State Key Laboratory of Urban Water Resource and Environment, School of Environment, Harbin Institute of Technology, P.O. Box 2614, 73 Huanghe Road, Harbin 150090, China.
J Hazard Mater. 2019 Aug 5;375:115-120. doi: 10.1016/j.jhazmat.2019.04.079. Epub 2019 Apr 24.
Currently, the complex mechanism of lipid production in microalgal cells is still unclear, and the platform suitable for microalgal genetic transformation is urgent to be established. In this study, the whole genome of a lipid-rich microalgal mutant strain Scenedesmus sp. Z-4 and a lipid-poor wild strain Scenedesmus sp. MC-1 were sequenced, and results revealed that the sequences of 1,256 genes were changed and 148 differential genes related to glucose and lipid metabolism were identified. Especially, gene differentiation of acetyl-CoA carboxylase (ACCase) and phosphoenolpyruvate carboxylase (PEPC) in mutant strain Z-4 and wild strain MC-1, which played key roles in lipid synthesis, were evaluated. Furthermore, to investigate whether mutated ACCase and PEPC genes affect the lipid production, two genes from mutant strain Z-4 were transformed into the expression system of wild strain MC-1. Nine transformants with higher lipid content were successfully obtained, in which the optimal transformant with 28.6% more intracellular lipid than wild strain MC-1 was isolated by overexpression of mutated ACCase gene, demonstrating the important role of ACCase in lipid accumulation of microalgal cells. These results could provide a better understanding of the superior lipid production of mutant strain Scenedesmus sp. Z-4.
目前,微藻细胞中脂质产生的复杂机制尚不清楚,适合微藻遗传转化的平台也亟待建立。本研究对富脂微藻突变株 Scenedesmus sp. Z-4 和贫脂野生株 Scenedesmus sp. MC-1 的全基因组进行了测序,结果表明有 1,256 个基因序列发生了变化,并鉴定出了 148 个与葡萄糖和脂质代谢相关的差异基因。特别是,突变株 Z-4 和野生株 MC-1 中乙酰辅酶 A 羧化酶 (ACCase) 和磷酸烯醇式丙酮酸羧化酶 (PEPC) 的基因分化,这两个基因在脂质合成中起着关键作用,对其进行了评估。此外,为了研究突变的 ACCase 和 PEPC 基因是否影响脂质的产生,将突变株 Z-4 中的两个基因转入野生株 MC-1 的表达系统中。成功获得了 9 株具有较高脂质含量的转化株,其中通过过表达突变的 ACCase 基因,获得了一株比野生株 MC-1 细胞内脂质含量高 28.6%的最优转化株,证明了 ACCase 在微藻细胞脂质积累中的重要作用。这些结果可以更好地理解突变株 Scenedesmus sp. Z-4 具有优越的脂质生产能力。