Heterologous Expression of SiFBP, a Folate-Binding Protein from Foxtail Millet, Confers Increased Folate Content and Altered Amino Acid Profiles with Nutritional Potential to Arabidopsis,Journal of Agricultural and Food Chemistry

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Heterologous Expression of SiFBP, a Folate-Binding Protein from Foxtail Millet, Confers Increased Folate Content and Altered Amino Acid Profiles with Nutritional Potential to Arabidopsis
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-05-16 , DOI: 10.1021/acs.jafc.2c00357
Siyu Hou _{1,

2} , Yijuan Zhang _{1,

2} , Bing Zhao ₃ , Xiaxia Man ₁ , Guifang Ma ₁ , Yihan Men ₁ , Wei Du ₁ , Yang Yang _{1,

2} , Hongying Li _{1,

2} , Yuanhuai Han _{1,

2} , Yaofei Zhao _{1,

2} , Zhaoxia Sun _{1,

2}

Affiliation

The mechanism underlying folate degradation in foxtail millet grains remains unclear. Here, we identified SiFBP (Setaria italica folate-binding protein) from foxtail millet. A phylogenetic tree revealed that FBPs have close genetic relationships among cereal crop species. Docking analysis and heterologous expression of SiFBP in yeast showed that it could bind folic acid (FA). The SiFBP localized to the plasma membrane in tobacco mesophyll cells by transient expression. In Arabidopsis, it was expressed specifically in the roots and germinating seeds. Overexpressing SiFBP in yeast and Arabidopsis significantly increased folate contents. Untargeted metabolome analysis revealed differentially accumulated metabolites between the transgenic lines (TLs) and wild type (WT); these metabolites were mainly enriched in the amino acid metabolism pathway. The relative contents of lysine and leucine, threonine, and l-methionine were significantly higher in the TLs than in WT. Genes related to the folate and lysine synthesis pathways were upregulated in the TLs. Thus, SiFBP can be used for biofortification of folate and important amino acids in crops via genetic engineering.

中文翻译：

SiFBP（一种来自谷子的叶酸结合蛋白）的异源表达赋予拟南芥增加叶酸含量和改变氨基酸谱的营养潜力

谷子中叶酸降解的机制仍不清楚。在这里，我们从谷子中鉴定出SiFBP（Setaria italica叶酸结合蛋白）。系统发育树显示，FBPs 在谷类作物物种之间具有密切的遗传关系。SiFBP在酵母中的对接分析和异源表达表明它可以结合叶酸（FA）。SiFBP通过瞬时表达定位于烟草叶肉细胞中的质膜。在拟南芥中，它在根和发芽的种子中特异性表达。在酵母和拟南芥中过表达SiFBP叶酸含量显着增加。非靶向代谢组分析揭示了转基因系 (TL) 和野生型 (WT) 之间差异积累的代谢物；这些代谢物主要富集在氨基酸代谢途径中。TL中赖氨酸和亮氨酸、苏氨酸和L-蛋氨酸的相对含量显着高于WT。与叶酸和赖氨酸合成途径相关的基因在 TL 中上调。因此，SiFBP可用于通过基因工程对作物中的叶酸和重要氨基酸进行生物强化。

更新日期：2022-05-16

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