Genetic modification of the marine-derived yeast Yarrowia lipolytica with high-protein content using a GPI-anchor-fusion expression system

Biotechnol Prog. 2009 Sep-Oct;25(5):1297-303. doi: 10.1002/btpr.235.

Abstract

Yarrowia lipolytica SWJ-1b isolated from the marine fish gut was found to contain 47.6 g of crude protein per 100 g of cell dry weight and had potential use as single cell protein. When the gene encoding enhanced green fluorescent protein (EGFP) was inserted into the surface display plasmid pINA1317-YlCWP110 and expressed in uracil mutant of Y. lipolytica SWJ-1b, the corresponding protein was successfully displayed on the cell surface, and 100% of the yeast cells exhibited the anchored target proteins. We found that yeast cells displaying EGFP were similar to those of Y. lipolytica SWJ-1b. Furthermore, C(18:1) and C(18:3) fatty acids biosynthesis in the marine yeast cells displaying the heterologous EGFP was weakly enhanced compared with that in its wild-type. The results suggest that the marine-derived Y. lipolytica SWJ-1b can be armed with the heterologous protein by the genetic modification and further used as single cell protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / analysis
  • Cloning, Molecular / methods*
  • Fatty Acids / analysis
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Glycosylphosphatidylinositols / genetics*
  • Glycosylphosphatidylinositols / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Polymerase Chain Reaction
  • Protein Engineering / methods*
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / metabolism
  • Uracil / metabolism
  • Yarrowia / cytology
  • Yarrowia / genetics*
  • Yarrowia / metabolism

Substances

  • Amino Acids
  • Fatty Acids
  • Fungal Proteins
  • Glycosylphosphatidylinositols
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Uracil