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New streptomyces strain as well as separation method and application thereof

A separation method, the technology of Streptomyces, applied in the field of microbial pharmacy, can solve problems such as human health hazards, antibiotic resistance, and increased resistance of pathogens, and achieve the effect of enriching the production capacity of natural products

Pending Publication Date: 2022-04-12
BEIFANG UNIV OF NATITIES +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the widespread use of antibiotics for a period of time, even local abuse, the drug resistance of pathogens has increased sharply, and even developed into super-resistant bacteria, such as Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Bao Pathogens such as Acinetobacter manninii, Pseudomonas aeruginosa, and Enterobacter
These pathogens are resistant to almost all antibiotics currently used clinically, causing serious harm to human health

Method used

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  • New streptomyces strain as well as separation method and application thereof
  • New streptomyces strain as well as separation method and application thereof
  • New streptomyces strain as well as separation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Isolation, identification and preservation of bacterial strains of the present invention

[0038] (1) Separation

[0039] Add 1 g of soil samples collected from Ningxia saline-alkali land (longitude 106°08', latitude 38°38') and appropriate amount of glass beads to 9 mL of sterile water, shake for 10 min, and let stand for 30 s to make a soil stock solution (at this time, the dilution is recorded as 10 -1 ). Take 1 mL of the supernatant of the soil stock solution and add it to a test tube filled with 9 mL of sterile water and mix well (the dilution is 10 at this time).-2 ), draw 1mL from this test tube and add it to another test tube filled with 9mL sterile water, mix well, and make a dilution by analogy (the dilution is 10 -4 ).

[0040] Use a pipette gun to draw 100 μL of diluents of different dilutions and evenly spread them on the Gaoshi No. 1 solid medium added with 100 mg / L sterile potassium dichromate, culture at 28°C for 5-7 days, and observe th...

Embodiment 2

[0052] Example 2: Determination of the whole genome of Streptomyces Ⅱ-2-2-2

[0053] (1) Streptomyces II-2-2-2 was inoculated into sterilized Bennett's liquid medium, and cultured on a shaker at 28° C. and 220 rpm for 48 hours.

[0054] (2) Centrifuge the bacterial solution, wash the precipitate with PBS buffer for 2-3 times, transfer the precipitate to a 1.5mL sterile cryopreservation tube until the bacterial precipitate reaches a height of 0.5mL, and enrich to obtain Streptomyces Ⅱ-2- The whole genome of Streptomyces Ⅱ-2-2-2 was detected, and some genes were annotated.

[0055] Genome-wide detection results: The final assembled genome of Streptomyces II-2-2-2 consists of a 4409970bp linear DNA chromosome with a G+C content of 66.43%, including 3974 predicted coding genes, 72 tRNA and 10 rRNA genes . cgview of the whole genome of Streptomyces II-2-2-2 as Figure 5 shown.

Embodiment 3

[0056] Example 3: Annotation of Streptomyces II-2-2-2 secondary metabolite synthesis gene cluster

[0057] The complete genome sequence of Streptomyces II-2-2-2 was annotated with gene clusters involved in secondary metabolite biosynthesis using antiSMASH 6.0.2. AntiSMASH functional analysis found that Streptomyces II-2-2-2 had five gene clusters involved in the synthesis of secondary metabolites, including LAP, RiPP-like, NRPS, Lanthinipeptides and Arylpolyene. Annotation of Streptomyces II-2-2 secondary metabolite biosynthesis gene cluster as Figure 6 shown.

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Abstract

The invention discloses a new streptomycete strain as well as a separation method and application thereof, the new streptomycete strain is named as streptomycete II-2-2-2 and preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.23479, and the preservation date is September 24, 2021. The streptomyces II-2-2-2 is yellowish brown, small and round on a Gause No.1 solid culture medium, the surface is wrinkled and free of gloss, the middle is raised, filamentous cells exist on the edge, and the streptomyces has milk-color aerial hyphae and dark brown matrix hyphae, and is relatively compact in texture, dry in surface and relatively firm. Under microscopic observation, the diameter of the mycelium is 0.7-0.9 mu m, the diameter of the spore is 0.4-0.6 mu m, and gram staining shows positive. Experiments prove that the streptomyces sp. II-2-2-2 has rich natural product production capacity and can be used for antibiotic mining, and secondary metabolites and biosynthetic gene clusters are worthy of further research and analysis.

Description

technical field [0001] The invention belongs to the field of microbial pharmacy, and in particular relates to a new streptomyces strain and its isolation method and application. Background technique [0002] Actinomycetes are the largest, most numerous, and most widely distributed bacteria among known organisms. So far, the lifestyle, morphology, physiological and metabolic activities and genetic analysis of actinomycetes have become research hotspots. Research on new molecular biology techniques and methods based on actinomycete genomes is the frontier of biomedical science and technology in the 21st century. Since the 1950s, thanks to the extensive research and screening of secondary metabolites of microorganisms in common terrestrial habitats, many important antibiotics, anti-tumor and immunomodulatory drugs have been discovered and applied clinically, helping humans overcome stubborn and serious diseases. Disease provides the material basis. However, due to the widesp...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02G01N30/02G01N30/06G01N30/30G01N30/32G01N30/34G01N30/72A61K35/741A61P31/04C12R1/465
Inventor 江志波胡运琪吴秀丽贾荣亮马晓莉南泽东李舂龙郭欢欢
Owner BEIFANG UNIV OF NATITIES