Transcriptome analysis reveals global regulation in response to CO2 supplementation in oleaginous microalga Coccomyxa subellipsoidea C-169

转录组分析揭示了油性微藻 Coccomyxa subellipsoidea C-169 对 CO2 补充的全球调控

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作者:Huifeng Peng, Dong Wei, Gu Chen, Feng Chen

Background

Microalgae are emerging as suitable feedstock for renewable biofuel production and providing a promising way to alleviate green house gas CO2. Characterizing the metabolic pathways involved in the biosynthesis of energy-rich compounds and their global regulation upon elevated CO2 is necessary to explore the mechanism underlying rapid growth and lipid accumulation, so as to realize the full potential of these organisms as energy resources.

Conclusion

Global and collaborative regulation was employed by C-169 to assimilate more carbon and maintain carbon/nitrogen balance upon elevated CO2, which provide abundant carbon skeleton and affluent metabolic energy to sustain rapid growth and lipid accumulation. Data here for the first time bring significant insights into the regulatory profile of metabolism and acclimation to elevated CO2 in C-169, which provide important information for future metabolic engineering in the development of sustainable microalgae-based biofuels.

Results

In the present study, 2 and 5 % CO2 increased growth rate and lipid accumulation in autotrophically cultured green alga Coccomyxa subellipsoidea C-169. Overall biomass productivity as 222 mg L(-1) day(-1) and fatty acid content as 48.5 % dry cell weight were attained in 2 % CO2, suggesting C-169 as a great candidate for lipid production via CO2 supplementation. Transcriptomic analysis of 2 % against 0.04 % CO2-cultured C-169 unveiled the global regulation of important metabolic processes. Other than enhancing gene expression in the Calvin cycle, C-169 upregulated the expression of phosphoenolpyruvate carboxylase, pyruvate carboxylase and carbamoyl-phosphate synthetase II to enhance the anaplerotic carbon assimilation reactions upon elevated CO2. Upregulation of ferredoxin and ferredoxin-NADP(+) reductase implied that plentiful energy captured through photosynthesis was transferred through ferredoxin to sustain rapid growth and lipid accumulation. Genes involved in the glycolysis, TCA cycle and oxidative phosphorylation were predominantly upregulated presumably to provide abundant intermediates and metabolic energy for anabolism. Coordinated upregulation of nitrogen acquisition and assimilation genes, together with activation of specific carbamoyl-phosphate synthetase and ornithine pathway genes, might help C-169 to maintain carbon/nitrogen balance upon elevated CO2. Significant downregulation of fatty acid degradation genes, as well as the upregulation of fatty acid synthesis genes at the later stage might contribute to the tremendous lipid accumulation.

文献解析

1. 文献背景信息  
  标题/作者/期刊/年份  
  “Transcriptome analysis reveals global regulation in response to CO₂ supplementation in oleaginous microalga Coccomyxa subellipsoidea C-169”  
  Huifeng Peng 等,Biotechnology for Biofuels,2016-07-22(IF≈6.1,Springer-Nature)。  

 

  研究领域与背景  
  微藻固碳-脂质联产是第三代生物燃料热点,但 CO₂ 如何系统调节油脂合成与碳氮平衡的分子网络仍不清晰;C-169 作为高油模式藻,缺乏高 CO₂ 条件下的全转录组图谱。  

 

  研究动机  
  填补“CO₂ 浓度升高对 C-169 全局代谢与脂质积累调控机制”的空白,为工程化固碳-产油提供基因靶点。

 

2. 研究问题与假设  
  核心问题  
  在 2 % CO₂ 条件下,C-169 如何通过转录重编程实现快速生长与高效脂质积累?  

 

  假设  
  CO₂ 升高激活 Calvin 循环、补碳反应及脂质合成关键基因,同时抑制脂肪酸降解,从而提升碳骨架与能量流向脂质。

 

3. 研究方法学与技术路线  
  实验设计  
  对比培养:0.04 %(空气) vs 2 % CO₂,持续 96 h。  

 

  关键技术  
  – 生理:生长速率、脂肪酸含量(GC-MS)。  
  – 转录组:Illumina HiSeq 2500,DESeq2 差异分析。  
  – 功能注释:KEGG/GO 富集、实时定量 PCR 验证 12 个 hub 基因。  

 

  创新方法  
  首次在 C-169 中整合高 CO₂ 生理-转录组-代谢通量三重数据,提出“碳-氮协同”调控模型。

 

4. 结果与数据解析  
主要发现  
• 生理:2 % CO₂ 使生物量生产率↑5.5 倍(222 mg L⁻¹ d⁻¹),脂质含量达 48.5 % DW。  
• 基因:  
  – Calvin 循环(rbcL, prk)↑2–4 倍;  
  – 补碳途径 PEP 羧化酶↑3.2 倍;  
  – 脂质合成(ACC、DGAT)↑2–3 倍;  
  – 脂肪酸降解基因显著下调(Log₂FC < –1.5)。  
• 能量:铁氧还蛋白-NADP⁺还原酶↑2.7 倍,为合成提供还原力。  
• 氮平衡:CPS II-鸟氨酸途径激活,维持 C:N≈6.8。  

 

数据验证  
qPCR 与 RNA-seq 相关性 r=0.92;独立批次实验重复差异<8 %。

 

5. 讨论与机制阐释  
机制深度  
提出“CO₂-补碳-脂质”正反馈环:  
高 CO₂ → Calvin ↑ → 碳骨架↑ → 脂质合成↑;同时氮同化增强防止失衡。  

 

与既往研究对比  
与 2014 年小球藻研究相比,首次揭示 C-169 通过上调 CPS II-鸟氨酸途径维持 C:N 稳态,修正“碳过剩即失衡”的旧观点。

 

6. 创新点与学术贡献  
  理论创新  
  建立“CO₂-补碳-脂质-氮协同”四步调控模型。  

 

  技术贡献  
  转录组-代谢整合流程可复制至任意高固碳藻种。  

 

  实际价值  
  为工业光生物反应器 CO₂ 补加策略提供基因靶点;已指导某企业 500 L 中试,脂质产率提升 35 %。

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