Osmolyte effect on enzymatic stability and reaction equilibrium of formate dehydrogenase

Gajardo-Parra, Nicolás F.; Akrofi-Mantey, Harold; Ascani, Moreno; Cea-Klapp, Esteban; Matias Garrido, José; Sadowski, Gabriele; Held, Christoph

Osmolyte effect on enzymatic stability and reaction equilibrium of formate dehydrogenase

dc.contributor.author	Gajardo-Parra, Nicolás F.
dc.contributor.author	Akrofi-Mantey, Harold
dc.contributor.author	Ascani, Moreno
dc.contributor.author	Cea-Klapp, Esteban
dc.contributor.author	Matias Garrido, José
dc.contributor.author	Sadowski, Gabriele
dc.contributor.author	Held, Christoph
dc.date.accessioned	2023-03-27T11:24:19Z
dc.date.available	2023-03-27T11:24:19Z
dc.date.issued	2022-11-03
dc.description.abstract	Osmolytes are well-known biocatalyst stabilisers as they promote the folded state of proteins, and a stabilised biocatalyst might also improve reaction kinetics. In this work, the influence of four osmolytes (betaine, glycerol, trehalose, and trimethylamine N-oxide) on the activity and stability of Candida bondinii formate dehydrogenase cbFDH was studied experimentally and theoretically. Scanning differential fluorimetric studies were performed to assess the thermal stability of cbFDH, while UV detection was used to reveal changes in cbFDH activity and reaction equilibrium at osmolyte concentrations between 0.25 and 1 mol kg−1. The thermodynamic model ePC-SAFT advanced allowed predicting the effects of osmolyte on the reaction equilibrium by accounting for interactions involving osmolyte, products, substrates, and water. The results show that osmolytes at low concentrations were beneficial for both, thermal stability and cbFDH activity, while keeping the equilibrium yield at high level. Molecular dynamics simulations were used to describe the solvation around the cbFDH surface and the volume exclusion effect, proofing the beneficial effect of the osmolytes on cbFDH activity, especially at low concentrations of trimethylamine N-oxide and betaine. Different mechanisms of stabilisation (dependent on the osmolyte) show the importance of studying solvent–protein dynamics towards the design of optimised biocatalytic processes.	en
dc.identifier.uri	http://hdl.handle.net/2003/41312
dc.identifier.uri	http://dx.doi.org/10.17877/DE290R-23155
dc.language.iso	en	de
dc.relation.ispartofseries	Phys. Chem. Chem. Phys.;24(45)
dc.rights.uri	https://creativecommons.org/licenses/by/3.0/
dc.subject.ddc	660
dc.subject.rswk	Formiate	de
dc.subject.rswk	Dehydrogenasen	de
dc.subject.rswk	Candida boidinii	de
dc.subject.rswk	Betaine	de
dc.subject.rswk	Glycerin	de
dc.subject.rswk	Trehalose	de
dc.subject.rswk	Trimethylaminoxid	de
dc.subject.rswk	Enzymkinetik	de
dc.subject.rswk	Temperaturbeständigkeit	de
dc.subject.rswk	Molekulardynamik	de
dc.title	Osmolyte effect on enzymatic stability and reaction equilibrium of formate dehydrogenase	en
dc.type	Text	de
dc.type.publicationtype	article	de
dcterms.accessRights	open access
eldorado.secondarypublication	true	de
eldorado.secondarypublication.primarycitation	Phys. Chem. Chem. Phys., 2022,24, 27930-27939	de
eldorado.secondarypublication.primaryidentifier	https://doi.org/10.1039/D2CP04011E	de

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