Detection of two distinct substrate-dependent catabolic responses in yeast cells using a mediated electrochemical method

Abstract. Mediated electrochemical detection of catabolism in prokaryotic cells is well documented; however, the application of this technique to eukaryotic cells has received less attention. Two catabolic substrate-dependent mediated electrochemical signals were detected in the yeast Saccharomyces cerevisiae. The signal using a single hydrophilic mediator (ferricyanide) is small whereas the response using a double mediator system comprising a hydrophilic and a lipophilic mediator (ferricyanide and menadione) is up to three orders of magnitude larger. The behaviour of each response during cell ageing is different: the single mediator response increases whereas the double mediator response decreases. This difference indicates that the two signals originate at different points in the catabolic pathways. In S. cerevisiae the double mediator response is proposed to originate from the reduction of the lipophilic mediator by NADPH produced in the pentose phosphate pathway. The single mediator signal arises from reduction of the hydrophilic mediator by an extracellular redox species produced in response to the presence of glucose.

[1]  Anthony Turner,et al.  Development of an electrochemical method for the rapid determination of microbial concentration and evidence for the reaction mechanism , 1988 .

[2]  R. Renneberg,et al.  Measurement of biodegradable substances using the salt-tolerant yeast Arxula adeninivorans for a microbial sensor immobilized with poly(carbamoyl) sulfonate (PCS) Part I: Construction and characterization of the microbial sensor. , 1999, Biosensors & bioelectronics.

[3]  Graeme M. Walker,et al.  Yeast Physiology and Biotechnology , 1998 .

[4]  D. Rawson,et al.  The development of whole cell biosensors for on‐line screening of herbicide pollution of surface waters , 1987 .

[5]  N. Pasco,et al.  Biochemical mediator demand – a novel rapid alternative for measuring biochemical oxygen demand , 2000, Applied Microbiology and Biotechnology.

[6]  Richard A. Durst,et al.  Mediator compounds for the electrochemical study of biological redox systems: a compilation , 1982 .

[7]  R. Renneberg,et al.  Measurement of biodegradable substances using the salt-tolerant yeast Arxula adeninivorans for a microbial sensor immobilized with poly(carbamoyl)sulfonate (PCS). Part II: Application of the novel biosensor to real samples from coastal and island regions. , 1999, Biosensors & bioelectronics.

[8]  B. D. Malhotra,et al.  Mediated biosensors. , 2002, Biosensors & bioelectronics.

[9]  Wei Cheng,et al.  Scanning electrochemical microscopy of living cells Part 2. Imaging redox and acid:basic reactivities , 2001 .

[10]  J. Luong,et al.  Mediated microbial biosensor using a novel yeast strain for wastewater BOD measurement , 2001, Applied Microbiology and Biotechnology.

[11]  P Ertl,et al.  Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes. , 2000, Analytical chemistry.

[12]  K. Bayer,et al.  Ferricyanide reduction by Escherichia coli: kinetics, mechanism, and application to the optimization of recombinant fermentations. , 2000, Analytical chemistry.

[13]  H. Mcconnell,et al.  Potentiometric Measurement of Intracellular Redox Activity , 1998 .

[14]  James F Rusling,et al.  Scanning electrochemical microscopy of living cells. 3. Rhodobacter sphaeroides. , 2002, Analytical chemistry.

[15]  Koon Gee Neoh,et al.  Microbial membrane-modified dissolved oxygen probe for rapid biochemical oxygen demand measurement , 1992 .

[16]  David M. Rawson,et al.  A chemically mediated amperometric biosensor for monitoring eubacterial respiration , 1991 .

[17]  P Ertl,et al.  Electrochemical biosensor array for the identification of microorganisms based on lectin-lipopolysaccharide recognition. , 2001, Analytical chemistry.

[18]  Hirohisa Tanaka,et al.  BOD BIOSENSOR FOR SECONDARY EFFLUENT FROM WASTEWATER TREATMENT PLANTS , 1994 .

[19]  Amperometric Biochemical Characterization of Isolated Fungal Strains , 2001 .