ATP is an essential molecule in many biocatalytic processes. The industrial implementation of these processes is hampered by the fact that they require stoichiometric quantities of this expen-sive co-factor. To avoid this problem, recycling systems are needed. This study characterizes a putative polyphosphate kinase enzyme from Burkholderia cenocepacia that can regenerate ATP from AMP. Sequence analysis shows that BcPPK2-III exhibits the characteristic structural motifs of known PPK2 enzymes, including conserved motifs Walker A and B, and the subclass-specific res-idue E137. Molecular docking simulations showed ADP had the highest binding affinity (-7.340 kcal/mol), followed by AMP (-7.284 kcal/mol), with ATP having the lowest affinity (-6.464 kcal/mol). The enzyme was overexpressed in E. coli. Protein expression and purification were monitored by SDS-PAGE and quantified by densitometric analysis. Enzymatic activity assays re-vealed that BcPPK2-III converts 70% ATP with a TTN of 334,000 after 24 hours and remains sta-ble over a pH range of 6-9 and temperatures up to 70°C. BcPPK2-III shows significant potential for industrial ATP regeneration and offers high conversion rates and stability. The enzyme's high conversion rate and stability make BcPPK2-III a promising candidate for the improvement of bio-catalytic processes that rely on ATP at industrial level.