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ORIGINAL RESEARCH article

Front. Plant Sci.
Sec. Crop and Product Physiology
Volume 15 - 2024 | doi: 10.3389/fpls.2024.1502454
This article is part of the Research Topic Improving Legume Crops for Resistance to Pathogens and Pests View all articles

Optimizing the Thermostability of Tri-ketone Dioxygenase for Engineering Tolerance to Mesotrione Herbicide in Soybean and Cotton

Provisionally accepted
Stephen Duff Stephen Duff 1,2*Lei Shi Lei Shi 3Danqi Chen Danqi Chen 4Xiaoran Fu Xiaoran Fu 5Mingsheng Peng Mingsheng Peng 2,4Clayton Larue Clayton Larue 2Janice Weihe Janice Weihe 2Jessica Koczan Jessica Koczan 2Brian Krebel Brian Krebel 2Qungang Qi Qungang Qi 2
  • 1 Bayer Crop Science (United States), St. Louis, United States
  • 2 Bayer Crop Sciences (United States), Chesterfield, Missouri, 63017, Missouri, United States
  • 3 Encodia Inc, San Diego, United States
  • 4 Ionova Life Science, Shenzhen, China
  • 5 Bayer CropScience (Belgium), Diegem, Belgium

The final, formatted version of the article will be published soon.

    Optimized triketone dioxygenase (TDO) variants with enhanced temperature stability parameters were engineered to enable robust tri-ketone tolerance in transgenic cotton and soybean crops. This herbicide tolerance trait which can metabolize tri-ketone 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 (~39-40 o C). We designed an optimization scheme using 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 . On the other hand, 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 which had better heat stability in vitro and which efficacies against mesotrione equaling the WT TDO in greenhouse and field tests.

    Keywords: Protein Engineering, triketone dioxygenase, 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors, protein variants, weed management

    Received: 26 Sep 2024; Accepted: 23 Dec 2024.

    Copyright: © 2024 Duff, Shi, Chen, Fu, Peng, Larue, Weihe, Koczan, Krebel and Qi. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Stephen Duff, Bayer Crop Science (United States), St. Louis, United States

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.