Efficient brazzein production in yeast (Kluyveromyces lactis) using a chemically defined medium

Bioprocess Biosyst Eng. 2021 Apr;44(4):913-925. doi: 10.1007/s00449-020-02499-y. Epub 2021 Jan 27.

Abstract

The sweet-tasting protein brazzein offers considerable potential as a functional sweetener with antioxidant, anti-inflammatory, and anti-allergic properties. Here, we optimized a chemically defined medium to produce secretory recombinant brazzein in Kluyveromyces lactis, with applications in mass production. Compositions of defined media were investigated for two phases of fermentation: the first phase for cell growth, and the second for maximum brazzein secretory production. Secretory brazzein expressed in the optimized defined medium exhibited higher purity than in the complex medium; purification was by ultrafiltration using a molecular weight cutoff, yielding approximately 107 mg L-1. Moreover, the total media cost in this defined medium system was approximately 11% of that in the optimized complex medium to generate equal amounts of brazzein. Therefore, the K. lactis expression system is useful for mass-producing recombinant brazzein with high purity and yield at low production cost and indicates a promising potential for applications in the food industry.

Keywords: Brazzein; Chemically defined medium; Optimization of expression conditions; Sweet-tasting protein.

MeSH terms

  • Anti-Inflammatory Agents / chemistry
  • Antioxidants / chemistry
  • Biotechnology / methods
  • Culture Media
  • Densitometry
  • Fermentation
  • Hydrogen-Ion Concentration
  • Industrial Microbiology / methods
  • Kluyveromyces / metabolism*
  • Molecular Weight
  • Permeability
  • Plant Proteins / chemistry*
  • Recombinant Proteins / chemistry
  • Sweetening Agents / chemistry
  • Temperature

Substances

  • Anti-Inflammatory Agents
  • Antioxidants
  • Culture Media
  • Plant Proteins
  • Recombinant Proteins
  • Sweetening Agents
  • brazzein protein, Pentadiplandra brazzeana

Supplementary concepts

  • Kluyveromyces lactis