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Biosynthesis of Phytosterol Esters: Identification of a Sterol O-Acyltransferase in Arabidopsis

机译:植物甾醇酯的生物合成:拟南芥中甾醇O-酰基转移酶的鉴定

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摘要

Fatty acyl esters of phytosterols are a major form of sterol conjugates distributed in many parts of plants. In this study we report an Arabidopsis (Arabidopsis thaliana) gene, AtSAT1 (At3g51970), which encodes for a novel sterol O-acyltransferase. When expressed in yeast (Saccharomyces cerevisiae), AtSAT1 mediated production of sterol esters enriched with lanosterol. Enzyme property assessment using cell-free lysate of yeast expressing AtSAT1 suggested the enzyme preferred cycloartenol as acyl acceptor and saturated fatty acyl-Coenyzme A as acyl donor. Taking a transgenic approach, we showed that Arabidopsis seeds overexpressing AtSAT1 accumulated fatty acyl esters of cycloartenol, accompanied by substantial decreases in ester content of campesterol and β-sitosterol. Furthermore, fatty acid components of sterol esters from the transgenic lines were enriched with saturated and long-chain fatty acids. The enhanced AtSAT1 expression resulted in decreased level of free sterols, but the total sterol content in the transgenic seeds increased by up to 60% compared to that in wild type. We conclude that AtSAT1 mediates phytosterol ester biosynthesis, alternative to the route previously described for phospholipid:sterol acyltransferase, and provides the molecular basis for modification of phytosterol ester level in seeds.
机译:植物甾醇的脂肪酰基酯是分布在植物许多部位的甾醇结合物的主要形式。在这项研究中,我们报告了拟南芥(Arabidopsis thaliana)基因AtSAT1(At3g51970),该基因编码一种新型固醇O-酰基转移酶。当在酵母(酿酒酵母)中表达时,AtSAT1介导富含羊毛甾醇的固醇酯的产生。使用表达AtSAT1的酵母的无细胞裂解物进行的酶特性评估表明,该酶优选环戊烯醇作为酰基受体,饱和脂肪酰基辅酶A作为酰基供体。采取转基因方法,我们显示了过表达AtSAT1的拟南芥种子积累了环烯醇的脂肪酰基酯,同时菜油甾醇和β-谷甾醇的酯含量大大降低。此外,来自转基因品系的固醇酯的脂肪酸成分富含饱和和长链脂肪酸。增强的AtSAT1表达导致游离固醇含量降低,但与野生型相比,转基因种子中的总固醇含量最多增加60%。我们得出的结论是,AtSAT1介导了植物甾醇酯的生物合成,替代了先前描述的磷脂:甾醇酰基转移酶的途径,并为修饰种子中植物甾醇酯的水平提供了分子基础。

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