Metabolism of vitamin B-6 by human liver

The enzymes that metabolize vitamin B-6 were analyzed in liver biopsy samples from five patients without hepatic disease by using methods optimized for small samples. Pyridoxal kinase (EC 2.7.1.35) activities were 11.2 +/- 3.6 nmol/minute per gram of tissue and 0.16 +/- 0.05 nmol/minute per milligram of soluble protein (mean +/- SD); a clear dependence of the activity on zinc as the divalent cation was observed. Pyridoxine (pyridoxamine) 5'-phosphate oxidase (EC 1.4.3.5) activities, when using N-(5'-phosphopyridoxyl)-[3H]tryptamine as the substrate, were 0.64 +/- 0.22 pmol/minute per milligram of protein and 47 +/- 19 pmol/minute per gram of tissue. The activities were 63 +/- 18% lower when riboflavin 5'-phosphate was omitted from the assay; hence, it appears the oxidase is only partially saturated with its cofactor. The pyridoxal 5'-phosphate hydrolase(s) activities at alkaline pH were 282 +/- 183 nmol/minute per gram of tissue and 4.0 +/- 3.2 nmol/minute per milligram of particulate protein. Pyridoxal was rapidly oxidized to pyridoxic acid (28.1 +/- 19.8 nmol/minute per gram of tissue and 0.37 +/- 0.24 nmol/minute per milligram of soluble protein) by soluble enzyme(s), and the rate was unaffected by pyridine nucleotides. These experiments constitute the first quantitative analyses of the enzymes responsible for metabolizing vitamin B-6 in human liver, and provide data for interpreting the pharmacokinetics of B-6 utilization by humans, as well as methods for investigating diseases with aberrant metabolism of this nutrient.