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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 in...

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Autores principales: Becker, Martin, Nikel, Patrick, Andexer, Jennifer N., Lütz, Stephan, Rosenthal, Katrin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073963/
https://www.ncbi.nlm.nih.gov/pubmed/33923845
http://dx.doi.org/10.3390/biom11040590
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author Becker, Martin
Nikel, Patrick
Andexer, Jennifer N.
Lütz, Stephan
Rosenthal, Katrin
author_facet Becker, Martin
Nikel, Patrick
Andexer, Jennifer N.
Lütz, Stephan
Rosenthal, Katrin
author_sort Becker, Martin
collection PubMed
description 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.
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spelling pubmed-80739632021-04-27 A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP Becker, Martin Nikel, Patrick Andexer, Jennifer N. Lütz, Stephan Rosenthal, Katrin Biomolecules Article 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. MDPI 2021-04-16 /pmc/articles/PMC8073963/ /pubmed/33923845 http://dx.doi.org/10.3390/biom11040590 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Becker, Martin
Nikel, Patrick
Andexer, Jennifer N.
Lütz, Stephan
Rosenthal, Katrin
A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP
title A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP
title_full A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP
title_fullStr A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP
title_full_unstemmed A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP
title_short A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP
title_sort multi-enzyme cascade reaction for the production of 2′3′-cgamp
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073963/
https://www.ncbi.nlm.nih.gov/pubmed/33923845
http://dx.doi.org/10.3390/biom11040590
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