Concanavalin A-agarose removes mannan impurities from an extracellularly expressed Pichia pastoris recombinant protein

Cell Mol Biol Lett. 2003;8(3):783-92.

Abstract

Pichia pastoris secretes few native proteins. However, the more than 1 g l(-1)of extracellularly expressed mannan interfered with the purification of our extracellularly expressed, non-glycosylated recombinant protein. Concanavalin A-agarose removed more than 95% of the unwanted mannan as monitored by phenol reaction. A(13)C-based NMR assay confirmed this improvement. Concanavalin A-agarose can assist the purification of extracellular expressed, non-glycosylated proteins from yeasts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Isotopes
  • Chromatography, Ion Exchange
  • Concanavalin A / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression Regulation, Fungal
  • Magnetic Resonance Spectroscopy
  • Mannans / isolation & purification*
  • Pichia / metabolism*
  • Plant Proteins / analysis
  • Plant Proteins / metabolism
  • Polysaccharides / analysis
  • Recombinant Proteins / metabolism*
  • Sepharose / metabolism*

Substances

  • Carbon Isotopes
  • Mannans
  • Plant Proteins
  • Polysaccharides
  • Recombinant Proteins
  • Concanavalin A
  • Sepharose