Marine aspergillus versicolor as well as isolated culture method and application thereof
A kind of Aspergillus versicolor and marine technology, applied in the field of microorganisms, can solve the problems of insufficient natural drug sources and drug resistance of cancer cells, and achieve good application potential and promotion prospects, good application value, and easy-to-achieve effects
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Embodiment 1
[0049] Isolate and cultivate marine Aspergillus versicolor strains by the method provided by the invention, the specific operation steps are as follows:
[0050] S1. Sample collection and processing: Collect 3 to 5 samples of scleractinia from the Nansha Islands, put them into sterile plastic bags, and send them to the laboratory as soon as possible under freezing conditions, and then put the scleractinia samples in sterile seawater Rinse three times to remove loosely attached microorganisms, and take 15-20g of the washed sample, cut into a volume of about 1cm 3 small pieces and crushed in a mortar;
[0051] S2. Sample inoculation: add the sample ground in step S1 into sterile seawater and sterile sand with a volume ratio of 2:1 and mix evenly. After the homogenate is diluted 10 times with sterile seawater, measure 0.1mL and inoculated onto pre-prepared agar plates, this step set up 3 parallel groups;
[0052] S3. Isolation and cultivation of bacterial strains: Place the ino...
Embodiment 2
[0058] The exopolysaccharide is prepared by isolating and cultivating Aspergillus versicolor SCAU141 in Example 1 of the present invention, and the preparation method comprises the following steps:
[0059] 1. Inoculate the marine Aspergillus versicolor strain in the liquid medium (the composition of the liquid medium used is: maltose 20g / L, glucose 10g / L, mannitol 20g / L, sodium glutamate 10g / L, magnesium sulfate heptahydrate 0.3g / L, potassium dihydrogen phosphate 0.5g / L, corn steep liquor 1g / L, yeast extract 3g / L and sea salt 30g / L, with a pH value of 7.5), cultured and fermented for 7 days at 28°C and 130rpm to obtain Marine Aspergillus versicolor fermentation broth;
[0060] 2. Filter the Aspergillus versicolor fermented liquid obtained in step 1 to remove the thalline, collect the filtrate, concentrate under reduced pressure, add four times the volume of absolute ethanol to the concentrated solution, and place the obtained mixed solution overnight at low temperature;
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experiment example 1
[0086] In order to verify the anticancer effect of the exopolysaccharide produced by Aspergillus versicolor SCAU141 of the present invention, the extracellular polysaccharide prepared in Example 2 was used to carry out the cancer cell growth inhibition experiment. In order to reflect the experimental results, the cell The tumor suppressor activity of exopolysaccharides is characterized by the absorbance of ATP measured by luminescence method, because the ATP cell viability test solution can sensitively and linearly quantify 50 to 50,000 living cells in culture in a 96-well plate, which is a fast, effective and accurate detection method.
[0087] The tumor suppression experiments of extracellular polysaccharides were performed on lung cancer A549 cells, liver cancer Hep-G2 cells, and colon cancer HT-29 cells. The experimental methods are as follows:
[0088] 1. The culture conditions of cancer cells used in the experiment:
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