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Microbe composite polysaccharide for suppressing cancer cell and its preparation method

A technology of compound polysaccharide and microorganism, applied in the field of biomedicine

Inactive Publication Date: 2006-05-10
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few reports on the anti-tumor and immune-enhancing effects of microbial polysaccharides. We isolated and screened a strain of Penicillium oxalicum (Penicillium oxalicum) from the environment, and the preservation number: CGMCC No.0907. Polysaccharide has high activity against human promyelocytic leukemia cell line HL-60 and human liver cancer cell line Bel-7402

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1: the thalline culture part of Penicillium oxalicum CGMCC No.0907 bacterial strain

[0014] (1) Expanded culture of strains: weigh 15g of beet pulp and 8g of bran into a 500mL conical flask, add 50mL of tap water dissolved with 1.5g of ammonium sulfate, stir evenly, then add 25mL of water dissolved with 0.3g of disodium hydrogen phosphate , stir evenly, and sterilize at 115°C for 15 minutes. Inoculate the strains preserved on the slant, and culture statically at 30°C for 3-5 days. After the spores grow densely, store them in a refrigerator at 4°C for later use.

[0015] (2) Liquid culture: Weigh 1.0g of peptone, 0.6g of yeast extract, 1.0g of malt extract powder, 1.5g of glucose, 0.2g of sodium chloride, 0.1g of potassium dihydrogen phosphate, and disodium hydrogen phosphate into a 500mL Erlenmeyer flask 0.2g, add 100mL of water, and sterilize at 115°C for 15 minutes. Inoculation step (1) A small key for the strain, placed on a shaker, 160-180 rpm, 30°C sh...

Embodiment 2

[0017] Embodiment 2: extract complex polysaccharide from Penicillium oxalicum CGMCC No.0907 thalline (spore)

[0018] (1) Boiling water extraction liquid culture mycelia composite polysaccharide: add about 30 times the weight of deionized water to the wet mycelium obtained by liquid culture, boil and reflux for 2 hours, centrifuge, and collect the extract; Add water and boil under reflux for 1 hour, centrifuge to collect the extract; combine the two extracts, continue to boil and evaporate to concentrate until the total sugar content of the solution reaches 2%, add twice the volume of 95% ethanol, centrifuge, collect the precipitate, and then use 95% ethanol Wash the precipitate, centrifuge, collect the precipitate, and dry it naturally to obtain the complex polysaccharide.

[0019] (2) Combined extraction of solid-state cultured bacteria (spores) polysaccharides by boiling water and alkali: first extract the bacteria or spores obtained by solid-state culture with boiling wate...

Embodiment 3

[0020] Embodiment 3, antitumor experiment

[0021] IC 50 Determination. HL-60 cells and Bel-7402 cells in the logarithmic growth phase were taken respectively and added to 96-well culture plates, and samples of different dilutions were added at the same time, and 3 replicate wells were set for each concentration. After 3-5 days of culture, count with a hemocytometer. Plot the cell viability-drug concentration curve. The results showed that the median lethal concentration (IC) of the microbial complex polysaccharide to HL-60 cells 50 ) is 0.3~2.2μg / mL, when the concentration reaches 6.0μg / mL, the cells do not grow at all; the median lethal concentration (IC) for Bel-7402 cells 50 ) was 120-150 μg / mL, and when the concentration reached 300 μg / mL, the cells did not grow at all.

[0022] The biological material sample of the present invention is preserved in the General Microbiology Center of China Microbiological Strain Preservation Management Committee, address: No. 13, Nor...

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Abstract

The invention provides the microbe polysaccharides compounds of suppressing the activity of human promyelocytic leukemia cells (HL-60) and human hepatocellular carcinoma cell (Bel-7402), and also provides its preparation method, belonging to the biological medicine technology field. The invention comprises the following steps: adopting the Penixillium oxalicum strain (CGMCC No.0907), getting the thalline and spore by liquid and solid state culture method; extracting it by boiling-water, alkali, and boiling-water and alkali respectively, finally getting the microbe polysaccharides compounds. The half lethal concentration (IC50) of microbe polysaccharides compounds in HL-60 is 0.3-2.2ª–g / mL, and when the concentration is 6.0ª–g / mL, the HL-60 doesn't grow completely; the half lethal concentration (IC50) of microbe polysaccharides compounds in Bel-7402 is 120-150ª–g / mL, and when the concentration is 300ª–g / mL, the cell doesn't grow completely.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. technical background [0002] At least 650 polysaccharides from higher basidiomycetes (mushrooms) have anti-tumor and immune-enhancing activities (S.P. Wasser, Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. Appl Microbiol Biotechnol, 2002, 60: 258-274). A Chinese patent (publication number: CN 1454904A) discloses a method for preparing high-purity fungal polysaccharides. The fungi used are shiitake mushrooms, ganoderma lucidum and Agaricus blazei. A Chinese patent (authorized announcement number: CN 1142283C) discloses a method for preparing active polysaccharides by using biotechnology, which is prepared by mixed fermentation of yeast, ganoderma lucidum and shiitake mushrooms. Chinese patent (publication number: CN 1297994A) discloses a method for producing edible and medicinal fungal powder and fungal polysaccharides. Mushrooms, Flammulina velutipes and...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/715A61P35/00C12N1/14
Inventor 白志辉朱显峰张保国张洪勋
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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