Article

Biotechnology Letters

, Volume 25, Issue 15, pp 1281-1285

First online:

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

  • Philippe BaumgartnerAffiliated withQuality, Health & Nutrition Group, Scottish Crop Research Institute
  • , Karen HarperAffiliated withQuality, Health & Nutrition Group, Scottish Crop Research Institute
  • , Romaan J.M. RaemaekersAffiliated withDepartment Agricultural and Environmental Sciences, King George VI Building, University of Newcastle
  • , Alain DurieuxAffiliated withUnité de Biotechnologie, Institut Meurice
  • , Angharad M.R. GatehouseAffiliated withDepartment Agricultural and Environmental Sciences, King George VI Building, University of Newcastle
  • , Howard V. DaviesAffiliated withQuality, Health & Nutrition Group, Scottish Crop Research Institute
  • , Mark A. TaylorAffiliated withQuality, Health & Nutrition Group, Scottish Crop Research Institute Email author 

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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