Bacillus sp.N11-8 and active component thereof having antitumor action
A marine bacillus, anti-tumor activity technology, applied in the direction of anti-tumor drugs, medical preparations containing active ingredients, bacteria, etc., can solve the problems of difficult drug sources, difficult large-scale preparation, and low content
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Embodiment 1
[0029] Isolation of strains and primary screening of activity
[0030] Chase's medium Cha (for cultivating mold), seawater basal medium 2216E, beef extract peptone medium NRG (for cultivating bacteria), Gao's No. 1 medium G1 (for cultivating actinomycetes) and Martin's medium MD (for cultivating Fungi) and other different media were used to selectively isolate and cultivate Antarctic seawater samples, and a total of 50 single bacteria were obtained. Preliminary screening of the anti-tumor activity of these 50 microbial fermentation products showed that one strain had an inhibition rate of more than 60% against liver cancer cell BEL-7402, and the proportion of active strains was 2%; 2 strains; 5 strains with an inhibition rate of more than 30%; the remaining strains have almost no inhibitory effect on tumor cells.
Embodiment 2
[0032] Inhibitory Effects of Bacterial Strains on Different Tumor Cells
[0033] The three active strains with an inhibition rate of more than 50% were screened again, using human colon cancer cell HCT-116, human breast cancer cell MCF-7, pancreatic cancer cell P28, human non-small cell lung cancer cell A549 and human glioma Six strains of tumor cells such as cell U251 were used as targets, and the SRB method was used for multi-target screening, and it was found that the fermentation broth of Antarctic bacteria N11-8 (No. All of them have good proliferation inhibitory effect, and the inhibition rate of MCF-7 is 76.2%. As in Table 1:
[0034] Table 1 Inhibition rate of Antarctic bacteria N11-8 on different tumor cells
[0035]
Embodiment 3
[0037] Morphological and Physiological and Biochemical Characteristics of Strain N11-8
[0038] (1) Morphological characteristics
[0039] Gram staining results showed that the strain N11-8 was a Gram-negative bacteria. Strain N11-8 has two important characteristics of Bacillus, which are bacillus morphology and spore-forming ability. It is 0.5-2.5 μm long, 0.5-0.8 μm wide, and oval spores are terminal ( figure 1 ). After being cultured on beef extract peptone NRG medium for 24 hours at 25°C, the colonies are round, entire, flat, with a diameter of 0.8 to 2.1 mm, smooth and shiny, with a mucus-like appearance, creamy texture, and white translucent.
[0040] The bacterium is an aerobic bacterium and grows well when the pH value is 7.5, the culture temperature is 25°C, and the medium contains 6% (w / v) NaCl.
[0041] (2) Physiological and biochemical reaction characteristics
[0042] Mainly according to the gold standard for bacterial identification (API), use the API20E and AP...
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