Enhancement of enzymatic activity by biomolecular condensates through pH buffering
Abstract Biomolecular condensates can affect enzymatic reactions by locally changing not only concentrations of molecules but also their environment. Since protein conformations can differ between the dense and dilute phase, phase separation can particularly modulate enzymes characterized by a confo...
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| Format: | Article |
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-61013-8 |
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| author | F. Stoffel M. Papp M. Gil-Garcia A. M. Küffner A. I. Benítez-Mateos R. P. B. Jacquat N. Galvanetto L. Faltova P. Arosio |
| author_facet | F. Stoffel M. Papp M. Gil-Garcia A. M. Küffner A. I. Benítez-Mateos R. P. B. Jacquat N. Galvanetto L. Faltova P. Arosio |
| author_sort | F. Stoffel |
| collection | DOAJ |
| description | Abstract Biomolecular condensates can affect enzymatic reactions by locally changing not only concentrations of molecules but also their environment. Since protein conformations can differ between the dense and dilute phase, phase separation can particularly modulate enzymes characterized by a conformation-dependent activity. Here, we generate enzymatic condensates containing a lipase from Bacillus thermocatenulatus, which exhibits an equilibrium between a closed, inactive state, and an open, active conformation. We show that the activity of the enzyme increases inside the dense phase, leading to an enhancement of the overall reaction rate in the phase-separated system. Moreover, we demonstrate that these condensates can generate a more basic environment compared to the surrounding solution, maintaining a high enzymatic activity even in a solution pH interval that would be otherwise less favorable for the lipase. We further show that the formation of two phases with distinct pH values optimizes a cascade reaction involving two enzymes with different optimal pH conditions. Our results demonstrate that, through local pH buffering, biomolecular condensates can expand the optimal pH interval for enzymatic reactions and increase their robustness towards changes in environmental parameters. These findings have implications in biology and biotechnology for biocatalytic engineering, for instance for enabling network reactions with enzymes that require distinct pH values. |
| format | Article |
| id | doaj-art-1d558df60d4e4e9fbf17b3e237ac9f9b |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-1d558df60d4e4e9fbf17b3e237ac9f9b2025-08-20T03:05:06ZengNature PortfolioNature Communications2041-17232025-07-0116111110.1038/s41467-025-61013-8Enhancement of enzymatic activity by biomolecular condensates through pH bufferingF. Stoffel0M. Papp1M. Gil-Garcia2A. M. Küffner3A. I. Benítez-Mateos4R. P. B. Jacquat5N. Galvanetto6L. Faltova7P. Arosio8Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichInstitute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichInstitute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichInstitute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichInstitute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichInstitute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichDepartment of Biochemistry, University of ZurichInstitute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichInstitute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH ZurichAbstract Biomolecular condensates can affect enzymatic reactions by locally changing not only concentrations of molecules but also their environment. Since protein conformations can differ between the dense and dilute phase, phase separation can particularly modulate enzymes characterized by a conformation-dependent activity. Here, we generate enzymatic condensates containing a lipase from Bacillus thermocatenulatus, which exhibits an equilibrium between a closed, inactive state, and an open, active conformation. We show that the activity of the enzyme increases inside the dense phase, leading to an enhancement of the overall reaction rate in the phase-separated system. Moreover, we demonstrate that these condensates can generate a more basic environment compared to the surrounding solution, maintaining a high enzymatic activity even in a solution pH interval that would be otherwise less favorable for the lipase. We further show that the formation of two phases with distinct pH values optimizes a cascade reaction involving two enzymes with different optimal pH conditions. Our results demonstrate that, through local pH buffering, biomolecular condensates can expand the optimal pH interval for enzymatic reactions and increase their robustness towards changes in environmental parameters. These findings have implications in biology and biotechnology for biocatalytic engineering, for instance for enabling network reactions with enzymes that require distinct pH values.https://doi.org/10.1038/s41467-025-61013-8 |
| spellingShingle | F. Stoffel M. Papp M. Gil-Garcia A. M. Küffner A. I. Benítez-Mateos R. P. B. Jacquat N. Galvanetto L. Faltova P. Arosio Enhancement of enzymatic activity by biomolecular condensates through pH buffering Nature Communications |
| title | Enhancement of enzymatic activity by biomolecular condensates through pH buffering |
| title_full | Enhancement of enzymatic activity by biomolecular condensates through pH buffering |
| title_fullStr | Enhancement of enzymatic activity by biomolecular condensates through pH buffering |
| title_full_unstemmed | Enhancement of enzymatic activity by biomolecular condensates through pH buffering |
| title_short | Enhancement of enzymatic activity by biomolecular condensates through pH buffering |
| title_sort | enhancement of enzymatic activity by biomolecular condensates through ph buffering |
| url | https://doi.org/10.1038/s41467-025-61013-8 |
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