VITAMIN-B12 DEFICIENCY INCREASES THE SPECIFIC ACTIVITIES OF RAT-LIVER NADH-LINKED AND NADPH-LINKED AQUACOBALAMIN REDUCTASE ISOZYMES INVOLVED IN COENZYME SYNTHESIS

Rat liver contains both NADH- and NADPH-linked aquacobalamin reductases, which are involved in the synthesis of the vitamin B-12 coenzymes and are distributed in both the mitochondrial and microsomal membranes. To clarify the physiological roles of these hepatic enzymes, vitamin B-12-deficient rats were used to study the effect of the deficiency on the enzyme activities. Male rats fed a vitamin B-12-deficient diet for 11 wk developed a severe vitamin B-12 deficiency with a high urinary methylmalonate excretion (214.3 +/- 115.2-mu-mol/d) and approximately 96% lower hepatic vitamin B-12 content. Tissues of the vitamin B-12-deficient rats were assayed for NADH- and NADPH-linked aquacobalamin reductase activities. The specific activities of both enzymes in homogenates of liver, kidney or upper intestine were shown to be three- to 20-fold greater in the vitamin-deficient rats than in the control rats. In liver, the vitamin deficiency specifically elevated the specific activities of the mitochondrial NADH-linked and microsomal NADPH-linked enzymes. These are likely the isozymes involved in vitamin B-12 coenzyme synthesis.