Membrane entrapped Saccharomyces cerevisiae in a biosensor-like device as a generic rapid method to study cellular metabolism

J Biochem Biophys Methods. 2007 Apr 10;70(3):455-64. doi: 10.1016/j.jbbm.2006.11.001. Epub 2006 Nov 10.

Abstract

We describe a new, faster and convenient method to study some metabolic characteristics - by the successful application of immobilized yeast cells (S. cerevisiae) in a microbial biosensor-like device. Microbial biosensors consist of microorganisms immobilized on the surface of a membrane or in a gel, in close contact with a transducer. Almost all works published to date have used biosensors for analyses in which a concentration-related property of the external medium is measured. A different approach is presented here; we have successfully used S. cerevisiae and a carbon dioxide electrode as the main components of a biosensor-like device, used as a proof of concept, for a system useful to characterize metabolic parameters of the microbial cells immobilized on a carbon dioxide electrode. The biosensor-like device we are presenting allows us to calculate Michaelis-Menten parameters related to the kinetics of transport and degradation of several carbohydrates (i.e., glucose, fructose, galactose, sucrose and xylose, with K(m(app)) of 6.0, 5.8, 0.9, 2.0, and 147 mM, respectively), and the study of the kinetics of expression of non-constitutive proteins related to the transport and degradation of galactose.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / methods*
  • Carbohydrate Metabolism
  • Carbon Dioxide
  • Cells, Immobilized
  • Electrodes
  • Galactose / metabolism
  • Kinetics
  • Membrane Transport Proteins / metabolism
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / metabolism

Substances

  • Membrane Transport Proteins
  • Saccharomyces cerevisiae Proteins
  • Carbon Dioxide
  • Galactose