Critical evaluation of sampling techniques for residual glucose determination in carbon-limited chemostat culture of Saccharomyces cerevisiae

Biotechnol Bioeng. 2003 Aug 20;83(4):395-9. doi: 10.1002/bit.10683.

Abstract

In this paper, three sampling techniques for rapid quenching of cellular metabolism and subsequent separation of cells from fermentation broth are compared: (i) quick freezing of fermentation broth directly in liquid nitrogen; (ii) quenching metabolism by exposing the fermentation broth to stainless steel beads (4-mm diameter) in a filter syringe precooled to -18 degrees C; and (iii) withdrawal of the filtrate through a 0.45 microm filter attached to a syringe and a needle inserted directly into the fermentor. It was concluded that use of liquid nitrogen as a quenching method to rapidly arrest cellular metabolism, for quantitative analysis of extracellular glucose, is not a very reliable method and that the filter syringe steel beads work very well.

Publication types

  • Comparative Study
  • Evaluation Study
  • Validation Study

MeSH terms

  • Bioreactors / microbiology*
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Cold Temperature
  • Culture Media / analysis*
  • Culture Media / radiation effects
  • Freezing
  • Glucose / analysis*
  • Glucose / metabolism*
  • Reproducibility of Results
  • Saccharomyces / chemistry
  • Saccharomyces / growth & development*
  • Saccharomyces / metabolism*
  • Saccharomyces / radiation effects
  • Sensitivity and Specificity

Substances

  • Culture Media
  • Glucose