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Modular metabolic engineering of lysine supply for enhanced production of bacitracin in Bacillus licheniformis

  • Biotechnological products and process engineering
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Abstract

Bacitracin is a kind of macrocyclic dodecapeptide that produced by Bacillus, precursor supply served as a critical role in bacitracin production, here, the aim of this study wants to improve bacitracin production by enhancing Lysine (Lys) supply via metabolic engineering of B. licheniformis DW2, an industrial strain for bacitracin production. Firstly, exogenous addition of Lys was proven to be favorable for bacitracin production, and the strain LYS2 was attained through strengthening Lys synthetic pathways via overexpressing diaminopimelate decarboxylase LysA from B. licheniformis and diaminopimelate dehydrogenase DdH from Corynebacterium glutamicum, and the bacitracin produced by LYS2 was increased to 838.53 U/mL by 10.85%, compared with that of DW2 (756.45 U/mL). Secondly, oxaloacetate, the precursor of Lys, was accumulated by overexpressing pyruvate carboxylase PycA in LYS2, and 17.06% increase of bacitracin yield was attained in LYS3 (885.53 U/mL), compared with DW2. Thirdly, lysine decarboxylase gene yaaO was deleted to weaken Lys degradation, and the attained strain LYS4 showed further increased bacitracin production from 885.53 to 923.43 U/mL. Lastly, the transporter LysE was confirmed to act as a Lys exporter; LysP and YvsH were identified as the Lys importers in B. licheniformis DW2, and bacitracin yield was increased to 975.43 U/mL by 28.95% in final strain LYS5 via engineering the Lys transporters. Taken together, this study implied that metabolic engineering of Lys supply modules is an efficient strategy for enhancement production of bacitracin, and provided a promising strain of B. licheniformis for industrial production of bacitracin.

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Funding

This work was supported by the National Key Research and Development Program of China (2018YFA0900303, 2015CB150505), the Technical Innovation Special Fund of Hubei Province (2018ACA149), and China Postdoctoral Science Foundation (2018M642802).

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Authors and Affiliations

  1. State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, 368 Youyi Avenue, Wuchang District, Wuhan, 430062, Hubei, People’s Republic of China

    Fei Wu, Dongbo Cai, Lingfeng Li, Yang Li, Xin Ma & Shouwen Chen

  2. Chengdu Slan Biotechnology Co. Ltd, Chengdu Hi Tech Development Zone, Chengdu, 610041, People’s Republic of China

    Hanbo Yang

  3. Lifecome Biochemistry Co. Ltd, Nanping, 353400, People’s Republic of China

    Junhui Li

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  1. Fei Wu
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  2. Dongbo Cai
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  4. Yang Li
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  5. Hanbo Yang
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  6. Junhui Li
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  7. Xin Ma
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  8. Shouwen Chen
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Contributions

D Cai and S Chen designed the study. Wu F, D Cai, L Li, and Y Li carried out the molecular biology studies and construction of engineering strains. F Wu, L Li, and Y Li carried out the fermentation studies. F, Wu, D Cai, H Yang, J Li, X Ma, and S Chen analyzed the data and wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Dongbo Cai or Shouwen Chen.

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Wu, F., Cai, D., Li, L. et al. Modular metabolic engineering of lysine supply for enhanced production of bacitracin in Bacillus licheniformis. Appl Microbiol Biotechnol 103, 8799–8812 (2019). https://doi.org/10.1007/s00253-019-10110-y

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