Periplasmic metabolism of glutamate and aspartate by intact Bradyrhizobium japonicum bacteroids.
In studies on the uptake and metabolism of [14C]glutamate by Bradyrhizobium japonicum bacteroids we found that, in the presence of unlabeled malate, succinate or alpha-ketoglutarate, substantial label was recovered in alpha-ketoglutarate in the reaction mixtures. As much as 30% of the total 14C supplied could be found in alpha-ketoglutarate in the reaction mixtures after 30 min and this occurred in the absence of detectable labeling of alpha-ketoglutarate in the cells. The labeling of alpha-ketoglutarate was almost completely inhibited by aminooxyacetate (aminotransferase inhibitor). Direct assay of aspartate aminotransferase in intact bacteroids was possible in the presence of very dilute Triton X-100 (less than or equal to 0.02%, w/v). The response of the aminotransferase to detergent was similar to the response of phosphodiesterase, a periplasmic marker, and different from malate dehydrogenase and beta-hydroxybutyrate dehydrogenase, cytoplasmic markers. Comparison of maximum enzyme activity assayable with intact bacteroids and maximum activity in sonicated bacteroids indicated that about half of the total cellular aminotransferase activity was accessible to the external medium. The combined labeling and enzyme assay results indicated that B. japonicum bacteroids have a capability for transamination in the periplasmic space. Although this may not be important in the transfer of reducing equivalents from host cytoplasm to bacteroids in nodules, the transamination capability may facilitate the acquisition of metabolites by free-living bacteria.
Streeter
J G
JG
Department of Agronomy, Ohio State University, Wooster.
Salminen
S O
SO
D000693:Anaerobiosis; D001219:Aspartate Aminotransferases / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism*; D001692:Biological Transport; D002250:Carbon Radioisotopes; D003593:Cytoplasm / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D008293:Malates / Q000378:metabolism; D008954:Models, Biological; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D012230:Rhizobiaceae / Q000378:metabolism*
D002250:Carbon Radioisotopes; D005971:Glutamates; D008293:Malates; D001224:Aspartic Acid; D018698:Glutamic Acid; C030298:malic acid; D001219:Aspartate Aminotransferases; D010727:Phosphoric Diester Hydrolases