Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York 11794.
Biology Department, Brookhaven National Laboratory, Upton, New York 11973.
Plant Physiol. 2019 Jul;180(3):1351-1361. doi: 10.1104/pp.19.00396. Epub 2019 May 13.
Cyclopropane fatty acids (CPAs) are useful feedstocks for biofuels and bioproducts such as lubricants and biodiesel. Our goal is to identify factors that can facilitate the accumulation of CPA in seed triacylglycerol (TAG) storage oil. We hypothesized that the poor metabolism of CPA through the TAG biosynthetic network could be overcome by the addition of enzymes from species that naturally accumulate CPA in their seed oil, such as lychee (), which contains approximately 40% CPA in TAG. Our previous work on engineering CPA accumulation in crop and model plants identified a metabolic bottleneck between phosphatidylcholine (PC), the site of CPA biosynthesis, diacylglycerol (DAG), and TAG. Here, we report the cloning and heterologous expression in camelina () of a lychee (), which encodes the enzyme that catalyzes the transfer of the phosphocholine headgroup from PC to DAG. Camelina lines coexpressing and () showed up to a 50% increase of CPA in mature seed, relative to the background. Stereospecific lipid compositional analysis showed that the expression of strongly reduced the level of C18:1 substrate at PC-1 and PC--2 (i.e. the sites of CPA synthesis), while the levels of CPA increased in PC-2, DAG-1 and DAG--2, and both -1/3 and -2 positions in TAG. Taken together, these data suggest that the addition of PDCT facilitates more efficient movement of CPA from PC to DAG and establishes LcPDCT as a useful factor to combine with others to enhance CPA accumulation in plant seed oil.
环丙烷脂肪酸 (CPAs) 是生物燃料和生物制品(如润滑剂和生物柴油)的有用原料。我们的目标是确定可以促进 CPAs 在种子三酰基甘油 (TAG) 储存油中积累的因素。我们假设,通过 TAG 生物合成途径对 CPAs 的代谢不良可以通过添加来自天然在其种子油中积累 CPAs 的物种的酶来克服,例如荔枝 (),其 TAG 中含有约 40%的 CPAs。我们之前在工程作物和模式植物中积累 CPA 的工作确定了一个代谢瓶颈,位于磷脂酰胆碱 (PC),即 CPA 生物合成的部位,二酰基甘油 (DAG) 和 TAG 之间。在这里,我们报告了在荠蓝 ()中克隆和异源表达荔枝 () 的,该基因编码催化从 PC 向 DAG 转移磷酸胆碱头基的酶。共表达 和 ()的荠蓝系中,CPA 在成熟种子中的含量比 背景增加了 50%。立体特异性脂质成分分析表明, 的表达强烈降低了 PC-1 和 PC--2 处的 C18:1 底物水平(即 CPA 合成部位),而 PC-2、DAG-1 和 DAG--2 以及 -1/3 和 -2 位置的 CPA 水平增加。总的来说,这些数据表明添加 PDCT 有助于更有效地将 CPA 从 PC 转移到 DAG,并将 LcPDCT 确立为一种有用的因子,可与其他因子结合使用,以提高植物种子油中的 CPA 积累。