Background: UDP-galactopyranose mutase (UGM) catalyzes a step in galactofuranose biosynthesis in pathogens and is a promising drug design target. Results: The first crystal structures and SAXS analysis of UGM from the pathogenic fungus Aspergillus fumigatus are reported. Conclusion: The unique quaternary structure enables profound conformational changes to occur upon substrate binding. The structures support the covalent mechanism. Significance: The structures should aid inhibitor design.

Ion-exclusion high-pressure liquid chromatography (HPLC) was used to measure poly-f3-hydroxybutyrate (PHB) in Rhizobium japonicum bacteroids. The products in the acid digest of PHB-containing material were fractionated by HPLC on Aminex HPX-87H ion-exclusion resin for organic acid analysis. Crotonic acid formed from PHB during acid digestion was detected by its intense absorbance at 210 nm. The Aminex-HPLC method provides a rapid and simple chromatographic technique for routine analysis of organic acids. Results of PHB analysis by Aminex-HPLC were confirmed by gas chromatography and spectrophotometric analysis.

ABSTRACrThe activities of several enzymes of the citric acid and poly-4e-hydroxybutyrate cycles were measured in Rhizobiumjapeoicwm 3IIB-143 bacteroids which had been isolated from soybean nodules by sucrose gradient centrifupaton. Duing the period of developing nitrogenase activity, the specfic activity of fumuase, hydroxybutyrate dehydrogenase, ,-ketothiohlse, and pyruvate dehydrogenase complex incesed whereas acetoacetate-succinyl-CoA transferase and isocitrate dehydrogenase decreased (2,8,9,15,17,21,25,28,29).The carbon metabolism of developing and senescing nodules and the metabolism of bacteroids have been reviewed recently (5,19,27 PHB has been measured in many rhizobia species (7,12,14 The purpose of this investigation was, first, to establish that PDC, selected enzymes from the citric acid cycle, (IDH, MDH, and FMR) and of the PHB cycle (KT, HBD, and AST) were present during the period of nitrogen fixation and PHB accumulation in R. japonicum; second, to follow their activity as a function ofage; and third, to relate temporal changes in activities to those of nitrogenase and PHB accumulation through the use of dispersion diagrams.

Bradyrhizobium japonicum, the nitrogen-fixing symbiotic partner of soybean, was grown on various carbon substrates and assayed for the presence of the glyoxylate cycle enzymes, isocitrate lyase and malate synthase. The highest levels of isocitrate lyase [165-170 nmol min-1 (mg protein)-1] were found in cells grown on acetate or beta-hydroxybutyrate, intermediate activity was found after growth on pyruvate or galactose, and very little activity was found in cells grown on arabinose, malate, or glycerol. Malate synthase activity was present in arabinose- and malate-grown cultures and increased by only 50-80% when cells were grown on acetate. B. japonicum bacteroids, harvested at four different nodule ages, showed very little isocitrate lyase activity, implying that a complete glyoxylate cycle is not functional during symbiosis. The apparent Km of isocitrate lyase for D,L-isocitrate was fourfold higher than that of isocitrate dehydrogenase (61.5 and 15.5 microM, respectively) in desalted crude extracts from acetate-grown B. japonicum. When isocitrate lyase was induced, neither the Vmax nor the D,L-isocitrate Km of isocitrate dehydrogenase changed, implying that isocitrate dehydrogenase is not inhibited by covalent modification to facilitate operation of the glyoxylate cycle in B. japonicum.