DNA sequencing and expression of the formyl coenzyme A transferase gene, frc, from Oxalobacter formigenes

Abstract

Oxalic acid, a highly toxic by-product of metabolism, is catabolized by a limited number of bacterial species utilizing an activation-decarboxylation reaction which yields formate and CO 2 . frc, the gene encoding formyl coenzyme A transferase, an enzyme which transfers a coenzyme A moiety to activate oxalic acid, was cloned from the bacterium Oxalobacter formigenes. DNA sequencing revealed a single open reading frame of 1,284 bp capable of encoding a 428-amino-acid protein. A presumed promoter region and a -independent termination sequence suggest that this gene is part of a monocistronic operon. A PCR fragment containing the open reading frame, when overexpressed in Escherichia coli, produced a product exhibiting enzymatic activity similar to the purified native enzyme. With this, the two genes necessary for bacterial catabolism of oxalate, frc and oxc, have now been cloned, sequenced, and expressed.

Insights into the lifestyle of uncultured bacterial natural product factories associated with marine sponges

“…Entotheonella could use oxalate as an energy source. Additionally, a putative oxalate/formate antiporter (ETW98700 and ETX08188) could directly generate a proton motive force similar as described for Oxalobacter formigenes (34). Notably, the formation of calcium oxalate has been reported in the marine sponge Chondrosia reniformis (35).…”

A Link between Arabinose Utilization and Oxalotrophy in Bradyrhizobium japonicum

“…1) were expressed abundantly in minimal medium containing L-arabinose. Remarkably, large amounts of two enzymes involved in oxalate degradation were also detected: a formyl-CoA transferase (Frc) (bll3156) and an oxalylCoA decarboxylase (Oxc) (bll3157) that share 71% and 78% amino acid sequence identity with the previously studied Frc and Oxc of Oxalobacter formigenes (39)(40)(41). This led us to speculate that glycolaldehyde is oxidized to glyoxylate, which can be fed into at least two pathways: (i) reduction to glycerate through the activity of glyoxylate carboligase (blr3166) (42) and tartronate semialdehyde reductase (blr3168), or (ii) oxidation to oxalate followed by the stepwise, complete oxidation to formate and CO 2 through the activities of Oxc and Frc (43,44) and formate dehydrogenase.…”

Succinyl-CoA:( R )-Benzylsuccinate CoA-Transferase: an Enzyme of the Anaerobic Toluene Catabolic Pathway in Denitrifying Bacteria

“…The catalytic mechanisms of these enzymes require ternary-complex formation of the enzyme, donor ACP-thioester (acetyl-ACP), and a thioester-acceptor compound (citrate or citramalate; 10, 11). (iii) Finally, the first members of a third family of CoA-transferases were recently characterized: (E)-cinnamoyl-CoA:(R)-phenyllactate CoAtransferase from Clostridium sporogenes (16) and formyl-CoA: oxalate CoA-transferase from Oxalobacter formigenes (3,32). These enzymes differ in sequence from the enzymes of families I and II, and a reaction mechanism similar to that of family II enzymes has been suggested for one member (16).…”