Biotechnology Letters

, Volume 25, Issue 15, pp 1281–1285 | Cite as

Large-scale production and purification of recombinant Galanthus nivalis agglutinin (GNA) expressed in the methylotrophic yeast Pichia pastoris

  • Philippe Baumgartner
  • Karen Harper
  • Romaan J.M. Raemaekers
  • Alain Durieux
  • Angharad M.R. Gatehouse
  • Howard V. Davies
  • Mark A. Taylor
Article

Abstract

The gene coding for agglutinin from Galanthus nivalis (GNA) was expressed in, and secreted by, the methylotrophic yeast, Pichia pastoris. Transformants of P. pastoris were selected and a process to produce and purify gram quantities of recombinant GNA was developed. GNA was secreted at approximately 80 mg l−1 at the 200 l scale and was purified to 95% homogeneity using hydrophobic interaction chromatography. The recombinant protein was similar to the protein synthesised in plant with respect to structure and biological activity.

fed-batch culture Galanthus nivalis agglutinin large-scale purification lectin Pichia pastoris 

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References

  1. Baumgartner P, Raemaekers RJM, Durieux A, Gatehouse A, Davies H, Taylor M (2002) Large-scale production, purification and characterisation of recombinant Phaeseolus vulgaris phytohemagglutinin E-form expressed in the methylotrophic yeast Pichia pastoris. Prot. Expr. Purif. 26: 394–405.Google Scholar
  2. Holmquist M, Tessier DC, Cygler M (1997) High-level production of recombinant Geotrichum candidum lipases in the yeast Pichia pastoris. Prot. Expr. Purif. 11: 35–40.Google Scholar
  3. Longstaff M, Powell KS, Gatehouse JA, Raemaekers R, Nevell CA, Hamilton WDO (1998) Production and purification of active snowdrop lectin in Escherichia coli. Eur. J. Biochem. 252: 59–65.Google Scholar
  4. Raemaekers RJM, de Muro L, Gatehouse JA, Fordham-Skelton AP (1999) Functional phytohemagglutinin (PHA) and Galanthus nivalis agglutinin (GNA) expressed in Pichia pastoris. Eur. J. Biochem. 265: 394–403.Google Scholar
  5. Shibuya N, Goldstein IJ, Van Damme EJM, Peumans WJ (1988) Binding properties of a mannose-specific lectin from the snowdrop (Galanthus nivalis) bulb. J. Biol. Chem. 263: 728–734.Google Scholar
  6. Van Damme EJM, Goldstein IJ, Peumans WJ (1991) A comparative study of mannose-binding lectins from the Amaryllidaceae and Alliaceae. Phytochemistry (Oxf.) 30: 509–514.Google Scholar
  7. Van Damme EJM, Peumans W, Pustzai A, Bardocz S (1998) Handbook of Plant Lectins: Properties and Biomedical Applications Chichester, UK: John Wiley and Sons Ltd.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Philippe Baumgartner
    • 1
  • Karen Harper
    • 1
  • Romaan J.M. Raemaekers
    • 2
  • Alain Durieux
    • 3
  • Angharad M.R. Gatehouse
    • 2
  • Howard V. Davies
    • 1
  • Mark A. Taylor
    • 1
  1. 1.Quality, Health & Nutrition GroupScottish Crop Research InstituteDundeeScotland
  2. 2.Department Agricultural and Environmental Sciences, King George VI BuildingUniversity of NewcastleNewcastle upon TyneUK
  3. 3.Unité de BiotechnologieInstitut MeuriceBrusselsBelgium

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