green fluorescent protein as a reporter of human muopioid receptor overexpression and localization in the methylotrophic yeast Pichia pastoris. J. Biotechnol. 99: 23–39.

    Google Scholar 

  • Weydemann U, Keup P, Piontek M, Strasser AW, Schweden J, Gellissen G, Janowicz ZA (1995) High-level secretion of hirudin by Hansenula polymorpha û authentic processing of three different preprohirudins. Appl. Microbiol. Biotechnol. 44: 377–385.

    Google Scholar 

  • Yu YA, Szalay AA (2002) A Renilla luciferase-Aequorea GFP (ruc-gfp) fusion gene construct permits real-time detection of promoter activation by exogenously administered mifepristone in vivo. Mol. Genet. Genomics 268: 169–178.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Bingsheng Qiu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Song, H., Li, Y., Fang, W. et al. Development of a set of expression vectors in Hansenula polymorpha . Biotechnology Letters 25, 1999–2006 (2003). https://doi.org/10.1023/B:BILE.0000004392.87179.29

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:BILE.0000004392.87179.29

Navigation