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A kind of marine bacillus and the polypeptide with anti-tumor activity produced by it

A marine bacillus and tumor technology, applied in the field of peptides with anti-tumor activity, can solve the problem of less research on anti-tumor lead compounds, achieve good research and application value, good acid-base and thermal stability, and low cytotoxicity Effect

Inactive Publication Date: 2014-10-29
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are relatively few studies on anti-tumor lead compounds derived from marine bacteria, and in-depth research in this area remains to be done.

Method used

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  • A kind of marine bacillus and the polypeptide with anti-tumor activity produced by it
  • A kind of marine bacillus and the polypeptide with anti-tumor activity produced by it
  • A kind of marine bacillus and the polypeptide with anti-tumor activity produced by it

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Screening and Isolation Process of Marine Bacillus N16

[0036] (1) Primary screening of activity

[0037] Seawater samples were selectively isolated and cultured with different media such as seawater basal medium 2216E and beef extract peptone medium NRG, and a total of 47 strains of single bacteria were obtained. Preliminary screening of the anti-tumor activity of the 47 microbial fermentation products showed that one strain had an inhibition rate of more than 60% on liver cancer cell BEL-7402; and a total of 1 strain had an inhibition rate of more than 50-60%.

[0038] (2) Strain re-screening

[0039] The proliferation inhibitory effect of the fermentation liquid of the two active strains above on other tumor cells, including human breast cancer cell MCF-7, human glioma cell U251 and human non-small cell lung cancer cell A549, was determined. The results showed that the fermentation liquid of one of the strains N16 had good cell proliferation inhibitory effect on t...

Embodiment 2

[0041] Morphological and Physiological and Biochemical Characteristics of Marine Bacillus N16

[0042] (1) Morphological characteristics

[0043] The strains were Gram-stained and found to be Gram-positive bacteria. Strain N16 has two important characteristics of Bacillus genus, bacillus morphology and spore-forming ability. It was cultivated by plate insert method, and the shape of strain N16 was observed by scanning electron microscope. It was 0.8-2.0 μm long and 0.2-0.6 μm wide ( figure 1 ). After cultured on seawater basal medium 2216E at 25°C for 24 hours, the colony is oval, entire, flat, with a diameter of 0.5 to 1.5 mm, smooth and matte, with a mucus-like appearance and milky white.

[0044] 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.

[0045] (2) Physiological and biochemical reaction characteristics

[0046] Mainly according to the methods of "General Bacter...

Embodiment 3

[0048] A small amount of bacterial genomic DNA was prepared, 16S rDNA was amplified by PCR, and the PCR product was detected as a single band by agarose gel electrophoresis, and then sent directly to Shanghai Sangon for sequencing. Test two reactions, test pass. The full length of the sequence is 1542bp, and the 16S rDNA sequence is shown in the sequence list.

[0049] The 16SrDNA sequence was compared with BLAST on the NCBI website, and the homologous sequence search was carried out. The results showed that the bacterium was Bacillus, named marine Bacillus sp.N16 (Bacillus sp.N16). The marine Bacillus N16 was preserved in the Wuhan Type Culture Collection Center of China on March 4, 2013, with the preservation number CCTCC M2013063. The 16S rDNA gene sequence has different mutation rates at certain sites, but in species, genus It shows the conservation of structure and function horizontally. It is known as "bacterial fossil" and is a timer for the history of biological evolu...

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Abstract

The invention relates to a marine bacillus and its polypeptide with antitumor activity. The strain is preserved at China Center for Type Culture Collection in Wuhan, and its preservation number is CCTCC M 2013063. A novel marine bacillus polypeptide with anti-tumor activity is obtained from a fermentation product of the strain. By means of an MTT method, the invention finds that the polypeptide has an obvious proliferation inhibition effect on human hepatoma carcinoma cell BEL-7402, human breast cancer cell MCF-7, human glioma cell U251, human non-small-cell lung cancer cell A549 and other tumor cells, has relatively small cytotoxicity on human fibroblast HFL1, and can be applied in drugs treating human liver cancer, gliomas, lung cancer and breast cancer.

Description

technical field [0001] The invention belongs to the field of marine microorganisms, and in particular relates to a marine bacillus producing anti-tumor active polypeptides and the produced anti-tumor active polypeptides. Background technique [0002] In today's society, malignant tumors seriously threaten human life and health, and are the number one killer of human society. Clinically, the treatment of malignant tumors mostly adopts comprehensive therapy, which combines surgical resection, radiotherapy, chemotherapy and immunotherapy. Although the antitumor drugs currently used have certain curative effects on most tumors, there are still problems such as low therapeutic efficiency, poor selectivity, obvious toxic and side effects, and easy generation of cell drug resistance. Therefore, finding anti-tumor drugs with high efficiency, low toxicity and clear targets is still a research hotspot in the field of biological and medical research. [0003] Due to the unique geogra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C07K7/06C12P21/02C07K1/36C07K1/34C07K1/18C07K1/16A61K38/08A61P35/00C12R1/07
Inventor 郑兰红孙谧郑媛王伟盛军纪晓峰郝建华
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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