Membrane entrapped Saccharomyces cerevisiae in a biosensor-like device as a generic rapid method to study cellular metabolism.
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.
Martínez
Mariela
M
Department of Biochemistry, School of Science, University of Buenos Aires, Ciudad Universitaria, Pab. 2, (1428), Buenos Aires, Argentina.
Hilding-Ohlsson
Astrid
A
Viale
Alberto A
AA
Cortón
Eduardo
E
D015374:Biosensing Techniques / Q000379:methods*; D050260:Carbohydrate Metabolism; D002245:Carbon Dioxide; D018914:Cells, Immobilized; D004566:Electrodes; D005690:Galactose / Q000378:metabolism; D007700:Kinetics; D026901:Membrane Transport Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
D026901:Membrane Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; D002245:Carbon Dioxide; D005690:Galactose