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Engineering a Novel Metabolic Pathway for Improving Cellular Malonyl-CoA Levels in Escherichia coli

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Abstract

Cellular pool of malonyl-CoA in Escherichia coli is small, which impedes its utility for overproduction of natural products such as phenylpropanoids, polyketides, and flavonoids. In this study, we report the use of a new metabolic pathway to increase the malonyl-CoA concentration as a limiting metabolite in E. coli. For this purpose, the malonate/sodium symporter from Malonomonas rubra, and malonyl-CoA synthetase (MCS) from Bradyrhizobium japonicum were co-expressed in E. coli. This new pathway allows the cell to actively import malonate from the culture medium and to convert malonate and CoA to malonyl-CoA via an ATP-dependent ligation reaction. HPLC analysis confirmed elevated levels of malonyl-CoA and (2S)-naringenin as a malonyl-CoA-dependent metabolite, in E. coli. A 6.8-fold and more than 3.5-fold increase in (2S)-naringenin production were achieved in the engineered host in comparison with non-engineered E. coli and previously reported passive transport MatBMatC pathway, respectively. This observation suggests that using active transporters of malonate not only improves malonyl-CoA-dependent production but also makes it possible to harness low concentrations of malonate in culture media.

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Data availability

The published article includes all data sets generated during this study.

Abbreviations

CoA:

Coenzyme A

MCoA:

Malonyl-coenzyme A

MCS:

Malonyl-CoA synthetase

ACC:

Acetyl-CoA carboxylase

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Acknowledgements

We gratefully appreciate the Iran National Science Foundation (INSF) for financial support of this research (Project No. 94017574). Additionally, we would like to thank Prof. K.M. Pos (Goethe University Frankfurt, Germany) and Prof. J. Chen (Jiangnan University, Jiangsu, China) for providing us with the recombinant vectors and Mohammad Ranjbar for his constructive feedbacks over the manuscript.

Funding

This study was supported by Iran National Science Foundation (Grant No. 94017574).

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Authors and Affiliations

  1. Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran

    Mohammad Hossein Moteallehi-Ardakani, Sedigheh Asad, Sayed-Amir Marashi, Afrooz Moghaddasi & Parisa Zarparvar

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  1. Mohammad Hossein Moteallehi-Ardakani
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  2. Sedigheh Asad
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Moteallehi-Ardakani, M.H., Asad, S., Marashi, SA. et al. Engineering a Novel Metabolic Pathway for Improving Cellular Malonyl-CoA Levels in Escherichia coli. Mol Biotechnol 65, 1508–1517 (2023). https://doi.org/10.1007/s12033-022-00635-5

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