DNA sequencing and expression of the formyl coenzyme A transferase gene, frc, from Oxalobacter formigenes.
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 CO2. 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 rho-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.
Sidhu
H
H
Program in Experimental Pathology, Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville 32610, USA.
Ogden
S D
SD
Lung
H Y
HY
Luttge
B G
BG
Baetz
A L
AL
Peck
A B
AB
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D015256:Coenzyme A-Transferases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D004789:Enzyme Activation / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data
D001426:Bacterial Proteins; D015256:Coenzyme A-Transferases; C064813:formyl-coenzyme A transferase