Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton.

IF 4.1 2区生物学 Q1 PLANT SCIENCES

Frontiers in Plant Science Pub Date : 2025-02-04 eCollection Date: 2024-01-01 DOI:10.3389/fpls.2024.1502454

Stephen M G Duff, Lei Shi, Danqi Chen, Xiaoran Fu, Mingsheng Peng, Clayton T Larue, Janice Weihe, Jessica Koczan, Brian Krebel, Qungang Qi

{"title":"Optimizing the thermostability of triketone dioxygenase for engineering tolerance to mesotrione herbicide in soybean and cotton.","authors":"Stephen M G Duff, Lei Shi, Danqi Chen, Xiaoran Fu, Mingsheng Peng, Clayton T Larue, Janice Weihe, Jessica Koczan, Brian Krebel, Qungang Qi","doi":"10.3389/fpls.2024.1502454","DOIUrl":null,"url":null,"abstract":"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.","PeriodicalId":12632,"journal":{"name":"Frontiers in Plant Science","volume":"15 ","pages":"1502454"},"PeriodicalIF":4.1000,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11833294/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in Plant Science","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3389/fpls.2024.1502454","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/1/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}

引用次数: 0

Abstract

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 K_cat values less than half of wild-type TDO. The best variant TDO had a K_cat of 1.2 min^-1 compared to wild-type TDO, which had a K_cat of 2.7 min^-1. However K_m values did not change much due to temperature stabilization. Recovery of the K_cat 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.

查看原文本刊更多论文

求助全文

约1分钟内获得全文求助全文

来源期刊

Frontiers in Plant Science PLANT SCIENCES-

CiteScore

7.30

自引率

14.30%

发文量

4844

审稿时长

14 weeks

期刊介绍： In an ever changing world, plant science is of the utmost importance for securing the future well-being of humankind. Plants provide oxygen, food, feed, fibers, and building materials. In addition, they are a diverse source of industrial and pharmaceutical chemicals. Plants are centrally important to the health of ecosystems, and their understanding is critical for learning how to manage and maintain a sustainable biosphere. Plant science is extremely interdisciplinary, reaching from agricultural science to paleobotany, and molecular physiology to ecology. It uses the latest developments in computer science, optics, molecular biology and genomics to address challenges in model systems, agricultural crops, and ecosystems. Plant science research inquires into the form, function, development, diversity, reproduction, evolution and uses of both higher and lower plants and their interactions with other organisms throughout the biosphere. Frontiers in Plant Science welcomes outstanding contributions in any field of plant science from basic to applied research, from organismal to molecular studies, from single plant analysis to studies of populations and whole ecosystems, and from molecular to biophysical to computational approaches. Frontiers in Plant Science publishes articles on the most outstanding discoveries across a wide research spectrum of Plant Science. The mission of Frontiers in Plant Science is to bring all relevant Plant Science areas together on a single platform.