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An apparent direct electron transfer between various electrode materials and peroxidases immobilized on the surface of the electrode has been reported in the last few years. An electrocatalytic reduction of hydrogen peroxide stars at about +600 mV versus a saturated calomel (reference) electrode (SCE) at neutral pH. The efficiency of the electrocatalytic current increases as the applied potential is made more negative and starts t o level off at about -200 mV versus SCE. Amperometric biosensors for hydrogen peroxide can be constructed with these types of peroxidase modified electrodes. By co-immobilizing a hydrogen peroxide-producing oxidase with the peroxidase, amperometric biosensors can be made that respond t o the substrate of the oxidase within a potential range essentially free of interfering electrochemical reactions. Examples of glucose, alcohol and amino acid sensors are shown.

The main contribution of this paper is a new "extrinsic" digital fundamental group that can be readily generalized to define higher homotopy groups for arbitrary digital spaces. We show that the digital fundamental group of a digital object is naturally isomorphic to the fundamental group of its continuous analogue. In addition, we state a digital version of the Seifert-Van Kampen theorem.

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Abstract. In this paper a notion of lighting function is introduced as an axiomatized formalization of the "face membership ruleg' suggested by Kovalevsky. These functions are defined in the context of the framework for digital topology previously developed by the authors. This enlarged framework provides the (a, fl)-connectedness (~, fl E {6, 18, 26}) defined on 2[ 3 within the graph-based approach to digital topology. Furthermore, the Kong-Roscoe (c~,fl)-surfaces, with (a,~) 5~ (6,6), (18,6), are also found as particular cases of a more general notion of digital surface.

This work is aimed towards the generation of enzyme arrays on electrochemically active surfaces by taking advantage of the DNA-directed immobilization (DDI) technique. To this end, two different types of horseradish peroxidase (HRP)-DNA conjugates were prepared, either by covalent coupling with a bifunctional cross-linker or by the reconstitution of apo-HRP, that is, HRP lacking its prosthetic heme (protoporphyrin IX) group, with a covalently DNA-modified heme cofactor. Both conjugates were characterized in bulk and also subsequent to their immobilization on gold electrodes through specific DNA hybridization. Electrochemical measurements by using the phenolic mediator ortho-phenylendiamine indicated that, due to the high degree of conformational orientation, the apparent Michaelis-Menten constants of the reconstituted HRP conjugate were lower than those of the covalent conjugate. Due to the reversible nature of DDI, both conjugates could be readily removed from the electrode surface by simple washing and, subsequently, the electrodes could be reloaded with fresh enzymes, thereby restoring the initial amperometric-response activity. Moreover, the specific DNA hybridization allowed us to direct the two conjugates to distinct sites on a microelectrode array. Therefore, the self-assembly and regeneration capabilities of this approach should open the door to the generation of arrays of redox-enzyme devices for the screening of enzymes and their effectors.