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

Springer Science and Business Media LLC - Tập 94 - Trang 1521-1532 - 2012
Pei-xia Jiang1,2, Hai-sheng Wang3, Su Xiao2, Ming-yue Fang2, Rui-ping Zhang2, Shu-ying He4, Kai Lou5, Xin-Hui Xing2
1Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, People’s Republic of China
2Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing, People’s Republic of China
3Graduate School of Chinese Academy of Agricultural Sciences, Beijing, People’s Republic of China
4School of Life Science and Technology, China Pharmaceutical University, Nanjing, People’s Republic of China
5Institute of Microbiology, Xinjiang Academy of Agricultural Sciences, Ürümqi, People’s Republic of China

Tóm tắt

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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Springer Science and Business Media LLC

Tập 94

1521-1532

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