A Multi-Enzyme Cascade Reaction for the Production of 2'3'-cGAMP
Multi-enzyme cascade reactions for the synthesis of complex products have gained importance in recent decades. Their advantages compared to single biotransformations include the possibility to synthesize complex molecules without purification of reaction intermediates, easier handling of unstable intermediates, and dealing with unfavorable thermodynamics by coupled equilibria. In this study, a four-enzyme cascade consisting of ScADK, AjPPK2, and SmPPK2 for ATP synthesis from adenosine coupled to the cyclic GMP-AMP synthase (cGAS) catalyzing cyclic GMP-AMP (2'3'-cGAMP) formation was successfully developed. The 2'3'-cGAMP synthesis rates were comparable to the maximal reaction rate achieved in single-step reactions. An iterative optimization of substrate, cofactor, and enzyme concentrations led to an overall yield of 0.08 mole 2'3'-cGAMP per mole adenosine, which is comparable to chemical synthesis. The established enzyme cascade enabled the synthesis of 2'3'-cGAMP from GTP and inexpensive adenosine as well as polyphosphate in a biocatalytic one-pot reaction, demonstrating the performance capabilities of multi-enzyme cascades for the synthesis of pharmaceutically relevant products.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:11 |
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| Enthalten in: |
Biomolecules - 11(2021), 4 vom: 16. Apr. |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Becker, Martin [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 20.09.2021 Date Revised 01.04.2024 published: Electronic Citation Status MEDLINE |
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| doi: |
10.3390/biom11040590 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 100 | 1 | |a Becker, Martin |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A Multi-Enzyme Cascade Reaction for the Production of 2'3'-cGAMP |
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| 500 | |a published: Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Multi-enzyme cascade reactions for the synthesis of complex products have gained importance in recent decades. Their advantages compared to single biotransformations include the possibility to synthesize complex molecules without purification of reaction intermediates, easier handling of unstable intermediates, and dealing with unfavorable thermodynamics by coupled equilibria. In this study, a four-enzyme cascade consisting of ScADK, AjPPK2, and SmPPK2 for ATP synthesis from adenosine coupled to the cyclic GMP-AMP synthase (cGAS) catalyzing cyclic GMP-AMP (2'3'-cGAMP) formation was successfully developed. The 2'3'-cGAMP synthesis rates were comparable to the maximal reaction rate achieved in single-step reactions. An iterative optimization of substrate, cofactor, and enzyme concentrations led to an overall yield of 0.08 mole 2'3'-cGAMP per mole adenosine, which is comparable to chemical synthesis. The established enzyme cascade enabled the synthesis of 2'3'-cGAMP from GTP and inexpensive adenosine as well as polyphosphate in a biocatalytic one-pot reaction, demonstrating the performance capabilities of multi-enzyme cascades for the synthesis of pharmaceutically relevant products | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a 2′3′-cGAMP | |
| 650 | 4 | |a ATP | |
| 650 | 4 | |a biocatalysis | |
| 650 | 4 | |a cGAS | |
| 650 | 4 | |a cyclic dinucleotide (CDN) | |
| 650 | 4 | |a enzyme cascade | |
| 650 | 4 | |a in vitro biotransformation | |
| 650 | 4 | |a multi-step reaction | |
| 650 | 4 | |a polyphosphate kinase | |
| 650 | 7 | |a Adenine Nucleotides |2 NLM | |
| 650 | 7 | |a Bacterial Proteins |2 NLM | |
| 650 | 7 | |a Nucleotides, Cyclic |2 NLM | |
| 650 | 7 | |a Saccharomyces cerevisiae Proteins |2 NLM | |
| 650 | 7 | |a cyclic guanosine monophosphate-adenosine monophosphate |2 NLM | |
| 650 | 7 | |a Adenosine Kinase |2 NLM | |
| 650 | 7 | |a EC 2.7.1.20 |2 NLM | |
| 650 | 7 | |a Phosphotransferases (Phosphate Group Acceptor) |2 NLM | |
| 650 | 7 | |a EC 2.7.4.- |2 NLM | |
| 650 | 7 | |a polyphosphate kinase |2 NLM | |
| 650 | 7 | |a EC 2.7.4.1 |2 NLM | |
| 700 | 1 | |a Nikel, Patrick |e verfasserin |4 aut | |
| 700 | 1 | |a Andexer, Jennifer N |e verfasserin |4 aut | |
| 700 | 1 | |a Lütz, Stephan |e verfasserin |4 aut | |
| 700 | 1 | |a Rosenthal, Katrin |e verfasserin |4 aut | |
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| 856 | 4 | 0 | |u http://dx.doi.org/10.3390/biom11040590 |3 Volltext |
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