violacein and/or deoxyviobacterin for promoting wound healing and blood clotting
A purple bacteriocin, promoting technology, applied in blood diseases, drug combinations, medical preparations containing active ingredients, etc., can solve problems such as low yield
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Embodiment 1
[0072] The preparation of embodiment 1 violacein and deoxy violacein
[0073] 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.B2in different heterologous hosts.Applied Microbiology and Biotechnology, 2009,86(4):1077-88), the Freund's citric acid with pCom10Vio plasmid The construction of Citrobacter freundii can be found in the description in Zhang Ruiping, Jiang Peixia, Li Chun, et al. Optimization of process conditions for the synthesis of violacein by recombinant Citrobacter freundii[J].
[0074] The above-mentioned Citrobacter freundii with pCom10Vio plasmid was inoculated in 200mL fermentation medium (beef extr...
Embodiment 2
[0082] Example 2 Effects of violacein and deoxyviobacterin on activated partial thrombin time (APTT)
[0083] Take 3.6mL of blood from the vein of the rex rabbit's ear, put it into a 4mL centrifuge tube containing 400μL of 0.109mol / L sodium citrate, mix it by inverting gently, centrifuge at 3000r / min for 15min, and take the supernatant as plasma for later use. Add 20 μL of the above-prepared violacein solution, deoxy violacein solution, vitamin K 1 solution, breviscapine solution, and negative control solution, and then add 100 μL of the above-mentioned spare plasma and 100 μL of APTT reagent pre-warmed at 37 °C, incubate at 37 °C for 5 min, then add 0.025 mol / L CaCl pre-warmed at 37 °C 2 The solution was 100 μL, and the coagulation time was recorded, which was the measured value of APTT.
[0084] Measurement results such as figure 1 As shown, among them, figure 1 The vertical axis of indicates APTT measurement value, that is, the result of activated partial thrombin time. ...
Embodiment 3
[0085] Example 3 Effects of violacein and deoxyviobacterin on prothrombin time (PT)
[0086] Use the same method as in Example 2 to obtain plasma for later use, and add 20 μL of the above-prepared violacein solution, deoxy violacein solution, and vitamin K to 5 test cups. 1 solution, breviscapine solution and negative control solution, and then add 100 μL of the above spare plasma and 200 μL of PT reagent pre-warmed at 37°C, record the coagulation time, which is the measured value of PT.
[0087] From the preliminary results of Example 2, violacein and deoxyviobacterin did not significantly shorten the PT compared with the negative control without adding violacein, and it is estimated that violacein and deoxyviolacein have little effect on the extrinsic blood coagulation pathway .
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