Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactions
Polyphosphate kinases (PPKs) have become popular biocatalysts for nucleotide 5'-triphosphate (NTP) synthesis and regeneration. Two unrelated families are described: PPK1 and PPK2. They are structurally unrelated and use different catalytic mechanisms. PPK1 enzymes prefer the usage of adenosine...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Beilstein-Institut
2022-09-01
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| Series: | Beilstein Journal of Organic Chemistry |
| Subjects: | |
| Online Access: | https://doi.org/10.3762/bjoc.18.134 |
| _version_ | 1817980319227183104 |
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| author | Michael Keppler Sandra Moser Henning J. Jessen Christoph Held Jennifer N. Andexer |
| author_facet | Michael Keppler Sandra Moser Henning J. Jessen Christoph Held Jennifer N. Andexer |
| author_sort | Michael Keppler |
| collection | DOAJ |
| description | Polyphosphate kinases (PPKs) have become popular biocatalysts for nucleotide 5'-triphosphate (NTP) synthesis and regeneration. Two unrelated families are described: PPK1 and PPK2. They are structurally unrelated and use different catalytic mechanisms. PPK1 enzymes prefer the usage of adenosine 5'-triphosphate (ATP) for polyphosphate (polyP) synthesis while PPK2 enzymes favour the reverse reaction. With the emerging use of PPK enzymes in biosynthesis, a deeper understanding of the enzymes and their thermodynamic reaction course is of need, especially in comparison to other kinases. Here, we tested four PPKs from different organisms under the same conditions without any coupling reactions. In comparison to other kinases using phosphate donors with comparably higher phosphate transfer potentials that are characterised by reaction yields close to full conversion, the PPK-catalysed reaction reaches an equilibrium in which about 30% ADP is left. These results were obtained for PPK1 and PPK2 enzymes, and are supported by theoretical data on the basic reaction. At high concentrations of substrate, the different kinetic preferences of PPK1 and PPK2 can be observed. The implications of these results for the application of PPKs in chemical synthesis and as enzymes for ATP regeneration systems are discussed. |
| first_indexed | 2024-04-13T22:52:41Z |
| format | Article |
| id | doaj.art-ef97fa7dfa054971945ad63cb6f0a6b2 |
| institution | Directory Open Access Journal |
| issn | 1860-5397 |
| language | English |
| last_indexed | 2024-04-13T22:52:41Z |
| publishDate | 2022-09-01 |
| publisher | Beilstein-Institut |
| record_format | Article |
| series | Beilstein Journal of Organic Chemistry |
| spelling | doaj.art-ef97fa7dfa054971945ad63cb6f0a6b22022-12-22T02:26:08ZengBeilstein-InstitutBeilstein Journal of Organic Chemistry1860-53972022-09-011811278128810.3762/bjoc.18.1341860-5397-18-134Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactionsMichael Keppler0Sandra Moser1Henning J. Jessen2Christoph Held3Jennifer N. Andexer4Institute of Pharmaceutical Sciences, University of Freiburg, Albertstr. 25, 79104 Freiburg, Germany Institute of Organic Chemistry, University of Freiburg, Albertstr. 21, 79104 Freiburg, Germany Institute of Organic Chemistry, University of Freiburg, Albertstr. 21, 79104 Freiburg, Germany Department of Biochemical and Chemical Engineering, TU Dortmund University, Emil-Figge-Str. 70, 44227 Dortmund, Germany Institute of Pharmaceutical Sciences, University of Freiburg, Albertstr. 25, 79104 Freiburg, Germany Polyphosphate kinases (PPKs) have become popular biocatalysts for nucleotide 5'-triphosphate (NTP) synthesis and regeneration. Two unrelated families are described: PPK1 and PPK2. They are structurally unrelated and use different catalytic mechanisms. PPK1 enzymes prefer the usage of adenosine 5'-triphosphate (ATP) for polyphosphate (polyP) synthesis while PPK2 enzymes favour the reverse reaction. With the emerging use of PPK enzymes in biosynthesis, a deeper understanding of the enzymes and their thermodynamic reaction course is of need, especially in comparison to other kinases. Here, we tested four PPKs from different organisms under the same conditions without any coupling reactions. In comparison to other kinases using phosphate donors with comparably higher phosphate transfer potentials that are characterised by reaction yields close to full conversion, the PPK-catalysed reaction reaches an equilibrium in which about 30% ADP is left. These results were obtained for PPK1 and PPK2 enzymes, and are supported by theoretical data on the basic reaction. At high concentrations of substrate, the different kinetic preferences of PPK1 and PPK2 can be observed. The implications of these results for the application of PPKs in chemical synthesis and as enzymes for ATP regeneration systems are discussed.https://doi.org/10.3762/bjoc.18.134atp regenerationbiocatalystepc-saftpolypppk |
| spellingShingle | Michael Keppler Sandra Moser Henning J. Jessen Christoph Held Jennifer N. Andexer Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactions Beilstein Journal of Organic Chemistry atp regeneration biocatalyst epc-saft polyp ppk |
| title | Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactions |
| title_full | Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactions |
| title_fullStr | Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactions |
| title_full_unstemmed | Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactions |
| title_short | Make or break: the thermodynamic equilibrium of polyphosphate kinase-catalysed reactions |
| title_sort | make or break the thermodynamic equilibrium of polyphosphate kinase catalysed reactions |
| topic | atp regeneration biocatalyst epc-saft polyp ppk |
| url | https://doi.org/10.3762/bjoc.18.134 |
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