Development of a multi‐parameter sensor chip for the simultaneous detection of organic compounds in biogas processes

An enzyme-based multi-parameter biosensor is developed for monitoring the concentration of formate, d-lactate, and l-lactate in biological samples. The sensor is based on the specific dehydrogenation by an oxidized β-nicotinamide adenine dinucleotide (NAD+)-dependent dehydrogenase (formate dehydrogenase, d-lactic dehydrogenase, and l-lactic dehydrogenase, respectively) in combination with a diaphorase from Clostridium kluyveri (EC 1.8.1.4). The enzymes are immobilized on a platinum working electrode by cross-linking with glutaraldehyde (GA). The principle of the determination scheme in case of l-lactate is as follows: l-lactic dehydrogenase (l-LDH) converts l-lactate into pyruvate by reaction with NAD+. In the presence of hexacyanoferrate(III), the resulting reduced β-nicotinamide adenine dinucleotide (NADH) is then regenerated enzymatically by diaphorase. The electrochemical detection is based on the current generated by oxidation of hexacyanoferrate(II) at an applied potential of +0.3 V vs. an Ag/AgCl reference electrode. The biosensor will be electrochemically characterized in terms of linear working range and sensitivity. Additionally, the successful practical application of the sensor is demonstrated in an extract from maize silage.

[1]  Michael J. Schöning,et al.  New concepts of miniaturised reference electrodes in silicon technology for potentiometric sensor systems , 2004 .

[2]  M. Comtat,et al.  Amperometric bienzyme electrode for l-carnitine , 1988 .

[3]  Sang Hyun Lee,et al.  Immobilization of formate dehydrogenase from Candida boidinii through cross-linked enzyme aggregates , 2013 .

[4]  T. Noguer,et al.  An amperometric bienzyme electrode for acetaldehyde detection , 1995 .

[5]  Rasa Pauliukaite,et al.  An improved biosensor for acetaldehyde determination using a bienzymatic strategy at poly(neutral red) modified carbon film electrodes. , 2007, Analytica chimica acta.

[6]  P. Weiland.  Biogas production: current state and perspectives , 2009, Applied Microbiology and Biotechnology.

[7]  Jean-Louis Marty,et al.  Chronoamperometric determination of d-lactate using screen-printed enzyme electrodes , 2001 .

[8]  Richard S. Nicholson,et al.  Theory and Application of Cyclic Voltammetry for Measurement of Electrode Reaction Kinetics. , 1965 .

[9]  A. Legin,et al.  Development of a thin‐film sensor array for analytical monitoring of heavy metals in aqueous solutions , 2012 .

[10]  Isabelle Migneault,et al.  Glutaraldehyde: behavior in aqueous solution, reaction with proteins, and application to enzyme crosslinking. , 2004, BioTechniques.

[11]  Ioanis Katakis,et al.  Catalytic electrooxidation of NADH for dehydrogenase amperometric biosensors , 1997 .

[12]  C Hung Tzang,et al.  Voltammetric biosensors for the determination of formate and glucose-6-phosphate based on the measurement of dehydrogenase-generated NADH and NADPH. , 2001, Biosensors & bioelectronics.

[13]  J. Schubert,et al.  Development of redox glasses and subsequent processing by means of pulsed laser deposition for realizing silicon-based thin-film sensors , 2013 .

[14]  G. Broun.  [20] Chemically aggregated enzymes , 1976 .

[15]  B. Ahring,et al.  Regulation and optimization of the biogas process: Propionate as a key parameter , 2007 .

[16]  R. Meeske,et al.  The effect of a lactic acid bacterial inoculant on maize silage , 1998 .

[17]  Ulla Wollenberger,et al.  An amperometric bi-enzyme sensor for determination of formate using cofactor regeneration. , 2003, Biosensors & bioelectronics.

[18]  Robert F. Hickey,et al.  Monitoring of the anaerobic methane fermentation process , 1990 .

[19]  M. Comtat,et al.  Bienzyme amperometric lactate-specific electrode , 1990 .

[20]  B. Bachmann,et al.  Micromachined multi‐parameter sensor chip for the control of polymer‐degradation medium , 2014 .

[21]  Dario Compagnone,et al.  Recent advances in NADH electrochemical sensing design. , 2009, Bioelectrochemistry.

[22]  Miltiades I. Karayannis,et al.  Enzyme Based Amperometric Biosensors for Food Analysis , 2002 .

[23]  P. Tuñón,et al.  Amperometric biosensors based on NAD(P)‐dependent dehydrogenase enzymes , 1997 .

[24]  Simona Serban,et al.  Redox-flexible NADH oxidase biosensor: A platform for various dehydrogenase bioassays and biosensors , 2006 .

[25]  Audrey Sassolas,et al.  Immobilization strategies to develop enzymatic biosensors. , 2012, Biotechnology advances.

[26]  Jean-Louis Marty,et al.  Comparison of the performances of two bi-enzymatic sensors for the detection of D-lactate , 1995 .