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Enhanced Formaldehyde Detoxification by Overexpression of Glutathione-Dependent Formaldehyde Dehydrogenase from Arabidopsis

机译:通过过量表达来增强甲醛的解毒作用 谷胱甘肽依赖性甲醛脱氢酶 拟南芥

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摘要

The ADH2 gene codes for the Arabidopsis glutathione-dependent formaldehyde dehydrogenase (FALDH), an enzyme involved in formaldehyde metabolism in eukaryotes. In the present work, we have investigated the potential role of FALDH in detoxification of exogenous formaldehyde. We have generated a yeast (Saccharomyces cerevisiae) mutant strain (sfa1Δ) by in vivo deletion of the SFA1 gene that codes for the endogenous FALDH. Overexpression of Arabidopsis FALDH in this mutant confers high resistance to formaldehyde added exogenously, which demonstrates the functional conservation of the enzyme through evolution and supports its essential role in formaldehyde metabolism. To investigate the role of the enzyme in plants, we have generated Arabidopsis transgenic lines with modified levels of FALDH. Plants overexpressing the enzyme show a 25% increase in their efficiency to take up exogenous formaldehyde, whereas plants with reduced levels of FALDH (due to either a cosuppression phenotype or to the expression of an antisense construct) show a marked slower rate and reduced ability for formaldehyde detoxification as compared with the wild-type Arabidopsis. These results show that the capacity to take up and detoxify high concentrations of formaldehyde is proportionally related to the FALDH activity in the plant, revealing the essential role of this enzyme in formaldehyde detoxification.
机译:ADH2基因编码拟南芥谷胱甘肽依赖性甲醛脱氢酶(FALDH),一种涉及真核生物甲醛代谢的酶。在目前的工作中,我们研究了FALDH在外源甲醛解毒中的潜在作用。我们已经通过体内缺失编码内源性FALDH的SFA1基因产生了酵母(Saccharomyces cerevisiae)突变株(sfa1Δ)。在该突变体中拟南芥FALDH的过表达赋予了对外源添加的甲醛的高抗性,这证明了酶通过进化的功能保守性并支持其在甲醛代谢中的重要作用。为了研究该酶在植物中的作用,我们已经产生了具有修饰水平的FALDH的拟南芥转基因品系。过表达该酶的植物吸收外源甲醛的效率提高了25%,而FALDH含量降低(由于共抑制表型或反义构建体的表达)的植物显示出明显的减慢速度,降低了其能力。甲醛解毒与野生型拟南芥相比。这些结果表明,吸收和排毒的能力很高。 甲醛浓度与FALDH活性成正比关系 在植物中,揭示了这种酶在甲醛中的重要作用 排毒。

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