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Engineering the growth pattern and cell morphology for enhanced PHB production by Escherichia coli

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Abstract

E. coli JM109∆envC∆nlpD deleted with genes envC and nlpD responsible for degrading peptidoglycan (PG) led to long filamentous cell shapes. When cell fission ring location genes minC and minD of Escherichia coli were deleted, E. coli JM109∆minCD changed the cell growth pattern from binary division to multiple fissions. Bacterial morphology can be further engineered by overexpressing sulA gene resulting in inhibition on FtsZ, thus generating very long cellular filaments. By overexpressing sulA in E. coli JM109∆envC∆nlpD and E. coli JM109∆minCD harboring poly(3-hydroxybutyrate) (PHB) synthesis operon phbCAB encoded in plasmid pBHR68, respectively, both engineered cells became long filaments and accumulated more PHB compared with the wild-type. Under same shake flask growth conditions, E. coli JM109∆minCD (pBHR68) overexpressing sulA grown in multiple fission pattern accumulated approximately 70 % PHB in 9 g/L cell dry mass (CDM), which was significantly higher than E. coli JM109∆envC∆nlpD and the wild type, that produced 7.6 g/L and 8 g/L CDM containing 64 % and 51 % PHB, respectively. Results demonstrated that a combination of the new division pattern with elongated shape of E. coli improved PHB production. This provided a new vision on the enhanced production of inclusion bodies.

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Acknowledgments

We are grateful to the Center of Biomedical Analysis, Tsinghua University for the SEM and TEM studies. Plasmid pBHR68 was kindly donated by Professor Alexander Steinbűchel of Műnster University in Germany. This research was financially supported by the State Basic Science Foundation 973 (Grant no. 2012CB725201) and National Natural Science Foundation of China (Grant no. 31430003 and 31270146).

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Authors and Affiliations

  1. Center for Synthetic and Systems Biology, School of Life Science, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, 100084, China

    Hong Wu, Jinchun Chen & Guo-Qiang Chen

  2. Center for Nano and Micro Mechanics, Tsinghua University, Beijing, 100084, China

    Guo-Qiang Chen

  3. MOE Key Lab of Industrial Biocatalysis, Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China

    Guo-Qiang Chen

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  1. Hong Wu
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Correspondence to Guo-Qiang Chen.

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Wu, H., Chen, J. & Chen, GQ. Engineering the growth pattern and cell morphology for enhanced PHB production by Escherichia coli . Appl Microbiol Biotechnol 100, 9907–9916 (2016). https://doi.org/10.1007/s00253-016-7715-1

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