The kinetic behavior of dehydrogenase enzymes in solution and immobilized onto nanostructured carbon platforms

Abstract

This paper describes the kinetic behavior of alcohol (ADH) and aldehyde (AldDH) dehydrogenases in solution and immobilized onto carbon platform via polyamidoamine (PAMAM) dendrimers. All the kinetic constants achieved for soluble ADH and AldDH are in agreement with literature data. The influence of pH and temperature was evaluated. Results showed that physiological conditions and ambient temperature can satisfactorily be applied to systems containing dehydrogenase enzymes, so as to ensure an environment where both ADH and AldDH display good activity. It is noteworthy that the affinity between both ADH and AldDH and their substrates and coenzyme is retained after the immobilization process. Investigation of the influence of the storage time demonstrated that there was no appreciable reduction in enzymatic activity for 50 days. Results showed that the PAMAM dendrimers provide a good environment for immobilization of dehydrogenase enzymes and that the affinity observed between the enzymes and their substrates and coenzymes seems to be retained, despite the considerable loss of enzymatic activity after immobilization. Furthermore, the anchoring methodology employed herein, namely layer-by-layer (LbL), required very low catalyst consumption.