Version 1
: Received: 2 August 2024 / Approved: 6 August 2024 / Online: 6 August 2024 (07:03:48 CEST)
How to cite:
Monterrey, D. T.; Azcona, L.; Revuelta, J.; Sánchez-Moreno, I.; García-Junceda, E. Polyphosphate Kinase from Burkholderia cenocepacia, One Enzyme Catalyzing a Two Step Cascade Reaction to Synthesize ATP from AMP. Preprints2024, 2024080379. https://doi.org/10.20944/preprints202408.0379.v1
Monterrey, D. T.; Azcona, L.; Revuelta, J.; Sánchez-Moreno, I.; García-Junceda, E. Polyphosphate Kinase from Burkholderia cenocepacia, One Enzyme Catalyzing a Two Step Cascade Reaction to Synthesize ATP from AMP. Preprints 2024, 2024080379. https://doi.org/10.20944/preprints202408.0379.v1
Monterrey, D. T.; Azcona, L.; Revuelta, J.; Sánchez-Moreno, I.; García-Junceda, E. Polyphosphate Kinase from Burkholderia cenocepacia, One Enzyme Catalyzing a Two Step Cascade Reaction to Synthesize ATP from AMP. Preprints2024, 2024080379. https://doi.org/10.20944/preprints202408.0379.v1
APA Style
Monterrey, D. T., Azcona, L., Revuelta, J., Sánchez-Moreno, I., & García-Junceda, E. (2024). Polyphosphate Kinase from Burkholderia cenocepacia, One Enzyme Catalyzing a Two Step Cascade Reaction to Synthesize ATP from AMP. Preprints. https://doi.org/10.20944/preprints202408.0379.v1
Chicago/Turabian Style
Monterrey, D. T., Israél Sánchez-Moreno and Eduardo García-Junceda. 2024 "Polyphosphate Kinase from Burkholderia cenocepacia, One Enzyme Catalyzing a Two Step Cascade Reaction to Synthesize ATP from AMP" Preprints. https://doi.org/10.20944/preprints202408.0379.v1
Abstract
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.
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
