工程5-埃诺尔皮鲁维尔什基酸盐-3-酸盐合成酶来自Halomonas sp. 增加草甘耐药性而没有活动罚款
Yunhao He1, Yu Hua2, Xinyi Chen2
1College of Life Science and Technology, Huazhong Agricultural University, No. 1 Shizishan Street, Wuhan 430070, PR China.
Journal of agricultural and food chemistry
|April 19, 2025
概括
研究人员设计了一种更耐草甘的5-enolpyruvylshikimate-3-phosphate合成酶 (EPSPS). 这种增强的酶,HoEPSPS-4M,显示了开发对除草剂草甘耐药作物的潜力.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 植物科学 植物科学
背景情况:
- 草甘除草剂耐药性对于作物分子育种至关重要.
- 5enolpyruvylshikimate-3-phosphate合成酶 (EPSPS) 是草甘的目标.
- 提高草甘耐药性的工程EPSPS是研究的一个关键领域.
研究的目的:
- 为了增强来自 *Halomonas* sp. 的 EPSPS 的草甘抗性. 通过有针对性的进化.
- 描述工程EPSPS变体的生物化学和生物物理特性.
- 评估工程EPSPS在开发抗除草剂作物的潜力.
主要方法:
- 在 *Halomonas* sp. 的定向进化. 这是EPSPS.
- 生物化学分析测量草甘耐药性和催化效率.
- 异热定位热量计 (ITC) 评估草甘的结合亲和力.
- 分子对接和动力学模拟.
- 在 *Nicotiana benthamiana* 中表达工程EPSPS.
主要成果:
- 工程变异*Ho*EPSPS-4M显示了抗草甘的22倍增加和催化效率的2.3倍改善.
- ITC发现,草甘与*Ho*EPSPS-4M.的结合 afinity 减少了 1.7 倍.
- 分子模拟表明,活性位点和远位突变调节酶活性和动态.
- *Ho*EPSPS-4M在*Nicotiana benthamiana*中过度表达时会产生高抗草甘的作用.
结论:
- 定向进化成功增强了*Halomonas* sp.的草甘耐药性和催化效率. 这是EPSPS.
- G112A突变是减少草甘结合的关键,而其他突变优化了酶功能.
- 工程 *Ho*EPSPS-4M 显示了开发抗草甘作物的巨大潜力.
- 这项研究扩大了对除草剂耐药性和蛋白质工程策略的遗传资源.
更多相关视频
09:27Functional Complementation Analysis FCA: A Laboratory Exercise Designed and Implemented to Supplement the Teaching of Biochemical Pathways
Published on: June 24, 2016
17.6K
07:59A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
Published on: October 4, 2019
9.7K
相关概念视频
ATP Synthase: Structure
11.7K
ATP synthase or ATPase is among the most conserved proteins found in bacteria, mammals, and plants. This enzyme can catalyze a forward reaction in response to the electrochemical gradient, producing ATP from ADP and inorganic phosphate. ATP synthase can also work in a reverse direction by hydrolyzing ATP and generating an electrochemical gradient. Different forms of ATP synthases have evolved special features to meet the specific demands of the cell. Based on their specific feature, ATP...
11.7K
ATP Synthase: Mechanism
13.6K
In animals, the mitochondrial F1F0 ATP synthase is the key protein that synthesizes ATP molecules through a complex catalytic mechanism. While the nuclear genome encodes the majority of ATP synthase subunits, the mitochondrial genome encodes some of the enzyme's most critical components. The formation of this multi-subunit enzyme is a complex multi-step process regulated at the level of transcription, translation, and assembly. Defects in one or more of these steps can result in decreased...
13.6K
Catalytically Perfect Enzymes
3.8K
The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
Most enzymes...
Most enzymes...
3.8K
Allosteric Proteins-ATCase
5.6K
Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
5.6K
Photosystems
4.7K
Photosystems are multiprotein complexes that form the functional units of photosynthesis in plants, algae, and cyanobacteria. They are found embedded in the membrane of tiny sac-like structures called thylakoids placed inside the chloroplast.
Functioning of Photosystems
Photosystems contain many pigment molecules, such as chlorophylls and carotenoids, arranged in a particular organization across two domains — the antenna complex and the reaction center. The main aim of the pigment...
Functioning of Photosystems
Photosystems contain many pigment molecules, such as chlorophylls and carotenoids, arranged in a particular organization across two domains — the antenna complex and the reaction center. The main aim of the pigment...
4.7K
The Calvin Cycle
73.0K
Overview
73.0K
