Deep-sea streptomyces, Tianyamycin series compounds and application of series compounds

A technology of streptomyces and compounds, applied in the field of biomedicine, can solve problems such as difficulty in discovering new antibiotics

Active Publication Date: 2021-08-27
INST OF DEEP SEA SCI & ENG CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] With the full development and utilization of terrestrial microbial resources, the process of discovering new natural products has become a bottleneck, making it increasingly difficult to discover new antibiotics

Method used

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  • Deep-sea streptomyces, Tianyamycin series compounds and application of series compounds
  • Deep-sea streptomyces, Tianyamycin series compounds and application of series compounds
  • Deep-sea streptomyces, Tianyamycin series compounds and application of series compounds

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preparation example Construction

[0092] The third object of the present invention: to provide the preparation method of compound 1-4 as shown in formula I. It is characterized in that it includes the following steps.

[0093] a, preparing a fermentation culture of Streptomyces sp.LDBS7850;

[0094] b. Separating the fermentation culture to obtain bacteria and fermentation broth. The fermentation broth was extracted three times with an equal volume of ethyl acetate:n-hexane (1:1) mixture, and the collected extract was rotary evaporated to dryness and stored in a -20°C refrigerator. Add 2-4 times the volume of the bacteria to the methanol-acetone (1:4) mixture, sonicate for 20 minutes, filter and collect the extract; repeat the extraction of the bacteria 3 times; pour the collected 1500mL extract into the rotary steamer , concentrated in vacuum at 34°C and 150-300hPa into a rotary evaporator with a solid-to-liquid ratio of 1:2, added ultrapure water to fully dissolve; then, added an equal volume of ethyl acet...

Embodiment 1

[0106] Example 1: Culture of Streptomyces sp.LDBS7850 and identification of 16s rRNA gene

[0107] 1. Isolation and cultivation of Streptomyces sp.LDBS7850 strain.

[0108] 1. Solid culture of Streptomyces sp.LDBS7850:

[0109]1.1 The Streptomyces sp.LDBS7850 strain was isolated from the sediments at a depth of 7850m in the Mariana Trench, and the strain was preserved in Marine Broth2216 glycerol medium.

[0110] 2. 16s rRNA gene sequence analysis of Streptomyces sp.LDBS7850 strain.

[0111] 2.1 Extraction of Streptomyces sp.LDBS7850 strain genomic DNA and PCR amplification, sequencing and species identification of 16s rRNA gene. The genome of Streptomyces sp.LDBS7850 was extracted using a DNA extraction kit (TIANGEN) and part of it was used as a template. PCR amplification was performed using 16S universal primers and Taq enzyme from TAKARA Company at an annealing temperature of 55°C. The amplified PCR product was sent to Huada Gene Company for bidirectional sequencing. T...

Embodiment 2

[0123] Example 2: Using Streptomyces sp.LDBS7850 to produce Tianyamycin series compounds.

[0124] 1. Fermentation and extraction of Streptomyces sp.LDBS7850 and extraction and separation of Tianyamycin.

[0125] (1) Fermentation: Take out the Streptomyces sp.LDBS7850 glycerin tube from the refrigerator at -80°C, inoculate it on the Marine Broth2216 plate with the inoculation loop method, and place it in a constant temperature incubator at 30°C for 4-5 days until a large number of white spores grow on the plate. Use an inoculation loop to pick a small amount of spores into 50mL of SPY medium (10g soluble starch; 5g yeast extract; 3g tryptone; 21g sea salt; 1LddH2O; pH7.2-7.6), and culture on a constant temperature shaker for 3 days (180rpm, 34° C.) to obtain seed solution. 50 mL of the seed solution was inoculated into 1 L of SPY fermentation medium, and cultured on a constant temperature shaker (180 rpm, 34° C.) for 7 days.

[0126] (2) Extraction: 6 liters of LDBS7850 ferm...

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Abstract

The invention relates to the field of biological medicine, in particular to streptomyces abyssinus, Tianyamycin series compounds and application of the compounds. According to the invention, four compounds (compounds 1-4) are separated from a fermentation culture Streptomyces sp.LDBS7850 with antipathogenic activity derived from a Mariana trench sediment, and the compounds 1-4 are subjected to structural identification to be new tetramic acid compounds named as Tianyamycin A-D. The four compounds show very strong activity of inhibiting MRSA (Methicillin-Resistant Staphylococcus Aureus). In addition, the four compounds have different degrees of inhibition effects on methicillin-sensitive staphylococcus aureus and enterococcus faecalis.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to deep-sea streptomyces, Tianyamycin series compounds and applications thereof. Background technique [0002] Since the British scientist Alexander Fleming (Alexander.Fleming) first discovered antibiotics—penicillin in 1928, antibiotics have played a huge role in human infectious diseases. However, people's over-reliance on and overuse of antibiotics has led to more and more types of drug-resistant and multidrug-resistant bacteria. Drug-resistant bacterial infection has become a public health problem that cannot be ignored, posing a huge threat to human health. The World Health Organization (WHO) has repeatedly called for and encouraged the research and development of new antibiotics. Therefore, the society urgently needs some new antibiotics with unique mechanisms. [0003] With the full development and utilization of terrestrial microbial resources, the process of discovering new nat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/16C07D405/06A61K31/4025A61P31/04C12R1/465
CPCC12P17/16C07D405/06A61P31/04
Inventor 韩壮王佳伟翟吉安高兆明
Owner INST OF DEEP SEA SCI & ENG CHINESE ACADEMY OF SCI
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