Large-scale production and purification of recombinant Galanthus nivalis agglutinin (GNA) expressed in the methylotrophic yeast Pichia pastoris
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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.
- 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.
- 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.
- 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.
- 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.
- 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. Phytochemistry (Oxf.) 30: 509–514.
- 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.
- Large-scale production and purification of recombinant Galanthus nivalis agglutinin (GNA) expressed in the methylotrophic yeast Pichia pastoris
Volume 25, Issue 15 , pp 1281-1285
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers-Plenum Publishers
- Additional Links
- fed-batch culture
- Galanthus nivalis agglutinin
- large-scale purification
- Pichia pastoris
- Industry Sectors
- Author Affiliations
- 1. Quality, Health & Nutrition Group, Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, Scotland
- 2. Department Agricultural and Environmental Sciences, King George VI Building, University of Newcastle, Newcastle upon Tyne, NE1 7RU, UK
- 3. Unité de Biotechnologie, Institut Meurice, Bat. 2 Campus CERIA, Av. E Gryzon, 1070, Brussels, Belgium