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细菌胺单酸酶的异胎表达增强了植物中维生素B1的生物合成

Yi-Hsin Chung¹, Ting-Chieh Chen¹, Wen-Ju Yang¹

¹Institute of Plant and Microbial Biology, Academia Sinica, Taipei, 11529, Taiwan.

The Plant journal : for cell and molecular biology

|November 24, 2023

概括

细菌胺二酸盐 (TDP) 激酶增强了植物中维生素B1的产生. 使用细菌ThiL酶的工程植物直接将胺单酸盐 (TMP) 转化为TDP,提高TDP水平并改善植物健康.

科学领域:

生物化学生物化学
植物科学植物科学
遗传学是一个遗传学.

背景情况:

植物和细菌拥有独特的生化途径来合成二酸胺 (TDP),这是维生素B1的生物活性形式.
植物TDP生物合成涉及将胺单酸盐 (TMP) 转化为胺,然后进行酸化到TDP.
细菌TDP合成利用TMP激酶 (ThiL) 进行直接的TMP酸化到TDP.

研究的目的:

研究细菌TDP生物合成途径在植物中增强维生素B1生产的潜力.
为了确定细菌TMP激酶是否可以拯救缺乏TDP生物合成的植物突变物.
评估细菌ThiL异位表达对TDP和Arabidopsis和大米中总维生素B1积累的影响.

主要方法:

阿拉比多普西斯 (Arabidopsis) 需要胺2 (th2) 突变的特征,表现出性表型.
在Arabidopsis中,大肠杆菌Thil和Thil-GFP的宫外表达,以拯救th2突变的表型.
针对转基因Arabidopsis. 在ThiL-GFP的细胞下定位.
在使用构成性 (UBIQUITIN) 和内特异性 (GLUTELIN1) 促进剂的水中,大肠杆菌Thil的子宫外表达.

主要成果:

大肠杆菌Thil的宫外表达拯救了Arabidopsisth2突变的类表型,证实TDP缺乏是原因.
与野生类型相比,在各种细胞区表达ThiL-GFP的转基因阿拉比多普西斯显示TDP积累增加.
具有pUBI:ThiL的转基因大米系在叶子中积累了更高的TDP和总维生素B1,而pGT1:ThiL系在种子中显示了增强的TDP和维生素B1.
在pGT1:ThiL转基因大米的抛光和未抛光种子中观察到总维生素B1的适度增加 (25-30%).

结论:

细菌TMP激酶 (ThiL) 可以有效地绕过本地植物酸酶步骤,在植物中直接将TMP转化为TDP.
使用细菌ThiL的基因工程是一种可行的策略,可以增强米等作物植物的维生素B1 (TDP) 生物合成.
针对Thil的向表达可以增加特定植物组织中的维生素B1含量,在生物强化中具有潜在的应用.

关键词:

阿拉比多普西斯塔利亚纳.埃舍里希亚大肠杆菌 (Escherichia coli) 是一个大肠杆菌.这是一种Oryza sativa.这里是ThiLL.代谢工程是代谢工程.胺单酸盐激酶是一种胺单酸盐.维生素B1 维生素B1 是一种维生素.

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