Encapsulation of Microorganisms, Enzymes, and Redox Mediators in Graphene Oxide and Reduced Graphene Oxide

Graphene oxide (GO) and reduced graphene oxide (rGO) were demonstrated in the past decade as biocompatible carbon-based materials that could be efficiently used in bioelectrochemical systems (BESs). Specifically, for redox enzyme encapsulation in order to improve electron communication between enzymes and electrodes. The addition of GO to different solvents was shown to cause gelation while still allowing small molecule diffusion through its gel-like matrix. Taking the combination of these traits together, we decided to use GO hydrogels for the encapsulation of enzymes displayed on the surface of yeast in anodes of microbial fuel cells. During our studies we have followed the changes in the physical characteristics of GO upon encapsulation of yeast cells displaying glucose oxidase in the presence of glucose and noted that GO is being rapidly reduced to rGO as a function of glucose concentrations. GO reduction under these conditions served as a proof of electron communication between the surface-displayed enzymes and GO. Hence, we set out to study this phenomenon by the encapsulation of a purified glucose dehydrogenase (in the absence of microbial cells) in rGO where improved electron transfer to the electrode could be observed in the presence of phenothiazone. In this chapter, we describe how these systems were technically constructed and characterized and how a very affordable matrix such as GO could be used to electrically wire enzymes as a good replacement for expensive mediator containing redox active polymers commonly used in BESs.