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Pyruvate decarboxylase and alcohol dehydrogenase overexpression in Escherichia coli resulted in high ethanol production and rewired metabolic enzyme networks

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Abstract

Pyruvate decarboxylase and alcohol dehydrogenase are efficient enzymes for ethanol production in Zymomonas mobilis. These two enzymes were over-expressed in Escherichia coli, a promising candidate for industrial ethanol production, resulting in high ethanol production in the engineered E. coli. To investigate the intracellular changes to the enzyme overexpression for homoethanol production, 2-DE and LC–MS/MS were performed. More than 1,000 protein spots were reproducibly detected in the gel by image analysis. Compared to the wild-type, 99 protein spots showed significant changes in abundance in the recombinant E. coli, in which 46 were down-regulated and 53 were up-regulated. Most proteins related to tricarboxylic acid cycle, glycerol metabolism and other energy metabolism were up-regulated, whereas proteins involved in glycolysis and glyoxylate pathway were down-regulated, indicating the rewired metabolism in the engineered E. coli. As glycolysis is the main pathway for ethanol production, and it was inhibited significantly in engineered E. coli, further efforts should be directed at minimizing the repression of glycolysis to optimize metabolism network for higher yields of ethanol production.

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Acknowledgments

This work was supported by grants from Beijing Natural Science Foundation (Nos. 5132004, 6122005), National Natural Science Foundation of China (Nos. 30970250, 31370674), China Postdoctoral Science Foundation to Dr. Wang, and Science and Technology Platform Construction Program of Beijing Municipal Commission of Education (PXM2012_014207_000028) and Beijing University of Agriculture (2014).

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Authors and Affiliations

  1. Key Laboratory of Urban Agriculture (North) Ministry of Agriculture, Beijing University of Agriculture, Beijing, 102206, China

    Mingfeng Yang, Chunya Bu, Guanglu Shi & Xiaoqin Wang

  2. College of Life Sciences, Capital Normal University, Beijing, 100048, China

    Xuefeng Li, Hui Wang, Xiushan Yang, Yong Hu & Xiaoqin Wang

Authors
  1. Mingfeng Yang
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  2. Xuefeng Li
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  3. Chunya Bu
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  4. Hui Wang
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  5. Guanglu Shi
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  6. Xiushan Yang
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  7. Yong Hu
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  8. Xiaoqin Wang
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Corresponding authors

Correspondence to Yong Hu or Xiaoqin Wang.

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Mingfeng Yang and Xuefeng Li have contributed equally to this work.

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Yang, M., Li, X., Bu, C. et al. Pyruvate decarboxylase and alcohol dehydrogenase overexpression in Escherichia coli resulted in high ethanol production and rewired metabolic enzyme networks. World J Microbiol Biotechnol 30, 2871–2883 (2014). https://doi.org/10.1007/s11274-014-1713-1

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  • DOI: https://doi.org/10.1007/s11274-014-1713-1

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