The molecular mechanism of flavoprotein catalysis, with special emphasis on the role of substrate carbanions and their covalent adducts with the flavin nucleus, will be further investigated using model carbanionic substrates and derivatives of physiological substrates. Derivatized adenine nucleotide coenzymes, containing primary amino and carboxyl functions will be coupled chemically to soluble and insoluble carriers. The immobilized coenzymes which result wll be tested for enzymatic reactivity in single- and multi-enzyme systems.