Department of Biological Science, Vanderbilt University, 465 21st Ave S, Nashville, TN, 37240, USA.
Department of Biology and Chemistry, Texas A&M International University, 5201 University Blvd, Laredo, TX, 78041, USA.
BMC Biotechnol. 2023 Sep 8;23(1):35. doi: 10.1186/s12896-023-00808-3.
Biofuel research that aims to optimize growth conditions in microalgae is critically important. Chlamydomonas reinhardtii is a green microalga that offers advantages for biofuel production research. This study compares the effects of nitrogen-, sulfur-, and nitrogen and sulfur- deprivations on the C. reinhardtii starchless mutant cc5373-sta6. Specifically, it compares growth, lipid body accumulation, and expression levels of acetyl-CoA carboxylase (ACC) and phosphoenolpyruvate carboxylase (PEPC).
Among nutrient-deprived cells, TAP-S cells showed significantly higher total chlorophyll, cell density, and protein content at day 6 (p < 0.05). Confocal analysis showed a significantly higher number of lipid bodies in cells subjected to nutrient deprivation than in the control over the course of six days; N deprivation for six days significantly increased the size of lipid bodies (p < 0.01). In comparison with the control, significantly higher ACC expression was observed after 8 and 24 h of NS deprivation and only after 24 h with N deprivation. On the other hand, ACC and PEPC expression at 8 and 24 h of S deprivation was not significantly different from that in the control. A significantly lower PEPC expression was observed after 8 h of N and NS deprivation (p < 0.01), but a significantly higher PEPC expression was observed after 24 h (p < 0.01).
Based on our findings, it would be optimum to cultivate cc5373-sta6 cells in nutrient deprived conditions (-N, -S or -NS) for four days; whereby there is cell growth, and both a high number of lipid bodies and a larger size of lipid bodies produced.
旨在优化微藻生长条件的生物燃料研究至关重要。莱茵衣藻是一种绿色微藻,可为生物燃料生产研究提供优势。本研究比较了氮、硫和氮硫剥夺对莱茵衣藻无淀粉突变体 cc5373-sta6 的影响。具体来说,比较了生长、脂体积累以及乙酰辅酶 A 羧化酶(ACC)和磷酸烯醇丙酮酸羧化酶(PEPC)的表达水平。
在营养剥夺细胞中,TAP-S 细胞在第 6 天显示出明显更高的总叶绿素、细胞密度和蛋白质含量(p<0.05)。共聚焦分析显示,在六天的过程中,与对照相比,营养剥夺细胞中的脂体数量明显更多;六天的氮剥夺显著增加了脂体的大小(p<0.01)。与对照相比,在 NS 剥夺 8 和 24 小时后观察到 ACC 表达显著升高,而仅在氮剥夺 24 小时后观察到 ACC 表达升高。另一方面,在 S 剥夺 8 和 24 小时时,ACC 和 PEPC 的表达与对照无显著差异。氮和 NS 剥夺 8 小时后观察到 PEPC 表达显著降低(p<0.01),但 24 小时后观察到 PEPC 表达显著升高(p<0.01)。
根据我们的发现,在营养剥夺条件(-N、-S 或 -NS)下培养 cc5373-sta6 细胞四天是最佳的;在此期间会发生细胞生长,并且产生大量的脂体和更大的脂体。