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Polar maritime actinomycete AFN1007 generating antineoplastic activity material

An anti-tumor activity, marine actinomycete technology, applied in the field of anti-tumor activity, can solve the problem of difficult to find effective therapeutic drugs for cancer

Inactive Publication Date: 2008-12-24
CHINA PHARM UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the other hand, more and more infectious bacteria have gradually developed resistance to traditional antibiotics, and cancer, one of the three major diseases threatening human health, is also difficult to find effective drugs for treatment, thus forcing biologists to look for new natural organisms active substances to solve current problems

Method used

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  • Polar maritime actinomycete AFN1007 generating antineoplastic activity material
  • Polar maritime actinomycete AFN1007 generating antineoplastic activity material
  • Polar maritime actinomycete AFN1007 generating antineoplastic activity material

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] 1. AFN1007 was cultured on Gaoshi Synthetic Medium No. 1 by the insert method, and the mycelium was observed under an Olympus BH-2 optical microscope. The spore chains showed a regular spiral shape, which belonged to the genus Streptomyces.

[0012] 2. The marine strain AFN1007 was cultured according to the medium formula recommended by the International Streptomyces Program (ISP) and Waksman, and the culture characteristics and physiological and biochemical characteristics were observed.

[0013] Strain AFN1007 grew well on 8 media. Most of the hyphae in the base were white or yellow, and the aerial hyphae were all white. Most of the spores were white and secreted red and gray-green soluble pigments (Table 1).

[0014] Table 1. Culture characteristics of arctic actinomycetes AFN1007

[0015]

[0016] Actinomycetes AFN1007 can grow on the medium containing NaCl in the range of 5‰-80‰. Seven carbon sources of glucose, sucrose, lactose, xylose, raffinose, mannitol, an...

Embodiment 2

[0030] 1. Fermentation experiment of polar marine actinomycete AFN1007: the medium components used are: soybean powder 1%, soluble starch 1%, KNO 3 0.1%, MgSO 4 0.05%, FeSO 4 0.001%, NaCl 0.05%, natural seawater, pH7.2-7.4. Inoculate the spores of the bacterial strain AFN1007 into a 250mL Erlenmeyer flask (containing 50mL medium), and cultivate it on a shaker at 200r / min at 15°C for 2 days as a seed solution; , cultivated under the same conditions for 10 days, sampled 10mL every 24 hours, then centrifuged at 4,000r / min for 10min, took the supernatant, measured the pH value, then sterilized with a 0.22μm filter membrane, and stored in a refrigerator at 4°C for testing Anti-tumor activity; Bacteria balls were baked at 90°C until the weight did not change, and the dry weight of the bacteria was measured.

[0031] 2. Antitumor activity detection method of fermentation broth: KB oral epithelial cancer cells in logarithmic growth phase were digested with 0.25% trypsin to make ...

Embodiment 3

[0036] According to the above fermentation experiment results, the fermentation of the fermentation strain AFN1007 was completed on the 7th day, and the fermentation broth was collected and diluted according to the ratio of Example 2 to detect the effects of each sample on KB oral epithelial cancer cells, gastric cancer cells BGC 803, liver cancer cells Hep G2 and cervical cancer cells. Antitumor effect of cancer cell Hela. It can be seen from Table 5 that the fermentation broth has inhibitory effects on the four tumor cells, among which the effect on KB oral epithelial cancer cells is the strongest, IC 50 was 2235.6±359.8, but the effect on Hep G2 liver cancer cells was weak, IC 50 It is 1723.4±256.3.

[0037] Table 5. Effect of AFN1007 on proliferation of 4 kinds of tumor cells in vitro

[0038]

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Abstract

The invention relates to a method for verifying and fermenting polar marine ray fungi AFN1007 producing antitumor active substance, and the research on the antitumor activity of the zymotic fluid thereof, belonging to the microbial medicine field. The strain AFN1007 is verified to be a novel kind of Streptomyces by the traditional classification and the analysis of 16S rDNA gene of the AFN1007. The compositions of a culture medium by percentage are: 1 percent of soybean flour, 1 percent of tragantine, 0.1 percent of KNO3, 0.05 percent of MgSO4, 0.001 percent of FeSO4, 0.05 percent of NaCl and natural seawater with a pH value between 7.2 and 7.4. The strain is fermented by using the culture medium. The MTT method shows that the zymotic fluid has strong antitumor action on four tumor cells of KB, BGC 803, Hep G2 and Hela and an antitumor compound with a novel structure is expected to be found in the zymotic fluid.

Description

technical field [0001] The invention relates to the identification and fermentation of a polar marine actinomycete AFN1007 producing anti-tumor active substances and the anti-tumor activity of its fermented liquid, belonging to the field of microbial medicines. Background technique [0002] In 1929, since Fleming in the United Kingdom discovered that microorganisms produced penicillin, it has been proved that microorganisms (including bacteria, actinomycetes, fungi, etc.) are a rich source of biologically active substances. Microorganisms can produce antibacterial, antitumor, antiviral and anticardiovascular bioactive substances. According to statistics, more than 8,000 natural products with antibacterial and antitumor activities have been discovered from microorganisms. However, the number of newly discovered bioactive substances from terrestrial microorganisms is decreasing, and most of them are known compounds. On the other hand, more and more infectious bacteria have g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12P1/04A61P35/00C12R1/465
Inventor 奚涛那广水张婷沈晶
Owner CHINA PHARM UNIV
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