Applied Microbiology and Biotechnology

, Volume 94, Issue 6, pp 1521–1532 | Cite as

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

  • Pei-xia Jiang
  • Hai-sheng Wang
  • Su Xiao
  • Ming-yue Fang
  • Rui-ping Zhang
  • Shu-ying He
  • Kai Lou
  • Xin-Hui Xing
Biotechnological products and process engineering

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.

Keywords

Violacein Deoxyviolacein Biosynthesis Heterologous expression Pathway redesign 

Supplementary material

253_2012_3960_MOESM1_ESM.docx (1 mb)
ESM 1(DOCX 1028 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Pei-xia Jiang
    • 1
    • 2
  • Hai-sheng Wang
    • 3
  • Su Xiao
    • 2
  • Ming-yue Fang
    • 2
  • Rui-ping Zhang
    • 2
  • Shu-ying He
    • 4
  • Kai Lou
    • 5
  • Xin-Hui Xing
    • 2
  1. 1.Department of Industrial Microbiology and Biotechnology, Institute of MicrobiologyChinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.Institute of Biochemical Engineering, Department of Chemical EngineeringTsinghua UniversityBeijingPeople’s Republic of China
  3. 3.Graduate School of Chinese Academy of Agricultural SciencesBeijingPeople’s Republic of China
  4. 4.School of Life Science and TechnologyChina Pharmaceutical UniversityNanjingPeople’s Republic of China
  5. 5.Institute of MicrobiologyXinjiang Academy of Agricultural SciencesÜrümqiPeople’s Republic of China

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