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Pathway redesign for deoxyviolacein biosynthesis in Citrobacter freundii and characterization of this pigment

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Abstract

Violacein (Vio) is an important purple pigment with many potential bioactivities. Deoxyviolacein, a structural analog of Vio, is always synthesized in low concentrations with Vio in wild-type bacteria. Due to deoxyviolacein’s low production and difficulties in isolation and purification, little has been learned regarding its function and potential applications. This study was the first effort in developing a stable and efficient biosynthetic system for producing pure deoxyviolacein. A recombinant plasmid with vioabce genes was constructed by splicing using an overlapping extension-polymerase chain reaction, based on the Vio-synthesizing gene cluster of vioabcde, originating from Duganella sp. B2, and was introduced into Citrobacter freundii. With the viod gene disrupted in the Vio synthetic pathway, Vio production was completely abolished and the recombinant C. freundii synthesized only deoxyviolacein. Interestingly, vioe gene expression was strongly stimulated in the viod-deleted recombinant strain, indicating that viod disruptions could potentially induce polar effects upon the downstream vioe gene within this small operon. Deoxyviolacein production by this strain reached 1.9 g/L in shaker flasks. The product exhibited significant acid/alkali and UV resistance as well as significant inhibition of hepatocellular carcinoma cell proliferation at low concentrations of 0.1–1 μM. These physical characteristics and antitumor activities of deoxyviolacein contribute to illuminating its potential applications.

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Acknowledgments

This work was supported in part by the National Science Fund of China (Grant no. 21006058), a China Postdoctoral Science Foundation funded project (Grant no. 20080430367), a Basic Research Fund of CAAS (0042009001), and a Xinjiang-supporting project by Science and Technology (Grant no. 200991132).

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Authors and Affiliations

  1. Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    Pei-xia Jiang

  2. Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing, 100084, People’s Republic of China

    Pei-xia Jiang, Su Xiao, Ming-yue Fang, Rui-ping Zhang & Xin-Hui Xing

  3. Graduate School of Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Hai-sheng Wang

  4. School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009, People’s Republic of China

    Shu-ying He

  5. Institute of Microbiology, Xinjiang Academy of Agricultural Sciences, Ürümqi, 830091, People’s Republic of China

    Kai Lou

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  1. Pei-xia Jiang
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  2. Hai-sheng Wang
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  8. Xin-Hui Xing
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Correspondence to Xin-Hui Xing.

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Jiang, Px., Wang, Hs., Xiao, S. et al. Pathway redesign for deoxyviolacein biosynthesis in Citrobacter freundii and characterization of this pigment. Appl Microbiol Biotechnol 94, 1521–1532 (2012). https://doi.org/10.1007/s00253-012-3960-0

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  • DOI: https://doi.org/10.1007/s00253-012-3960-0

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