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Identification of genes involved in shea butter biosynthesis from Vitellaria paradoxa fruits through transcriptomics and functional heterologous expression

机译:通过转录组学和功能性异源表达鉴定与Vitellaria paradoxa果实乳木果生物合成有关的基因

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摘要

Shea tree (Vitellaria paradoxa) is one economically important plant species that mainly distributes in West Africa. Shea butter extracted from shea fruit kernels can be used as valuable products in the food and cosmetic industries. The most valuable composition in shea butter was one kind of triacylglycerol (TAG), 1,3-distearoyl-2-oleoyl-glycerol (SOS, C18:0–C18:1–C18:0). However, shea butter production is limited and little is known about the genetic information of shea tree. In this study, we tried to reveal genetic information of shea tree and identified shea TAG biosynthetic genes for future shea butter production in yeast cell factories. First, we measured lipid content, lipid composition, and TAG composition of seven shea fruits at different ripe stages. Then, we performed transcriptome analysis on two shea fruits containing obviously different levels of SOS and revealed a list of TAG biosynthetic genes potentially involved in TAG biosynthesis. In total, 4 glycerol-3-phosphate acyltransferase (GPAT) genes, 8 lysophospholipid acyltransferase (LPAT) genes, and 11 diacylglycerol acyltransferase (DGAT) genes in TAG biosynthetic pathway were predicted from the assembled transcriptome and 14 of them were cloned from shea fruit cDNA. Furthermore, the heterologous expression of these 14 potential GPAT, LPAT, and DGAT genes in Saccharomyces cerevisiae changed yeast fatty acid and lipid profiles, suggesting that they functioned in S. cerevisiae. Moreover, two shea DGAT genes, VpDGAT1 and VpDGAT7, were identified as functional DGATs in shea tree, showing they might be useful for shea butter (SOS) production in yeast cell factories.Electronic supplementary materialThe online version of this article (10.1007/s00253-019-09720-3) contains supplementary material, which is available to authorized users.
机译:乳木果(Vitellaria paradoxa)是一种经济上重要的植物,主要​​分布在西非。从乳木果仁中提取的乳木果油可在食品和化妆品工业中用作有价值的产品。乳木果油中最有价值的成分是一种三酰基甘油(TAG),1,3-二硬脂酰基-2-油酰基甘油(SOS,C18:0–C18:1–C18:0)。但是,乳木果油的生产是有限的,对乳木果树的遗传信息知之甚少。在这项研究中,我们试图揭示乳木果的遗传信息,并确定了乳木果TAG生物合成基因,以供将来在酵母细胞工厂中生产乳木果油。首先,我们测量了七个乳木果在不同成熟阶段的脂质含量,脂质组成和TAG组成。然后,我们对含有明显不同SOS水平的两个乳木果进行了转录组分析,并揭示了可能与TAG生物合成有关的TAG生物合成基因的列表。通过组装的转录组预测TAG生物合成途径中共有4个甘油-3-磷酸酰基转移酶(GPAT)基因,8个溶血磷脂酰基转移酶(LPAT)基因和11个二酰基甘油酰基转移酶(DGAT)基因,并且从乳木果中克隆了其中的14个cDNA。此外,在酿酒酵母中这14种潜在的GPAT,LPAT和DGAT基因的异源表达改变了酵母的脂肪酸和脂质谱,表明它们在酿酒酵母中起作用。此外,在乳木果树中鉴定出两个乳木果DGAT基因VpDGAT1和VpDGAT7是功能性DGAT,表明它们可能对酵母细胞工厂中的乳木果油(SOS)生产有用。电子补充材料本文的在线版本(10.1007 / s00253- 019-09720-3)包含补充材料,授权用户可以使用。

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