Quantitative analysis of an engineered CO 2-fixing Escherichia coli reveals great potential of heterotrophic CO 2 fixation

Biotechnology for Biofuels

Fuyu Gong¹  Guoxia Liu²  Xiaoyun Zhai¹  Jie Zhou²  Zhen Cai²  Yin Li² 

[1] University of the Chinese Academy of Sciences, Beijing, China

[2] CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road Chaoyang District, Beijing 100101, China

关键词: Rubisco;    Carbonic anhydrase;    Heterotrophic microbe;    CO2-fixation rate;    Carbon fixation;   

Others  :  1219216
DOI  :  10.1186/s13068-015-0268-1

 received in 2015-02-23, accepted in 2015-06-05,  发布年份 2015

【 摘 要 】

Background

Production of fuels from the abundant and wasteful CO ₂is a promising approach to reduce carbon emission and consumption of fossil fuels. Autotrophic microbes naturally assimilate CO ₂using energy from light, hydrogen, and/or sulfur. However, their slow growth rates call for investigation of the possibility of heterotrophic CO ₂fixation. Although preliminary research has suggested that CO ₂fixation in heterotrophic microbes is feasible after incorporation of a CO ₂ -fixing bypass into the central carbon metabolic pathway, it remains unclear how much and how efficient that CO ₂can be fixed by a heterotrophic microbe.

Results

A simple metabolic flux index was developed to indicate the relative strength of the CO ₂ -fixation flux. When two sequential enzymes of the cyanobacterial Calvin cycle were incorporated into an E. coli strain, the flux of the CO ₂ -fixing bypass pathway accounts for 13 % of that of the central carbon metabolic pathway. The value was increased to 17 % when the carbonic anhydrase involved in the cyanobacterial carbon concentrating mechanism was introduced, indicating that low intracellular CO ₂concentration is one limiting factor for CO ₂fixation in E. coli. The engineered CO ₂ -fixing E. coli with carbonic anhydrase was able to fix CO ₂at a rate of 19.6 mg CO ₂L ⁻¹  h ⁻¹or the specific rate of 22.5 mg CO ₂g DCW ⁻¹  h ⁻¹ . This CO ₂ -fixation rate is comparable with the reported rates of 14 autotrophic cyanobacteria and algae (10.5–147.0 mg CO ₂L ⁻¹  h ⁻¹or the specific rates of 3.5–23.7 mg CO ₂g DCW ⁻¹  h ⁻¹ ).

Conclusions

The ability of CO ₂fixation was created and improved in E. coli by incorporating partial cyanobacterial Calvin cycle and carbon concentrating mechanism, respectively. Quantitative analysis revealed that the CO ₂ -fixation rate of this strain is comparable with that of the autotrophic cyanobacteria and algae, demonstrating great potential of heterotrophic CO ₂fixation.

【 授权许可】

   
2015 Gong et al.

【 图 表 】

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