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Functional characterization of a novel violacein biosynthesis operon from Janthinobacterium sp. B9-8

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Abstract

Violacein is a secondary metabolite mainly produced by Gram-negative bacteria that is formed from tryptophan by five enzymes encoded by a single operon. It is a broad-spectrum antibacterial pigment with various important biological activities such as anti-tumor, antiviral, and antioxidative effects. The newly discovered violacein operon vioABCDE was identified in the genome of the extremophile Janthinobacterium sp. B9-8. The key enzyme-encoding genes were cloned to construct the multigene coexpression plasmids pET-vioAB and pRSF-vioCDE. The violacein biosynthesis pathway was heterologously introduced into engineered Escherichia coli VioABCDE and VioABCDE-SD. The factors affecting violacein production, including temperature, pH, inoculum size, carbon and nitrogen source, precursor, and inducers were investigated. The violacein titer of VioABCDE-SD reached 107 mg/L in a two-stage fermentation process, representing a 454.4% increase over the original strain. The violacein operon from B9-8 provides a new microbial gene source for the analysis of the violacein synthesis mechanism, and the constructed engineering E. coli strains lay a foundation for the efficient and rapid synthesis of other natural products.

Key points

• The newly discovered violacein operon vioABCDE was identified in the genome of the extremophile Janthinobacterium sp. B9-8.

• The violacein synthesis pathway was reconstructed in E. coli using two compatible plasmids.

• A two-stage fermentation process was optimized for improved violacein accumulation.

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Data availability

All data generated or analyzed during this study are included in this published article (and its supplementary information file).

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Funding

This work was supported by the National Natural Science Foundation of China (21978136, 32060004), the National Natural Science Foundation of China Youth Fund (31922070), and the Natural Science Foundation of Jiangsu Province (BK20211268, BK20180038).

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Authors and Affiliations

  1. School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210046, Jiangsu Province, China

    Xian Xu, Bangmian Du & Chunyan Huang

  2. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, 211816, Jiangsu Province, China

    Xiaoting Chu

  3. School of Chemical Engineering, Yangzhou Polytechnic Institute, Yangzhou, 225127, Jiangsu Province, China

    Chengjia Xie

  4. Institute of Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi, 830091, Xinjiang Uyghur Autonomous Region, People’s Republic of China

    Zhidong Zhang

  5. College of Food Science and Light Industry, Nanjing Tech University, Nanjing, 211816, Jiangsu Province, China

    Ling Jiang

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  1. Xian Xu
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  2. Xiaoting Chu
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  3. Bangmian Du
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  4. Chunyan Huang
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Contributions

X. X. and L. J. designed the experiments. X. X., X. C., B. D., and C. H. performed the research. C. X. and Z. Z. collected and analyzed the data. Z. Z. and L. J. supervised the research. X. X. and X. C. wrote the paper. C. X., Z. Z., and L. J. polished the paper. All authors read and approved the manuscript.

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Correspondence to Xian Xu.

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Xu, X., Chu, X., Du, B. et al. Functional characterization of a novel violacein biosynthesis operon from Janthinobacterium sp. B9-8. Appl Microbiol Biotechnol 106, 2903–2916 (2022). https://doi.org/10.1007/s00253-022-11929-8

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