Construction of pyruvate producing strain with intact pyruvate dehydrogenase and genome-wide transcription analysis

Original Paper

Abstract

To obtain strain YP211 with a high tendency for accumulating pyruvate, central metabolic pathways were modified in Escherichia coli MG1655. Specifically, seven genes (ldhA, pflB, pta-ackA, poxB, ppc, frdBC) were knocked out sequentially and full pyruvate dehydrogenase was retained. In batch fermentation with M9 medium, pyruvate yield and production rate reached 0.63 g/g glucose and 1.89 g/(1 h), respectively. Meanwhile, the production of acetate, succinate, and other carboxylates was effectively controlled. To understand the physiological observations, we further completed genome-wide transcription analysis of wild-type and YP211. As the acetic acid pathways were blocked, the pathways of convertion of pyruvate to phosphoenol pyruvate and acetyl CoA were enhanced. The transcription of pck, as an alternative gene for ppc, was increased by 2.6 times. So even if gene ppc was inactivated, the tricarboxylic acid pathway was still enhanced in YP211. In order to balance intracellular NADH/NAD+, oxidative phosphorylation and flagellar assembly system were also up-regulated significantly.

Graphical Abstract

Biochemical pathways involved in pyruvate accumulation in YP211 (a). Transcriptional differences of genes related to pyruvate metabolism between strain YP211 and E. coli wild-type (b).

Keywords

Pyruvate Escherichia coli Transcription analysis Oxidation–Reduction Flagellar assembly 

Notes

Acknowledgements

The work was supported by the National High Technology Research and Development Program of China, No. 2014AA021900, and the Foundation (No. 2015IM002) of Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education and Tianjin Key Lab of Industrial Microbiology (Tianjin University of Science & Technology).

Supplementary material

11274_2016_2202_MOESM1_ESM.docx (47 kb)
Supplementary material 1 (DOCX 46 KB)

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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.Institute of Process EngineeringChinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.Institute of Agro-food Science and TechnologyShandong Academy of Agricultural SciencesJi’nanPeople’s Republic of China

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