Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton
Optimized triketone dioxygenase (TDO) variants with enhanced temperature stability parameters were engineered to enable robust triketone tolerance in transgenic cotton and soybean crops. This herbicide tolerance trait, which can metabolize triketone herbicides such as mesotrione and tembotrione, cou...
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| Format: | Article |
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Frontiers Media S.A.
2025-02-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2024.1502454/full |
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| author | Stephen M. G. Duff Stephen M. G. Duff Lei Shi Danqi Chen Xiaoran Fu Mingsheng Peng Clayton T. Larue Janice Weihe Jessica Koczan Brian Krebel Qungang Qi |
| author_facet | Stephen M. G. Duff Stephen M. G. Duff Lei Shi Danqi Chen Xiaoran Fu Mingsheng Peng Clayton T. Larue Janice Weihe Jessica Koczan Brian Krebel Qungang Qi |
| author_sort | Stephen M. G. Duff |
| collection | DOAJ |
| description | Optimized triketone dioxygenase (TDO) variants with enhanced temperature stability parameters were engineered to enable robust triketone tolerance in transgenic cotton and soybean crops. This herbicide tolerance trait, which can metabolize triketone herbicides such as mesotrione and tembotrione, could be useful for weed management systems and provide additional tools for farmers to control weeds. TDO has a low melting point (~39°C–40°C). We designed an optimization scheme using a hypothesis-based rational design to improve the temperature stability of TDO. Temperature stabilization resulted in enzymes with Kcat values less than half of wild-type TDO. The best variant TDO had a Kcat of 1.2 min−1 compared to wild-type TDO, which had a Kcat of 2.7 min−1. However Km values did not change much due to temperature stabilization. Recovery of the Kcat without losing heat stability was the focus of additional optimization. Multiple variants were found that had better heat stability in vitro and efficacies against mesotrione equaling the wild-type (WT) TDO in greenhouse and field tests. |
| format | Article |
| id | doaj-art-814092f8cdf5458e896c96ac1f7235fc |
| institution | Kabale University |
| issn | 1664-462X |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj-art-814092f8cdf5458e896c96ac1f7235fc2025-02-04T10:26:58ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-02-011510.3389/fpls.2024.15024541502454Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cottonStephen M. G. Duff0Stephen M. G. Duff1Lei Shi2Danqi Chen3Xiaoran Fu4Mingsheng Peng5Clayton T. Larue6Janice Weihe7Jessica Koczan8Brian Krebel9Qungang Qi10Bayer Crop Science, St. Louis, MO, United StatesBayer Crop Sciences, Chesterfield, MO, United StatesEncodia Inc., San Diego, CA, United StatesIonova Life Science, Shenzhen, ChinaBayer Crop Science, Diegem, BelgiumBayer Crop Sciences, Chesterfield, MO, United StatesBayer Crop Sciences, Chesterfield, MO, United StatesBayer Crop Sciences, Chesterfield, MO, United StatesBayer Crop Sciences, Chesterfield, MO, United StatesBayer Crop Sciences, Chesterfield, MO, United StatesBayer Crop Sciences, Chesterfield, MO, United StatesOptimized triketone dioxygenase (TDO) variants with enhanced temperature stability parameters were engineered to enable robust triketone tolerance in transgenic cotton and soybean crops. This herbicide tolerance trait, which can metabolize triketone herbicides such as mesotrione and tembotrione, could be useful for weed management systems and provide additional tools for farmers to control weeds. TDO has a low melting point (~39°C–40°C). We designed an optimization scheme using a hypothesis-based rational design to improve the temperature stability of TDO. Temperature stabilization resulted in enzymes with Kcat values less than half of wild-type TDO. The best variant TDO had a Kcat of 1.2 min−1 compared to wild-type TDO, which had a Kcat of 2.7 min−1. However Km values did not change much due to temperature stabilization. Recovery of the Kcat without losing heat stability was the focus of additional optimization. Multiple variants were found that had better heat stability in vitro and efficacies against mesotrione equaling the wild-type (WT) TDO in greenhouse and field tests.https://www.frontiersin.org/articles/10.3389/fpls.2024.1502454/fullprotein engineeringtriketone dioxygenase4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitorsprotein variantsweed management |
| spellingShingle | Stephen M. G. Duff Stephen M. G. Duff Lei Shi Danqi Chen Xiaoran Fu Mingsheng Peng Clayton T. Larue Janice Weihe Jessica Koczan Brian Krebel Qungang Qi Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton Frontiers in Plant Science protein engineering triketone dioxygenase 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors protein variants weed management |
| title | Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton |
| title_full | Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton |
| title_fullStr | Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton |
| title_full_unstemmed | Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton |
| title_short | Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton |
| title_sort | optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton |
| topic | protein engineering triketone dioxygenase 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors protein variants weed management |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2024.1502454/full |
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