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Title: DNA SEQUENCING AND EXPRESSION OF THE FORMYL COENZYME A TRANSFERASE GENE, FRC, FROM OXALOBACTER FORMIGENES

Author
item SIDHU, H - UNIVERSITY OF FLORIDA
item OGDEN, S - IXION BIOTECHNOLOGY, FL
item LUNG, H - UNIVERSITY OF FLORIDA
item LUTTGE, B - UNIVERSITY OF FLORIDA
item Baetz, Albert
item PECK, A - UNIVERSITY OF FLORIDA

Submitted to: Journal of Bacteriology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/6/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary: Kidney stones caused by calcium oxalate are a continuing health problem to farm animals and humans. The enzymes we found in Oxalobacter formigenes degrade oxalate and make it nontoxic. In this study, the sequence of the gene for formyl-CoA transferase was determined and its location on the bacterial chromosome was found. This should lead to the cloning of this gene into other bacteria found in the digestive tract of man and farm animals, and ultimately to the alleviation of the problem of kidney stones by oxalate. This would alleviate the pain, suffering, and the expense associated with treating and removing kidney stones. This information would be used by both the medical and veterinary profession.

Technical Abstract: Oxalic acid, a highly toxic by-product of metabolism, is catabolized by a limited number of bacterial species utilizing an activation-decarboxylation reacton which yields formate and CO2. Frc, the gene encoding formyl-CoA transferase, an enzyme which transfers a Coenzyme A moiety to activate oxalic acid, was cloned from the bacterium Oxlobacter formigenes. The DNA sequence revealed a single open reading frame of 1284 base pairs capable o encoding a 428 amino acid protein. A presumed promoter region and a rho-in dent termination sequence suggests 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.