Cell‐Free Biosynthesis of ω‐Hydroxy Acids Boosted by a Synergistic Combination of Alcohol Dehydrogenases

The activity orchestration of an unprecedented cell‐free enzyme system with self‐sufficient cofactor recycling enables the stepwise transformation of aliphatic diols into ω‐hydroxy acids at the expense of molecular oxygen as electron acceptor. The efficiency of the biosynthetic route was maximized w...

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Bibliographic Details
Published in:ChemSusChem 2022-05, Vol.15 (9), p.e202200397-n/a
Main Authors: Velasco‐Lozano, Susana, Santiago‐Arcos, Javier, Grazia Rubanu, Maria, López‐Gallego, Fernando
Format: Article
Language:English
Subjects:
Acids
Alcohol Dehydrogenase - metabolism
Alcohols
Biocatalysis
Biocatalysts
Biosynthesis
dehydrogenase
Dehydrogenases
Diols
Enantiomers
Enzymes
Hydroxy Acids
lactonase multi-enzyme systems
Oxidation-Reduction
Oxygen
ω-hydroxy acid synthesis
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Summary:The activity orchestration of an unprecedented cell‐free enzyme system with self‐sufficient cofactor recycling enables the stepwise transformation of aliphatic diols into ω‐hydroxy acids at the expense of molecular oxygen as electron acceptor. The efficiency of the biosynthetic route was maximized when two compatible alcohol dehydrogenases were selected as specialist biocatalysts for each one of the oxidative steps required for the oxidative lactonization of diols. The cell‐free system reached up to 100 % conversion using 100 mM of linear C5 diols and performed the desymmetrization of prochiral branched diols into the corresponding ω‐hydroxy acids with an exquisite enantioselectivity (ee>99 %). Green metrics demonstrate superior sustainability of this system compared to traditional metal catalysts and even to whole cells for the synthesis of 5‐hydroxypetanoic acid. Finally, the cell‐free system was assembled into a consortium of heterogeneous biocatalysts that allowed the enzyme reutilization. This cascade illustrates the potential of systems biocatalysis to access new heterofunctional molecules such as ω‐hydroxy acids. Who needs cells? We describe the cell‐free biosynthesis of short ω‐hydroxy acids from bio‐based 1,ω‐diols by a multi‐enzyme system combining two specialized alcohol dehydrogenases. This system is proven to transform a wide scope of diols. Finally, we intensify this cell‐free biotransformation by increasing the substrate concentration to 100 mM and immobilizing the system to recycle a consortium of heterogeneous biocatalysts in consecutive operational cycles.
ISSN:1864-5631
1864-564X
DOI:10.1002/cssc.202200397