Enzymatic synthesis of carba-nad

Abstract

The disclosure concerns the enzymatic synthesis of stable analogues of nicotinamide adenine dinucleotide NAD/NADH and nicotinamide adenine dinucleotide phosphate NADP/NADPH, the so-called “carba-NADs”, i.e. analogues of NAD/NADH or NADP/NADPH, respectively, comprising a carbacyclic sugar instead of ribose.

What is claimed is: 
     
         1 . A compound, comprising Formula A: 
       
         
           
           
               
               
           
         
         wherein R1 is selected from OH, NH 2 , O-methyl, and N-dimethyl, methyl; R2 comprises one of NH 2 , OH, and NHalkyl; R3 is selected from H, OH and NH 2 ; Y −  comprises a counter ion, and X comprises one of O and S, wherein said compound of Formula A is comprised primarily of the biologically active form of the molecule and is produced by the following steps: 
         a) phosphorylating a compound of Formula I: 
       
       
         
           
           
               
               
           
         
         in a first reaction catalyzed by a nicotinamide-ribosylkinase; 
         b) adenylating the phosphorylated product of step (a) with a compound of Formula II: 
       
       
         
           
           
               
               
           
         
         in a second reaction catalyzed by a nicotinamide mononucleotide-adenylyltransferase. 
       
     
     
         2 . The compound of  claim 1 , wherein the nicotinamide-ribosylkinase is present in at least one  Streptococcus sanguinius  and  Homo sapiens.    
     
     
         3 . The compound of  claim 1 , wherein the nicotinamide mononucleotide-adenylyltransferase is present in at least one of  Saccharomyces cerevisiae  and  Homo sapiens.    
     
     
         4 . The compound of  claim 1 , wherein the nicotinamide mononucleotide-adenylyltransferase is recombinantly generated in at least one of  E. coli  and  B. subtilis.    
     
     
         5 . The compound of  claim 1 , wherein R1 is NH 2 , R2 is NH 2 , R3 is H and X is O .