Background: Nucleoside 5'-triphosphates (NTPs) play an important role in cells in the transfer of
phosphate groups or bioenergy. In vivo, they are ready to be produced, regenerated and consumed in different
kinds of metabolic pathways, and their concentrations are strictly controlled. NTPs are useful reagents that take
part in many biosynthetic processes. However, NTPs are expensive and unstable, which greatly increases the cost
of the final product if a large amount of NTPs is used directly in biosynthesis. Furthermore, during reactions,
NTPs degrade into NDPs and need to be separated from the reaction mixture, making the operation complicated.
Therefore, NTPs are normally regenerated from NDPs, and only very few NTPs are used in the reaction.
Method: Mechanisms of NTP regeneration were analysed, and their applications in the biosynthesis of nucleotides
and their derivates were described. Basically, NTP regeneration involves isolated enzyme systems and
Result: As one type of cofactor regeneration, NTPs can be effectively regenerated by acetate kinase, pyruvate
kinase, and polyphosphate kinase from acetyl phosphate, phosphoenol pyruvate, and polyphosphate, respectively,
or by whole cells of yeast and Corynebacterium ammoniagenes from simple carbohydrates and phosphate. The
NTP-regeneration method is selected primarily due to the main reaction that it is being coupled with. The cost of
phosphate donors and the convenience of integration with the main process should be considered.
Conclusion: Significant advances have been made when NTP regeneration is coupled with other biosynthetic
processes, especially in the preparation of nucleotides, 2'-deoxynucleotides, sugar-nucleotides and their derivatives.