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Metabolic engineering of Escherichia coli for secretory production of free haem

Nature Catalysis volume 1, pages 720–728 (2018)Cite this article

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Abstract

Haem has widespread applications in healthcare and food supplement industries. Escherichia coli has previously been engineered to produce a small amount of haem intracellularly through the C4 pathway, requiring extraction for applications. Here we report secretory production of free haem by engineered E. coli strains, using the C5 pathway and the optimized downstream pathway for haem biosynthesis. Furthermore, knocking out ldhA, pta and also yfeX—encoding a putative haem-degrading enzyme—results in 7.88 mg l−1 of total haem with 1.26 mg l−1 of extracellular haem in flask cultivation. Fed-batch fermentations of the engineered strain overexpressing a haem exporter CcmABC from glucose only and glucose supplemented with l-glutamate secrete 73.4 and 151.4 mg l−1 of haem, respectively, which are 63.5% of 115.5 mg l−1 and 63.3% of 239.2 mg l−1 of total haem produced. The engineered E. coli strain reported here will be useful for microbial production of free haem.

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Fig. 1: Haem biosynthetic pathways examined for secretory production of free haem.
Fig. 2: The scheme of engineering strategies to optimize haem production in E. coli.
Fig. 3: Flask cultivation results of the recombinant E. coli strains producing haem.
Fig. 4: Averaged batch fermentation profile of the strain HAEM6.
Fig. 5: Averaged fed-batch fermentation profiles of the strain HAEM6.
Fig. 6: Averaged fed-batch fermentation profiles of the strain HAEM7.

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Acknowledgements

We thank Won Jun Kim for discussion. This work was supported by the Technology Development Program to Solve Climate Changes on Systems Metabolic Engineering for Biorefineries (NRF-2012M1A2A2026556 and NRF-2012M1A2A2026557) from the Ministry of Science and ICT, through the National Research Foundation (NRF) of Korea.

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Authors and Affiliations

  1. Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea

    Xin Rui Zhao, Kyeong Rok Choi & Sang Yup Lee

  2. BioProcess Engineering Research Center, KAIST, Daejeon, Republic of Korea

    Sang Yup Lee

  3. BioInformatics Research Center, KAIST, Daejeon, Republic of Korea

    Sang Yup Lee

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  1. Xin Rui Zhao
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Contributions

X.R.Z. and S.Y.L. designed the project. X.R.Z. and K.R.C. performed the experiments and analysed the data. X.R.Z., K.R.C. and S.Y.L. wrote the manuscript together.

Corresponding author

Correspondence to Sang Yup Lee.

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Competing interests

The authors declare competing financial interests as a patent (Korean Patent Application No. 10-2017-0170185) of commercial interest entitled “Method for Producing Exracellular Haem Using Metabolically Engineered Microorganism” is filed on this technology by Korea Advanced Institute of Science and Technology. S.Y.L., X.R.Z. and K.R.C. are the inventors of this patent that covers the construction and fermentation of mircroorganisms capable of secretory production of haem reported in this manuscript.

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Zhao, X.R., Choi, K.R. & Lee, S.Y. Metabolic engineering of Escherichia coli for secretory production of free haem. Nat Catal 1, 720–728 (2018). https://doi.org/10.1038/s41929-018-0126-1

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