Materials and methods for detection of oxalate

Abstract

The subject invention concerns the novel use of formyl-CoA transferase enzyme together with oxalyl-CoA decarboxylase enzyme for the detection and measurement of oxalate in biological samples. The use of the enzyme system according to the subject invention results in the conversion of oxalate into carbon dioxide and formate. Because the production of formate is directly correlated to the concentration of oxalate present in a sample, the determination of the resulting formate concentration provides an accurate, sensitive and rapid means for detecting even low levels of oxalate. The subject invention further concerns the cloning, sequencing and expression of the genes that encode the formyl-CoA transferase enzyme and the oxalyl-CoA decarboxylase enzyme of Oxalobacter formigenes.

I claim:  
     
         1 . A purified polynucleotide molecule, consisting of a nucleotide sequence that encodes oxalyl-CoA decarboxylase comprising the amino acid sequence shown in SEQ ID NO. 7, or an enzymatically active fragment of said oxalyl-CoA decarboxylase.  
     
     
         2 . The polynucleotide according to    claim 1   , wherein said polynucleotide hybridizes with a nucleotide sequence comprising nucleotides  181  through  1884  of the nucleotide sequence shown in SEQ ID NO. 6 or the complementary sequence thereof.  
     
     
         3 . The polynucleotide according to    claim 1   , wherein said polynucleotide has a nucleotide sequence comprising nucleotides  181  through  1884  of the nucleotide sequence shown in SEQ ID NO. 6 or the complementary sequence thereof.  
     
     
         4 . A kit comprising in one or more containers a polynucleotide of    claim 1   .  
     
     
         5 . A recombinant host cell which comprises the polynucleotide of    claim 1   , and wherein said cell expresses said oxalyl-CoA decarboxylase or an enzymatically active fragment of said oxalyl-CoA decarboxylase.