Oxidation of glycine by Pseudomonas putida requires a specific lipoamide dehydrogenase

Abstract

Pseudomonas putida produces two lipoamide dehydrogenases with molecular weights of 49,000 and 56,000 designated LPD-val and LPD-glc, respectively. LPD-val is required for oxidation of valine, since it is specifically utilized as the E3 component of branched-chain keto acid dehydrogenase. Since glycine oxidation by bacteria and mammals also requires lipoamide dehydrogenase, we desired to determine which lipoamide dehydrogenase would be used by the P. putida glycine oxidation system. When grown in a medium with glycine as the sole nitrogen source, P. putida produced a single lipoamide dehydrogenase with a molecular weight of 56,000 and which reacted with antiserum to LPD-glc. The partially purified glycine oxidation system from P. putida was stimulated by LPD-glc but not by LPD-val and was inhibited by anti-LPD-glc, but not by anti-LPD-val. It was not possible to detect LPD-val in extracts of cells grown in glucose-glycine medium by the use of anti-LPD-val. LPD-glc was five times as active as LPD-val in catalyzing the oxidation of purified protein H, the heat-stable, lipoic acid-containing protein of the glycine oxidation system. These results indicate that LPD-glc is specifically utilized for glycine oxidation in P. putida.