Bacteria engineered to treat disorders in which oxalate is detrimental

Abstract

The present invention provides recombinant bacterial cells comprising at least one heterologous gene encoding at least one oxalate catabolism enzyme. In another aspect, the recombinant bacterial cells further comprise at least one heterologous gene encoding an importer of oxalate. The invention further provides pharmaceutical compositions comprising the recombinant bacteria, and methods for treating disorders in which oxalate is detrimental, such as hyperoxaluria, using the pharmaceutical compositions of the invention.

1 .- 63 . (canceled) 
     
     
         64 . A bacterium comprising gene sequences encoding one or more oxalate catabolism enzymes operably linked to a first promoter that is not associated with the oxalate catabolism enzyme gene in nature, wherein the gene sequences encoding the one or more oxalate catabolism enzymes encode ScAAE3, Oxc, and Frc. 
     
     
         65 . The bacterium of  claim 64 , wherein the bacterium further comprises a gene sequence encoding a transporter of oxalate operably linked to a second promoter that is not associated with the transporter gene sequence in nature. 
     
     
         66 . The bacterium of  claim 65 , wherein the bacterium further comprises a gene sequences encoding an exporter of formate operably linked to a third promoter that is not associated with the exporter gene sequence in nature. 
     
     
         67 . The bacterium of  claim 64 , wherein the bacterium further comprises a knockout of a clbA gene. 
     
     
         68 . The bacterium of  claim 64 , wherein the bacterial cell is an auxotroph in a gene that is complemented when the bacterial cell is present in a mammalian gut. 
     
     
         69 . The bacterium of  claim 68 , wherein the bacterial cell is an auxotroph in diaminopimelic acid or an enzyme in the thymine biosynthetic pathway. 
     
     
         70 . The bacterium of  claim 64 , wherein the one or more oxalate catabolism enzymes convert oxalate to formate. 
     
     
         71 . The bacterium of  claim 64 , wherein the bacterium is a probiotic bacterial cell. 
     
     
         72 . The bacterium of  claim 71 , wherein the bacterium is a member of a genus selected from the group consisting of  Bacteroides, Bifidobacterium, Clostridium, Escherichia, Lactobacillus  and  Lactococcus.    
     
     
         73 . A pharmaceutical composition comprising the bacterium of  claim 64 , and a pharmaceutically acceptable carrier. 
     
     
         74 . A method for reducing the levels of oxalate in a subject, the method comprising administering a pharmaceutical composition of  claim 73  to the subject. 
     
     
         75 . A method for treating a disease or disorder in which oxalate is detrimental in a subject, the method comprising administering a pharmaceutical composition of  claim 73 . 
     
     
         76 . The method of  claim 75 , wherein the disorder in which oxalate is detrimental is a hyperoxaluria.