An optical biosensor for lysine based on the use of lysine decarboxylase and a cadaverine-sensitive membrane

Abstract

We describe an optical biosensor for lysine based on the use of lysine decarboxylase and an optical transducer for detection of cadaverine which is formed as a result of enzymatic action. A plasticized PVC (polyvinyl chloride) membrane containing a lipophilic tartrate as the amine carrier acts as the optical cadaverine sensor. The transport of the cadaverine cation into the membrane is coupled to a transport of a proton (of the indicator dye) out of the membrane. This causes a spectral change of the indicator dye which can be related to the cadaverine concentration, provided the pH is kept constant. The enzymatic reaction is performed in an enzyme reactor which is part of a flow-through system. The dynamic range is from 0·1 to 100 mM for both cadaverine and lysine. While the cadaverine sensor is moderately selective (ethylamines, for example, interfere), the whole sensor system is highly specific for lysine, nicotine being the only major interferent. Unlike other enzyme-based detection schemes where the production of CO2 (in case of decarboxylates) or consumption of oxygen (in case of oxidases) is measured, this scheme is based on the measurement of the organic ammonium ion (cadaverin cation) formed in the enzymatic reaction. The major advantage of this approach is that in many real samples there is a rather low and fairly constant background of organic amines. This is in contrast to sensors based on the measurements of oxygen consumption (in the case of amino acid oxidases) or carbon dioxide production (using decarboxylases), where the background levels of the respective gases (which are ubiquitous) have to be kept constant in order to specifically measure only the concentration changes caused by the enzymatic reaction, or need to be measured in an independent assay. © 1992.