Large-scale production and purification of recombinant Galanthus nivalis agglutinin (GNA) expressed in the methylotrophic yeast Pichia pastoris
Cite this article as: Baumgartner, P., Harper, K., Raemaekers, R.J. et al. Biotechnology Letters (2003) 25: 1281. doi:10.1023/A:1025007901322 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 References
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.
Holmquist M, Tessier DC, Cygler M (1997) High-level production of recombinant
Geotrichum candidum lipases in the yeast Pichia pastoris. Prot. Expr. Purif.
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.
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.
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.
Van Damme EJM, Goldstein IJ, Peumans WJ (1991) A comparative study of mannose-binding lectins from the Amaryllidaceae and Alliaceae.
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. Copyright information
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