Stabilization of pyruvate decarboxylase under a pressurized carbon dioxide/water biphasic system

Matsuda, Tomoko; Nakayama, Koji; Abe, Taisuke; Mukouyama, Masaharu

doi:10.3109/10242421003734696

Cited by 7 publications

(2 citation statements)

References 16 publications

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“…To the best of our knowledge, there is no experimental or theoretical work in which the effects of natural osmolytes on the structure of a protein in a supercritical fluid have been investigated. In the only rare work, it was experimentally observed by Matsuda et al 58 that pyruvate decarboxylase loses 80% of its activity in a pressurized CO 2 /water biphasic system, but upon addition of trehalose, activity was improved significantly. Although their system was not neat or used under supercritical conditions, their obtained experimental data can confirm our results about the scCO 2 /trehalose system.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO₂: Molecular Level Evidence

2015

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Protein instability in supercritical CO2 limits the application of this green solvent in enzyme-catalyzed reactions. CO2 molecules act as a protein denaturant at high pressure under supercritical conditions. Here, for the first time, we show that natural osmolytes could stabilize protein conformation in supercritical CO2. Molecular dynamics simulation is used to monitor the effects of adding different natural osmolytes on the conformation and dynamics of chymotrypsin inhibitor 2 (CI2) in supercritical CO2. Simulations showed that CI2 is denatured at 200 bar in supercritical CO2, which is in agreement with experimental observations. Interestingly, the protein conformation remains native after addition of ∼1 M amino acid- and sugar-based osmolyte models. These molecules stabilize protein through the formation of supramolecular self-assemblies resulting from macromolecule-osmolyte hydrogen bonds. Nevertheless, trimethylamine N-oxide, which is known as a potent osmolyte for protein stabilization in aqueous solutions, amplifies protein denaturation in supercritical CO2. On the basis of our structural analysis, we introduce a new mechanism for the osmolyte effect in supercritical CO2, an "inclusion mechanism". To the best of our knowledge, this is the first study that introduces the application of natural osmolytes in a supercritical fluid and describes mechanistic insights into osmolyte action in nonaqueous media.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO₂: Molecular Level Evidence

2015

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“…Optimization of reaction conditions to enhance catalyst stability and allow scCO 2 exploitation as a suitable medium for enzymatic carboxylation are key. One study shows that pyruvate decarboxylase loses 80 % of its decarboxylation activity following treatment with pressurized CO 2 at 60 bar, [92] with enzyme inactivation mitigated through the use of excipients such as glycerol and trehalose, and immobilization on an ion exchange polymer. These interventions led to a dramatic improvement in the biocatalyst stability.…”

Section: Strategies To Improve Carboxylation Yieldsmentioning

confidence: 99%

Synthetic Enzyme‐Catalyzed CO₂ Fixation Reactions

et al. 2021

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In recent years, (de)carboxylases that catalyze reversible (de)carboxylation have been targeted for application as carboxylation catalysts. This has led to the development of proof‐of‐concept (bio)synthetic CO2 fixation routes for chemical production. However, further progress towards industrial application has been hampered by the thermodynamic constraint that accompanies fixing CO2 to organic molecules. In this Review, biocatalytic carboxylation methods are discussed with emphases on the diverse strategies devised to alleviate the inherent thermodynamic constraints and their application in synthetic CO2‐fixation cascades.

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Biocatalysis in Organic Solvents, Supercritical Fluids and Ionic Liquids

Cao¹,

Matsuda²

2016

Organic Synthesis Using Biocatalysis

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Stabilization of pyruvate decarboxylase under a pressurized carbon dioxide/water biphasic system

Cited by 7 publications

References 16 publications

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO₂: Molecular Level Evidence

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO₂: Molecular Level Evidence

Synthetic Enzyme‐Catalyzed CO₂ Fixation Reactions

Biocatalysis in Organic Solvents, Supercritical Fluids and Ionic Liquids

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Stabilization of pyruvate decarboxylase under a pressurized carbon dioxide/water biphasic system

Cited by 7 publications

References 16 publications

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO2: Molecular Level Evidence

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO2: Molecular Level Evidence

Synthetic Enzyme‐Catalyzed CO2 Fixation Reactions

Biocatalysis in Organic Solvents, Supercritical Fluids and Ionic Liquids

Contact Info

Product

Resources

About

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO₂: Molecular Level Evidence

Effects of Natural Osmolytes on the Protein Structure in Supercritical CO₂: Molecular Level Evidence

Synthetic Enzyme‐Catalyzed CO₂ Fixation Reactions