Improving enzyme-electrode contacts by redox modification of cofactors.
Efficient electron transfer of redox proteins to and from their environment is essential for the use of such proteins in biotechnological applications such as amperometric biosensors and photosynthetic biocatalysts. But most redox enzymes lack pathways that can transport an electron from their embedded redox site to an electrode or a diffusing photoexcited species. Electrical communication between redox proteins and electrode surfaces has been improved by aligning proteins on chemically modified electrodes, by attaching electron-transporting groups and by immobilizing proteins in polymer matrices tethered by redox groups. Generally these methods involve contacting the enzymes at random with electron relay units. Here we report an approach that allows site-specific positioning of electron-mediating units in redox proteins. We strip glucose oxidase of its flavin adenine dinucleotide (FAD) cofactors, modify the latter with redox-active ferrocene-containing groups, and then reconstitute the apoprotein with these modified cofactors. In this way, electrical contact between an electrode and the resulting enzyme in solution is greatly enhanced in a controlled and reproducible way.
Riklin
A
A
Institute of Chemistry, Hebrew University of Jerusalem, Israel.
Katz
E
E
Willner
I
I
Stocker
A
A
Bückmann
A F
AF
D001059:Apoproteins / Q000737:chemistry*; D002384:Catalysis; D003605:D-Amino-Acid Oxidase / Q000737:chemistry; D004563:Electrochemistry; D004566:Electrodes; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D005296:Ferrous Compounds / Q000737:chemistry; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry*; D005949:Glucose Oxidase / Q000737:chemistry*; D010084:Oxidation-Reduction
D001059:Apoproteins; D005296:Ferrous Compounds; D005182:Flavin-Adenine Dinucleotide; D005949:Glucose Oxidase; D003605:D-Amino-Acid Oxidase; D005022:Ethyldimethylaminopropyl Carbodiimide