Avidan Omri, Brandis Alexander, Rogachev Ilana, Pick Uri
Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel
Biological Services Unit, The Weizmann Institute of Science, Rehovot 76100, Israel.
J Exp Bot. 2015 Jul;66(13):3725-35. doi: 10.1093/jxb/erv166. Epub 2015 Apr 28.
Triglycerides (TAGs) from microalgae can be utilized as food supplements and for biodiesel production, but little is known about the regulation of their biosynthesis. This work aimed to test the relationship between acetyl-CoA (Ac-CoA) levels and TAG biosynthesis in green algae under nitrogen deprivation. A novel, highly sensitive liquid chromatography mass spectrometry (LC-MS/MS) technique enabled us to determine the levels of Ac-CoA, malonyl-CoA, and unacetylated (free) CoA in green microalgae. A comparative study of three algal species that differ in TAG accumulation levels shows that during N starvation, Ac-CoA levels rapidly rise, preceding TAG accumulation in all tested species. The levels of Ac-CoA in the high TAG accumulator Chlorella desiccata exceed the levels in the moderate TAG accumulators Dunaliella tertiolecta and Chlamydomonas reinhardtii. Similarly, malonyl-CoA and free CoA levels also increase, but to lower extents. Calculated cellular concentrations of Ac-CoA are far lower than reported K mAc-CoA values of plastidic Ac-CoA carboxylase (ptACCase) in plants. Transcript level analysis of plastidic pyruvate dehydrogenase (ptPDH), the major chloroplastic Ac-CoA producer, revealed rapid induction in parallel with Ac-CoA accumulation in C. desiccata, but not in D. tertiolecta or C. reinhardtii. It is proposed that the capacity to accumulate high TAG levels in green algae critically depends on their ability to divert carbon flow towards Ac-CoA. This requires elevation of the chloroplastic CoA pool level and enhancement of Ac-CoA biosynthesis. These conclusions may have important implications for future genetic manipulation to enhance TAG biosynthesis in green algae.
微藻中的甘油三酯(TAGs)可作为食品补充剂并用于生物柴油生产,但人们对其生物合成的调控知之甚少。这项工作旨在测试氮缺乏条件下绿藻中乙酰辅酶A(Ac-CoA)水平与TAG生物合成之间的关系。一种新颖、高灵敏度的液相色谱质谱联用(LC-MS/MS)技术使我们能够测定绿微藻中Ac-CoA、丙二酰辅酶A和未乙酰化(游离)辅酶A的水平。对三种TAG积累水平不同的藻类进行的比较研究表明,在氮饥饿期间,Ac-CoA水平迅速上升,先于所有测试物种中的TAG积累。高TAG积累者小球藻(Chlorella desiccata)中的Ac-CoA水平超过了中等TAG积累者盐生杜氏藻(Dunaliella tertiolecta)和莱茵衣藻(Chlamydomonas reinhardtii)中的水平。同样,丙二酰辅酶A和游离辅酶A水平也会增加,但增幅较小。计算得出的细胞内Ac-CoA浓度远低于报道的植物中质体Ac-CoA羧化酶(ptACCase)的KmAc-CoA值。对主要的叶绿体Ac-CoA生产者质体丙酮酸脱氢酶(ptPDH)的转录水平分析表明,在小球藻中,ptPDH与Ac-CoA积累同时迅速诱导,但在盐生杜氏藻或莱茵衣藻中则不然。有人提出,绿藻积累高TAG水平的能力关键取决于它们将碳流转向Ac-CoA 的能力。这需要提高叶绿体辅酶A库水平并增强Ac-CoA生物合成。这些结论可能对未来通过基因操作提高绿藻中TAG生物合成具有重要意义。