The inventive method for evaluating an X protein encoded by an
Lactobacillus fermentum (L. fermentum ) ntd gene in such a way that the
characteristics thereof are modified consists a) in obtaining the
Lactobacillus fermentum (L. fermentum ) ntd gene mutants by random
mutagenesis, b) in transforming cells containing a [P-] phenotype
provided with vectors containing mutated nucleic acids obtained at the
stage a) coding for the thus modified X* proteins, wherein P- means that
said cells are auxotrophic for a substance P produced by the action of X
on a natural substrate S, c) in culturing said cells in a medium
comprising a substrate S*, wherein S* is an analog to the natural
substrate S of the protein X and d) in selecting the cells [P-::X*] which
survived at the stage c) and ijn which the proteins X* are capable of
carrying out the biosynthesis of the product P based on the substrate S*.
The mutated L. fermentum N-desoxyribosyl transferases have an
N-didesoxyribosyl transferase activity, corresponding nucleic acids,
expression vectors, host cells containing said vectors and an application
for the enzymatic synthesis of 2',3'-didesoxynucleosides and
2',3'-didehydro-2',3'-didesoxynucleosides.