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

Biotechnol Lett. 2003 Aug;25(15):1281-5. 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 1 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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bioreactors / microbiology*
  • Cell Culture Techniques / methods*
  • Galanthus / genetics
  • Galanthus / metabolism
  • Mannose-Binding Lectins / biosynthesis*
  • Mannose-Binding Lectins / chemistry*
  • Mannose-Binding Lectins / genetics
  • Molecular Sequence Data
  • Pichia / growth & development*
  • Pichia / metabolism*
  • Plant Lectins / biosynthesis*
  • Plant Lectins / chemistry*
  • Plant Lectins / genetics
  • Protein Engineering / methods*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification

Substances

  • Mannose-Binding Lectins
  • Plant Lectins
  • Recombinant Proteins
  • snowdrop lectin