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Distinct Promoters Affect Pyrroloquinoline Quinone Production in Recombinant Escherichia coli and Klebsiella pneumoniae

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Abstract

Pyrroloquinoline quinone (PQQ) is a versatile quinone cofactor participating in numerous biological processes. Klebsiella pneumoniae can naturally synthesize PQQ for harboring intact PQQ synthesis genes. Previous metabolic engineering of K. pneumoniae failed to overproduce PQQ due to the employment of strong promoter in expression vector. Here we report that a moderate rather than strong promoter is efficient for PQQ production. To screen an appropriate promoter, a total of four distinct promoters—lac promoter, pk promoter of glycerol dehydratase gene (dhaB1), promoter of kanamycin resistance gene, and T7 promoter (as the control)—were individually used for overexpressing the endogenous PQQ genes in K. pneumoniae along with heterologous expression in Escherichia coli. We found that all recombinant K. pneumoniae strains produced more PQQ than recombinant E. coli strains that carried corresponding vectors, indicating that K. pneumoniae is superior to E. coli for the production of PQQ. Particularly, the recombinant K. pneumoniae recruiting the promoter of kanamycin resistance gene produced the highest PQQ (1,700 nmol), revealing that a moderate rather than strong promoter is efficient for PQQ production. Furthermore, PQQ production was roughly proportional to glucose concentration increasing from 0.5 to 1.5 g/L, implying the synergism between PQQ biosynthesis and glucose utilization. This study not only provides a feasible strategy for production of PQQ in K. pneumoniae, but also reveals the exquisite synchronization among PQQ biosynthesis, glucose metabolism, and cell proliferation.

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Acknowledgments

This work was supported by National Natural Science Foundation of China (No. 21076013, 21276014) and National Basic Research Program of China (973 Program) (2012CB725200).

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Authors and Affiliations

  1. Beijing Key Lab of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, People’s Republic of China

    Jiguo Sun, Zengye Han & Pingfang Tian

  2. College of Biochemical Engineering, Beijing Union University, Beijing, 100023, People’s Republic of China

    Xizhen Ge

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  1. Jiguo Sun
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  2. Zengye Han
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  3. Xizhen Ge
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  4. Pingfang Tian
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Correspondence to Pingfang Tian.

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Jiguo Sun and Zengye Han contributed equally to this work.

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Sun, J., Han, Z., Ge, X. et al. Distinct Promoters Affect Pyrroloquinoline Quinone Production in Recombinant Escherichia coli and Klebsiella pneumoniae . Curr Microbiol 69, 451–456 (2014). https://doi.org/10.1007/s00284-014-0607-7

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  • DOI: https://doi.org/10.1007/s00284-014-0607-7

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