Food proteins from yeast-based precision fermentation: Simple purification of recombinant β-lactoglobulin using polyphosphate

L.J.G. Hoppenreijs, A. Annibal, G.J.C. Vreeke, R.M. Boom, J.K. Keppler*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Proteins produced through precision fermentation are often purified through chromatographic methods. Faster and more cost-effective purification methods are desired for food application. Here, we present a simple method for purification of protein produced from yeast, using β-lactoglobulin secreted from Pichia pastoris as an example. The food-grade salt hexametaphosphate (HMP) was used to precipitate the protein at acidic pH, while the impurities (extracellular polysaccharides; mainly mannan) remained soluble. After re-solubilization of the protein-HMP complex by neutralization, excess HMP was selectively precipitated using calcium chloride. The protein content of the crude sample increased from 26 to 72 wt% (comparable to purification with anion exchange chromatography), containing only residual extracellular polysaccharides (9 wt%) and HMP (1 wt%). The established method had no significant impact on the structural and functional properties (i.e., ability to form emulsions) of the protein. The presented method shows potential for cost-effective purification of recombinant proteins produced through yeast-based expression systems.

Original languageEnglish
Article number113801
JournalFood Research International
Volume176
DOIs
Publication statusPublished - Jan 2024

Keywords

  • Cellular agriculture
  • Extracellular polysaccharides
  • Graham's salt
  • Heterologous proteins
  • Komagataella phafii
  • Mannan
  • Pichia pastoris

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