Covalent Binding of an NAD Analogue to Liver Alcohol Dehydrogenase Resulting in an Enzyme‐Coenzyme Complex not Requiring Exogenous Coenzyme for Activity

摘要: 1 The NAD analogue, N6-[N-(6-aminohexyl)carbamoylmethyl]-NAD, was covalently bound to horse liver alcohol dehydrogenase in a carbodiimide-mediated reaction and such way that it active with the very same enzyme molecule which coupled. 2 The degree of substitution, i.e. number analogues per subunit, could be varied (0.3–1.6). In one preparation 1.6 coenzyme molecules were subunit; activity this 40% obtained after addition free excess. 3 It calculated every fourth site provided functioning analogue had cycling rate about 40000 cycles/h coupled substrate assay. 4 The presence made sites difficult inhibit competitive inhibitor. For example, 10 mM AMP inhibited by 50% whereas reference system containing native 80% spite fact contained 20000 times as high concentration coenzyme.