The sensing of enzymatic processes in volumes at or below the scale of single cells is challenging but highly desirable in the study of biochemical processes. Here we demonstrate a nanofluidic device which combines an enzymatic recognition element and electrochemical signal transduction within a six femtoliter volume. Our approach is based on localized immobilization of the enzyme tyrosinase in a microfabricated nanogap electrochemical transducer. The enzymatic reaction product quinone is localized in the confined space of a nanochannel in which efficient redox cycling also takes place. Thus the sensor allows the sensitive detection of minute amounts of product molecules generated by the enzyme in real time. This method is ideally suited for the study of ultra-small volume systems such as the contents of individual biological cells or organelles.