Streptomyces atratus and application of cyclic peptide compounds with same to preparing mycobacterium tuberculosis resistant medicines

A technology of marine streptomyces and mycobacteria, applied to cyclic peptide compounds and their application in the preparation of anti-tuberculosis drugs, the field of marine streptomyces

Active Publication Date: 2017-01-04
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to its unique mechanism of action, the drug is limited to the trea...

Method used

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  • Streptomyces atratus and application of cyclic peptide compounds with same to preparing mycobacterium tuberculosis resistant medicines
  • Streptomyces atratus and application of cyclic peptide compounds with same to preparing mycobacterium tuberculosis resistant medicines
  • Streptomyces atratus and application of cyclic peptide compounds with same to preparing mycobacterium tuberculosis resistant medicines

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Experimental program
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Effect test

Embodiment 1

[0041] Culture of Streptomyces atratus SCSIO Zh16, 16s rRNA gene identification, fermentation detection and HPLC analysis of metabolites.

[0042] 1. Fermentation detection, HPLC analysis and 16s rRNA gene sequence analysis of S.atratus SCSIO Zh16 strain

[0043] 1. Solid culture of S.atratus SCSIO Zh16:

[0044] 1.1 Marine Streptomyces S.atratus SCSIO Zh16 was isolated from deep-sea sediments in the South China Sea. The strain was stored on the slope of ISP-2 medium. The composition of ISP-2 medium was: yeast extract powder 4g, glucose 4g, malt extract powder 10g , coarse sea salt 30g, agar powder 20g, water 1000mL, pH 7.2-7.4.

[0045]1.2 S.atratus SCSIO Zh16 strain genomic DNA extraction and 16s rRNA gene PCR amplification, sequencing and NCBI BLAST comparative analysis. Genomic DNA of S.atratus SCSIO Zh16 was extracted by microwave method and a part was taken as a template. Using 16S universal primers, rTaqase from Trans Gene Company was used for PCR amplification at 58°...

Embodiment 2

[0067] Production of cyclic peptide compounds using S.atratus SCSIO Zh16.

[0068] 1. Large-scale fermentation of S.atratus SCSIO Zh16

[0069] (1) Configuration of seed medium and fermentation medium

[0070] The configuration of the seed medium (Am2ab): soybean powder 10g, soluble starch 10g, glucose 40g, peptone 4g, yeast extract powder 4g, MgSO4.7H 2 O 1g, KH 2 PO 4 1g, NaCl 8g, CaCO 3 4g, 60g sea salt, add 2L water, pH 7.2–7.4, evenly distribute in 40 250mL Erlenmeyer flasks, and sterilize at 121°C for 25 minutes to obtain sterilized seed culture medium.

[0071] Preparation of fermentation medium (Am3): 5g of soybean powder, 15g of bacteriological peptone, 15g of soluble starch, 15g of glycerol, CaCO 3 2g, 30g sea salt, 1L water, pH 7.2–7.4. A total of 16.2 liters were prepared, distributed in 72 conical flasks of 1000 mL, and sterilized at 121° C. for 30 minutes to obtain sterilized fermentation medium.

[0072] (2) Fermentation culture:

[0073] The strain o...

Embodiment 3

[0088] Antibacterial activity test analysis of compounds 1-6 shown in formula II against mycobacteria.

[0089] Compounds 1-6 were tested against mycobacterium M. smegmatis mc by broth doubling dilution method 2 155 and M. tuberculosis H37R V antibacterial activity. Among them, M.smegmatis mc 2 155 antibacterial activity test operation steps are briefly described as follows: 1) put M.smegmatis mc 2 155 was resuscitated for 36 hours, and diluted 1000 times when used as the bacterial solution to be tested; 2) Compound 1-6 was dissolved in DMSO to prepare a stock solution with a concentration of 3200 μg / mL; 3) The drug stock solution was mixed with 7H9 in a certain ratio Broth (adding 0.2% glycerol, 0.05% Tween 80) was diluted, and finally diluted to a final concentration of 256-0.125 μg / mL, and the volume of drug broth in each well was 100 μL; Add 100 μL of diluted bacterial solution to each well, so that the final concentration of each well in each row is 128-0.0625 μg / m...

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Abstract

The invention discloses streptomyces atratus and application of cyclic peptide compounds with the same to preparing mycobacterium tuberculosis resistant medicines. A structural formula of the cyclic peptide compounds is shown. A preservation number of the streptomyces atratus SCSIO Zh16 is CGMCC No.12198. The streptomyces atratus and the application have the advantages that the six cyclic peptide compounds are obtained from fermentation cultivation substances of the streptomyces atratus SCSIO Zh16 by means of separation, the cyclic peptide compound 6 is high in mycobacterium tuberculosis resistant activity, obvious effects of inhibiting mycobacterium tuberculosis can be realized by the cyclic peptide compound, accordingly, the cyclic peptide compounds can be used for preparing anti-tuberculosis medicines and can be used for treating tuberculosis, alternative compounds can be provided for developing novel anti-tuberculosis medicines, and the streptomyces atratus and the application have important significance on developing marine medicinal materials in China.

Description

[0001] Technical field: [0002] The invention belongs to the field of natural products, and in particular relates to a marine streptomyces, a cyclic peptide compound and an application thereof in preparing anti-tuberculosis mycobacterium drugs. [0003] Background technique: [0004] Mycobacterium is a general term for a group of pathogens, the most notorious of which are Mycobacterium tuberculosis, Mycobacterium leprae and Brucella. It has brought serious adverse effects on the development of the industry, and has become a social problem that endangers public health security and a heavy economic burden on social development. Worldwide, tuberculosis is already the leading cause of death from infectious diseases, surpassing the previous death rate from human immunodeficiency virus (HIV). According to the annual report on tuberculosis of the World Health Organization in 2015, there are about 2 billion people in the world (1 / 3 of the world's population) who are latently infected...

Claims

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Application Information

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IPC IPC(8): C07K7/64C12N1/20C12P21/04A61K38/12A61P31/06C12R1/465
CPCA61K38/00C07K7/64C12N1/205C12R2001/465
Inventor 鞠建华张天宇马俊英刘永志
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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