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. 1997 May;179(10):3378–3381. doi: 10.1128/jb.179.10.3378-3381.1997

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

H Sidhu 1, S D Ogden 1, H Y Lung 1, B G Luttge 1, A L Baetz 1, A B Peck 1
PMCID: PMC179125  PMID: 9150242

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 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.

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Selected References

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  1. Abe K., Ruan Z. S., Maloney P. C. Cloning, sequencing, and expression in escherichia coli of OxlT, the oxalate:formate exchange protein of Oxalobacter formigenes. J Biol Chem. 1996 Mar 22;271(12):6789–6793. doi: 10.1074/jbc.271.12.6789. [DOI] [PubMed] [Google Scholar]
  2. Allison M. J., Cook H. M., Milne D. B., Gallagher S., Clayman R. V. Oxalate degradation by gastrointestinal bacteria from humans. J Nutr. 1986 Mar;116(3):455–460. doi: 10.1093/jn/116.3.455. [DOI] [PubMed] [Google Scholar]
  3. Allison M. J., Dawson K. A., Mayberry W. R., Foss J. G. Oxalobacter formigenes gen. nov., sp. nov.: oxalate-degrading anaerobes that inhabit the gastrointestinal tract. Arch Microbiol. 1985 Feb;141(1):1–7. doi: 10.1007/BF00446731. [DOI] [PubMed] [Google Scholar]
  4. Anantharam V., Allison M. J., Maloney P. C. Oxalate:formate exchange. The basis for energy coupling in Oxalobacter. J Biol Chem. 1989 May 5;264(13):7244–7250. [PubMed] [Google Scholar]
  5. Baetz A. L., Allison M. J. Purification and characterization of formyl-coenzyme A transferase from Oxalobacter formigenes. J Bacteriol. 1990 Jul;172(7):3537–3540. doi: 10.1128/jb.172.7.3537-3540.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Baetz A. L., Allison M. J. Purification and characterization of oxalyl-coenzyme A decarboxylase from Oxalobacter formigenes. J Bacteriol. 1989 May;171(5):2605–2608. doi: 10.1128/jb.171.5.2605-2608.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clark J. H., Fitzgerald J. F., Bergstein J. M. Nephrolithiasis in childhood inflammatory bowel disease. J Pediatr Gastroenterol Nutr. 1985 Oct;4(5):829–834. doi: 10.1097/00005176-198510000-00026. [DOI] [PubMed] [Google Scholar]
  8. Dawson K. A., Allison M. J., Hartman P. A. Isolation and some characteristics of anaerobic oxalate-degrading bacteria from the rumen. Appl Environ Microbiol. 1980 Oct;40(4):833–839. doi: 10.1128/aem.40.4.833-839.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hylander E., Jarnum S., Jensen H. J., Thale M. Enteric hyperoxaluria: dependence on small intestinal resection, colectomy, and steatorrhoea in chronic inflammatory bowel disease. Scand J Gastroenterol. 1978;13(5):577–588. doi: 10.3109/00365527809181767. [DOI] [PubMed] [Google Scholar]
  10. JAKOBY W. B., BHAT J. V. Microbial metabolism of oxalic acid. Bacteriol Rev. 1958 Jun;22(2):75–80. doi: 10.1128/br.22.2.75-80.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. James L. F. Oxalate toxicosis. Clin Toxicol. 1972;5(2):231–243. doi: 10.3109/15563657208991002. [DOI] [PubMed] [Google Scholar]
  12. KORNBERG H. L., ELSDEN S. R. The metabolism of 2-carbon compounds by microorganisms. Adv Enzymol Relat Subj Biochem. 1961;23:401–470. doi: 10.1002/9780470122686.ch8. [DOI] [PubMed] [Google Scholar]
  13. Lung H. Y., Baetz A. L., Peck A. B. Molecular cloning, DNA sequence, and gene expression of the oxalyl-coenzyme A decarboxylase gene, oxc, from the bacterium Oxalobacter formigenes. J Bacteriol. 1994 Apr;176(8):2468–2472. doi: 10.1128/jb.176.8.2468-2472.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Matthews L. A., Doershuk C. F., Stern R. C., Resnick M. I. Urolithiasis and cystic fibrosis. J Urol. 1996 May;155(5):1563–1564. [PubMed] [Google Scholar]
  15. Rodby R. A., Tyszka T. S., Williams J. W. Reversal of cardiac dysfunction secondary to type 1 primary hyperoxaluria after combined liver-kidney transplantation. Am J Med. 1991 Apr;90(4):498–504. [PubMed] [Google Scholar]
  16. Ruan Z. S., Anantharam V., Crawford I. T., Ambudkar S. V., Rhee S. Y., Allison M. J., Maloney P. C. Identification, purification, and reconstitution of OxlT, the oxalate: formate antiport protein of Oxalobacter formigenes. J Biol Chem. 1992 May 25;267(15):10537–10543. [PubMed] [Google Scholar]
  17. SHIMAZONO H., HAYAISHI O. Enzymatic decarboxylation of oxalic acid. J Biol Chem. 1957 Jul;227(1):151–159. [PubMed] [Google Scholar]
  18. Sidhu H., Allison M., Peck A. B. Identification and classification of Oxalobacter formigenes strains by using oligonucleotide probes and primers. J Clin Microbiol. 1997 Feb;35(2):350–353. doi: 10.1128/jcm.35.2.350-353.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Varshavsky A. The N-end rule. Cell. 1992 May 29;69(5):725–735. doi: 10.1016/0092-8674(92)90285-k. [DOI] [PubMed] [Google Scholar]
  20. Williams H. E., Smith L. H., Jr Disorders of oxalate metabolism. Am J Med. 1968 Nov;45(5):715–735. doi: 10.1016/0002-9343(68)90207-6. [DOI] [PubMed] [Google Scholar]
  21. Wyman H., Palmer J. K. Organic Acids in the Ripening Banana Fruit. Plant Physiol. 1964 Jul;39(4):630–633. doi: 10.1104/pp.39.4.630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yanofsky C. Attenuation in the control of expression of bacterial operons. Nature. 1981 Feb 26;289(5800):751–758. doi: 10.1038/289751a0. [DOI] [PubMed] [Google Scholar]

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