Presence of Anaplerotic Reactions and Transamination, and the Absence of the Tricarboxylic Acid Cycle in Mollicutes

1 Department of Medical Microbiology and Immunology, The Ohio State University, Columbus, Ohio 43210, USA 2 Division of Bacterial Products, National Center for Drugs and Biologics, Food and Drug Administration, Bethesda, Maryland 20205, USA ABSTRACT SUMMARY: Cell extracts of the fermentative Mollicutes Acholeplasma laidlawii B-PG9, Acholeplasma morum S2, Mycoplasma capricolum 14, Mycoplasma gallisepticum S6, Mycoplasma pneumoniae FH, Mycoplasma hyopneumoniae J and M. genitalium G-37, and the non-fermentative Mycoplasma hominis PG-21, Mycoplasma hominis 1620 and Mycoplasma bovigenitalium PG-11 were examined for 39 cytoplasmic enzyme activities associated with the tricarboxylic acid (TCA) cycle, transamination, anaplerotic reactions and other enzyme activities at the pyruvate locus. Malate dehydrogenase (EC 4.2.1.2 ) was the only TCA-cycle-associated enzyme activity detected and it was found only in the eight Mycoplasma species. Aspartate aminotransferase (EC 2.6.1.1 ) activity was detected in all Mollicutes tested except M. gallisepticum S6. Malate synthetase (EC 4.1.3.2 ) activity, in the direction of malate formation, was found in the eight Mycoplasma species, but not in any of the Acholeplasma species. Phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31 ) was detected in the direction of oxaloacetate (OAA) formation in both Acholeplasma species, but not in any of the Mycoplasma species. Pyruvate carboxylase (EC 6.4.1.1 ), pyruvate kinase (EC 2.7.1.40 ), pyruvate dehydrogenase (EC 1.2.4.1 ) and lactate dehydrogenase (EC 1.1.1.27 ) activities were found in all ten Mollicutes tested. No activities were detected in any of the ten Mollicutes for aspartase (EC 4.3.1.1 ), malic enzyme (EC 1.1.1.40 ), PEP carboxytransphosphorylase (EC 4.1.1.38 ), PEP carboxykinase (EC 4.1.1.32 ) or pyruvate orthophosphate dikinase (EC 2.7.9.1 ). In these TCA-cycle-deficient Mollicutes the pyruvate-OAA locus may be a point of linkage for the carbons of glycolysis, lipid synthesis, nucleic acid synthesis and certain amino acids. CO 2 fixation appears obligatory in the Acholeplasma species and either CO 2 fixation or malate synthesis appears obligatory in the Mycoplasma species.