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A Multi-Enzyme Cascade Reaction for the Production of 2′3′-cGAMP

Authors :
Martin Becker
Patrick Nikel
Jennifer N. Andexer
Stephan Lütz
Katrin Rosenthal
Source :
Biomolecules, Vol 11, Iss 4, p 590 (2021)
Publication Year :
2021
Publisher :
MDPI AG, 2021.

Abstract

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.

Details

Language :
English
ISSN :
2218273X
Volume :
11
Issue :
4
Database :
Directory of Open Access Journals
Journal :
Biomolecules
Publication Type :
Academic Journal
Accession number :
edsdoj.63e5b9c0ac71406490974cc91c907252
Document Type :
article
Full Text :
https://doi.org/10.3390/biom11040590