Application of cell-surface engineering for visualization of yeast in bread dough: development of a fluorescent bio-imaging technique in the mixing process of dough

Biosci Biotechnol Biochem. 2009 Jul;73(7):1604-7. doi: 10.1271/bbb.90106. Epub 2009 Jul 7.

Abstract

Cell-surface engineering (Ueda et al., 2000) has been applied to develop a novel technique to visualize yeast in bread dough. Enhanced green fluorescent protein (EGFP) was bonded to the surface of yeast cells, and 0.5% EGFP yeasts were mixed into the dough samples at four different mixing stages. The samples were placed on a cryostat at -30 degrees C and sliced at 10 microm. The sliced samples were observed at an excitation wavelength of 480 nm and a fluorescent wavelength of 520 nm. The results indicated that the combination of the EGFP-displayed yeasts, rapid freezing, and cryo-sectioning made it possible to visualize 2-D distribution of yeast in bread dough to the extent that the EGFP yeasts could be clearly distinguished from the auto-fluorescent background of bread dough.

MeSH terms

  • Biotechnology / methods
  • Bread / microbiology*
  • Fluorescent Dyes / metabolism*
  • Food Handling*
  • Freezing
  • Green Fluorescent Proteins / metabolism*
  • Luminescent Measurements / methods*
  • Microscopy, Fluorescence
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / isolation & purification*
  • Saccharomyces cerevisiae / metabolism
  • Time Factors

Substances

  • Fluorescent Dyes
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins