Deletion of Plasma Membrane Malate Transporters Increased Lipid Accumulation in the Oleaginous Fungus Mucor circinelloides WJ11
Deletion of Plasma Membrane Malate Transporters Increased Lipid Accumulation in the Oleaginous Fungus Mucor circinelloides WJ11
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DOI：
10.1021/acs.jafc.1c03307
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发表时间：
2021-08-16
影响因子：
6.1
通讯作者：
Song, Yuanda
中科院分区：
文献类型：
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作者：
Yang, Junhuan;Canovas-Marquez, Jose T.;Song, Yuanda
苹果酸盐作为一种重要的中间代谢物，其亚细胞位置和浓度对真菌脂质代谢有显著影响。先前的研究表明，线粒体苹果酸转运体通过调控细胞内苹果酸浓度在毛霉环状毛霉脂质积累中起重要作用。然而，质膜苹果酸转运体在产油真菌中的作用仍未被探索。因此，本研究删除了M. circinelloides WJ11的两个质膜苹果酸转运体“2- oxogglutarate：苹果酸反转运体”（命名为SoDIT-a和SoDIT-b），并分析了其对生长能力、脂质积累和代谢的影响。结果表明，sodit-a或/和sodit-b的缺失减少了细胞外苹果酸，证实了这两个基因的产物参与了苹果酸运输。与此同时，突变体的脂质含量比对照菌株增加了约10-40%，这表明质膜苹果酸转运缺陷导致可用于脂质生物合成的苹果酸盐增加。此外，转录分析显示，敲除突变体中涉及脂质生物合成的多个关键基因的表达水平也有所增加。据我们所知，这是第一次报道证明了质膜苹果酸转运蛋白和圆叶线虫脂质积累之间的关系。
Malate as an important intermediate metabolite, its subcellular location, and concentration have a significant impact on fungal lipid metabolism. Previous studies showed that the mitochondrial malate transporter plays an important role in lipid accumulation in Mucor circinelloides by manipulating intracellular malate concentration. However, the role of plasma membrane malate transporters in oleaginous fungi remains unexplored. Therefore, in this work, two plasma membrane malate transporters "2-oxoglutarate:malate antiporters" (named SoDIT-a and SoDIT-b) of M. circinelloides WJ11 were deleted, and the consequences in growth capacity, lipid accumulation, and metabolism were analyzed. The results showed that deletion of sodit-a or/and sodit-b reduced the extracellular malate, confirming that the products of both genes participate in malate transportation. In parallel, the lipid contents in mutants increased approximately 10-40% higher than that in the control strain, suggesting that the defect in plasma membrane malate transport results in an increase of malate available for lipid biosynthesis. Furthermore, transcriptional analysis showed that the expression levels of multiple key genes involved in the lipid biosynthesis were also increased in the knockout mutants. To the best of our knowledge, this is the first report that demonstrated the association between plasma membrane malate transporters and lipid accumulation in M. circinelloides.