myo-lnositol catabolism and catabolite regulation in Rhizobium leguminosarum

Department of Microbiology, University of Reading, Whiteknights, PO Box 228, Reading RG6 ZAJ, UK On the basis of enzyme assays, myo-inositol appears to be catabolized via 2-keto-myo-inositol and ~-2,3-diketo-4-deoxy epi-inositol in Rhizobium leguminosarum bv. viciae, as occurs in Klebsiella aerogenes. The first two enzymes of the pathway, myo-inositol dehydrogenase and 2-keto-myo-inositol dehydratase were increased 7and 77-fold, respectively, after growth of R. leguminosarum on myo-inositol compared to glucose. Cells of R. leguminosarum grown on glucose as the carbon source and then resuspended in myo-inositol, increased myo-inositol-dependent 0, consumption by sixfold in 3 h, confirming this to be an inducible pathway. Succinate, malate and glucose exhibited strong catabolite repression of the myo-inositol catabolic pathway with myo-inositol dehydrogenase and 2-keto-myo-inositol dehydratase being repressed by a t least 68% and 8O%, respectively, in al l cases. A dicarboxylate transport mutant (dctA) was unable to repress either enzyme when grown on myo-inositol and succinate. There was no repression of the first two enzymes of the myo-inositol catabolic pathway in the background of constitutive expression of the dicarboxylate transport system, indicating a dicarboxylate must be present to cause repression. The data imply that dicarboxylates are required intra-cellularly to repress these enzymes. Three transposon mutants were isolated that cannot grow on myo-inositol as the sole carbon source and were unable to induce either myo-inositol dehydrogenase or 2-keto-myo-inositol dehydratase. The mutants were unaffected in the ability to nodulate peas and vetch. Furthermore, vetch plants infected with one mutant (RU360) reduced acetylene at the same rate as plants infected with the wild type. Bacteroids did not oxidize myo-inositol, nor were the catabolic enzymes detected, confirming myo-inositol is not important as an energy source in bacteroids. The possible role of myo-inositol in the rhizosphere is considered.