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Rational design of a synthetic Entner–Doudoroff pathway for enhancing glucose transformation to isobutanol in Escherichia coli

  • Metabolic Engineering and Synthetic Biology - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Isobutanol as a more desirable biofuel has attracted much attention. In our previous work, an isobutanol-producing strain Escherichia coli LA09 had been obtained by rational redox status improvement under guidance of the genome-scale metabolic model. However, the low transformation from sugar to isobutanol is a limiting factor for isobutanol production by E. coli LA09. In this study, the intracellular metabolic profiles of the isobutanol-producing E. coli LA09 with different initial glucose concentrations were investigated and the metabolic reaction of fructose 6-phosphate to 1, 6-diphosphate fructose in glycolytic pathway was identified as the rate-limiting step of glucose transformation. Thus, redesigned carbon catabolism was implemented by altering flux of sugar metabolism. Here, the heterologous Entner–Doudoroff (ED) pathway from Zymomonas mobilis was constructed, and the adaptation of upper and lower parts of ED pathway was further improved with artificial promoters to alleviate the accumulation of toxic intermediate metabolite 2-keto-3-deoxy-6-phospho-gluconate (KDPG). Finally, the best isobutanol-producing E. coli ED02 with higher glucose transformation and isobutanol production was obtained. In the fermentation of strain E. coli ED02 with 45 g/L initial glucose, the isobutanol titer, yield and average producing rate were, respectively, increased by 56.8, 47.4 and 88.1% to 13.67 g/L, 0.50 C-mol/C-mol and 0.456 g/(L × h) in a shorter time of 30 h, compared with that of the starting strain E. coli LA09.

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (No. 21676189), the key technologies R & D program of Tianjin (No. 16YFZCSY00780).

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Authors and Affiliations

  1. Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, People’s Republic of China

    Shaoxiong Liang, Hong Chen, Jiao Liu & Jianping Wen

  2. SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People’s Republic of China

    Shaoxiong Liang, Hong Chen, Jiao Liu & Jianping Wen

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  1. Shaoxiong Liang
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  2. Hong Chen
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Correspondence to Jianping Wen.

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Liang, S., Chen, H., Liu, J. et al. Rational design of a synthetic Entner–Doudoroff pathway for enhancing glucose transformation to isobutanol in Escherichia coli. J Ind Microbiol Biotechnol 45, 187–199 (2018). https://doi.org/10.1007/s10295-018-2017-5

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