Violacein and/or deoxyviolacein for treating colon cancer
A purple bacteriocin, deoxyviolet technology, applied in the field of purple bacteriocin and/or deoxyviolet bacterin for the treatment of colon cancer, which can solve problems such as low yield
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Embodiment 1
[0054] Example 1 The preparation of violacein and deoxyviobacterin
[0055] The violacein adopts the fermentation strain of Citrobacter freundii (Citrobacter freundii) (the public of this bacterial strain can obtain from Tsinghua University), and the pCom10Vio plasmid is contained in this bacterial strain (the non-patent literature recording this plasmid construction method is Jiang, P.X., Wang H.S., Zhang C., et al., Reconstruction of the violaceinbiosynthetic pathway from Duganella sp.B2 in different heterologous hosts.Applied Microbiology and Biotechnology, 2009,86(4):1077-88), the Freund's lemon with pCom10Vio plasmid The construction of Citrobacter freundii can be found in Zhang Ruiping, Jiang Peixia, Li Chun, et al. The optimization of process conditions for the synthesis of violacein by recombinant Citrobacter freundii [J]. Chemical Journal, 2010, 61(6): 1495-1505 .
[0056] The above-mentioned Citrobacter freundii with pCom10Vio plasmid was inoculated in 200mL ferment...
Embodiment 2
[0062] Example 2 Effects of violacein and / or deoxyviobacterin on the inhibition of HT29 cell proliferation
[0063] The detection indicators and methods used are as follows. The growth inhibition rate of HT29 cells was determined by MTT method, and the experiments were carried out according to the following groups.
[0064] Negative control group: routinely culture HT29 cells with McCoy’s 5A medium;
[0065] Positive control group: cells cultured in McCoy’s5A medium containing 5-fluorouracil (5-Fu) at a mass concentration of 5.000 μg / mL (also called 5FU group);
[0066] Experimental group 1 (viobacterin group (also known as Vio group)): cells were cultured in McCoy’s 5A medium containing violacein at a mass concentration of 0.125, 0.250, 0.500, 1.000, 2.000, 4.000 μg / mL.
[0067] Experimental group 2 (deoxyviolet violacein group (also known as dVio group)): cells were cultured in McCoy’s 5A medium containing deoxyviolet violacein at a mass concentration of 0.4, 0.8, 1.6, 3.2...
Embodiment 3
[0079] Example 3 Effects of violacein and / or deoxyviobacterin on HT29 cell cycle
[0080] Use the above-mentioned flow cytometer to detect the cell cycle, take the HT29 cells in the logarithmic growth phase and adjust the cell concentration to 10 5 cells / mL, inoculated in a 6-well plate, and inoculated 2 mL of cell suspension in each well. Discard the culture medium after cell culture for 24 hours, add culture medium containing violacein with a mass concentration of 0.000 (negative control group), 0.125, 0.500, and 2.000 μg / mL to continue culturing for 24 hours, then remove the culture medium, or add violacein-containing medium The mass concentration is 0 (negative control group), and the culture medium of 0.8, 3.2, and 12.8 μg / mL is continuously cultured for 48 h and then the culture solution is removed (in addition, the mass concentration of 5.000 μg of 5-fluorouracil (5-Fu) is also used. After the McCoy's 5A culture medium of / mL continued to cultivate for 48h, remove the ...
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