| Alternative title | Abstracts presented at the 37th Annual Meeting of The Protein Society. ABS#599: Catechol 1,2-dioxygenase with improved thermostability |
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| Author | Search for: Lister, Joshua1; Search for: St-Jacques, Antony1; Search for: Loewen, Michele1 |
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| Affiliation | - National Research Council of Canada. Aquatic and Crop Resource Development
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| Format | Text, Abstract |
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| Conference | 37th Annual Meeting of The Protein Society, July 13-16, 2023, Boston, MA, USA |
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| Abstract | Catechol 1,2-dioxygenase (C12O) is involved in the catabolism of aromatic compounds and biosynthesis of cis, cis-muconic acid, an important precursor chemical and a compound of potential relevance to petrochemical replacement. However, the low thermostability of C12O is a hindrance to industrial application performance. In this study, the thermostability of a mesophilic C12O from Rhodococcus opacus was enhanced by the rational design and engineering of disulphide bonds in the protein structure. A computational pipeline of existing algorithms was used to identify possible locations for the introduction of stabilizing disulphide bonds. A set of selection criteria were used to narrow down the list of candidates, and molecular dynamic simulations were performed for each of the selected candidates under simulated thermal denaturing conditions. Subsequent wet bench testing of the selected candidates showed that two disulfide locations in particular (including an intra-subunit bond and an inter-subunit bond) yielded significant increases to protein thermostability. These two mutants showed greater than 5°C rises in melting temperature and T50, as well as significant increases in half-life at 40°C. Current work on a double mutant of these is ongoing to determine whether the effects of increased stability are additive. Overall, we conclude that the introduction of disulfide bonds resulted in significantly improved thermostability of C12O compared to wild type. |
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| Publication date | 2023-11-27 |
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| Publisher | Wiley |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | e9ad043d-db6a-4953-afec-1c09af2a5482 |
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| Record created | 2024-06-07 |
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| Record modified | 2024-06-07 |
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