Optimised Protocol for Drying Aqueous Enzyme Solutions in Organic Solvents – Comparison of Free and Immobilised Candida antarctica Lipase B
Here, we propose an optimised protocol for controlling the initial water activity (aw) in organic reaction mixtures with soluble lipase to ensure reproducible and consistent enzyme activity measurements. Pre-equilibration above saturated salt solutions was used to set aw of the reaction media, where...
| Published in: | ChemCatChem |
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| Main Authors: | , , , |
| Format: | Article in Journal/Newspaper |
| Language: | English |
| Published: |
2023
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| Subjects: | |
| Online Access: | https://orbit.dtu.dk/en/publications/387250a5-88d4-4eb9-a2fa-1929e1bcbdd6 https://doi.org/10.1002/cctc.202201207 https://backend.orbit.dtu.dk/ws/files/304333369/ChemCatChem_2022_Tj_rnelund_Optimised_Protocol_for_Drying_Aqueous_Enzyme_Solutions_in_Organic_Solvents_Comparison_1_.pdf |
| Summary: | Here, we propose an optimised protocol for controlling the initial water activity (aw) in organic reaction mixtures with soluble lipase to ensure reproducible and consistent enzyme activity measurements. Pre-equilibration above saturated salt solutions was used to set aw of the reaction media, where aw was tracked with a hygrometer. Consistent stirring of the suspension and volume correction to compensate for solvent evaporation were found to be critical. The protocol was tested on Candida antarctica lipase B (CALB) in soluble and immobilised forms in three organic solvents (acetonitrile, methyl tert-butyl ether, and hexane) at four different water activities ranging from 0.12 to 0.97. Soluble and immobilised CALB had similar aw-profiles and showed the highest enzyme activity in hexane. The optimised protocol expands the possibility to study enzyme kinetics from immobilised enzymes to soluble enzymes. |
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