Streptomyces lydicus producing natamycin and uses thereof
A technology of streptomyces lidi and natamycin, applied in the field of microorganisms, can solve the problems of ineffectiveness of bacteria and viruses
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Embodiment 1
[0037] Embodiment 1, separation and preparation of Streptomyces lydicus (Streptomyces lydicus) A01 CGMCC No.1653
[0038] Collect soil samples from vegetable fields in Miyun County, Beijing, take 10g and add them to a conical flask containing 100ml sterile water and an appropriate amount of glass beads, place on a shaker and vibrate at 100rpm for 20min, take 0.5ml and add them to 4.5ml sterile water, and then dilute for 10 2 、10 3 、10 4 Take 0.1ml of the above soil suspension and spread it evenly on the Gaoshi No. 1 medium plate, and blow it in the ultra-clean workbench until it is slightly dry; each concentration is repeated 3 times, and placed in a 28°C incubator for 5- On the 10th day, a single colony of actinomycetes was picked for dilution, streaking, isolation and purification to obtain a pure culture strain.
[0039] Preliminary screening: The obtained pure culture strains were subjected to shake flask fermentation culture, and the fermentation liquid was filtered and...
Embodiment 2
[0057] Embodiment 2, the bacteriostatic activity assay of Streptomyces lydicus (Streptomyces lydicus) A01 CGMCC No.1653 fermented liquid
[0058] Slant culture: Streptomyces lydicus (Streptomyces lydicus) A01 CGMCC No.1653 was inoculated on Gao's No. 1 slant medium, and cultured at 28°C for 7-10 days until it produced a sufficient amount of spores;
[0059] Seed culture: Scrape 3-5 rings of spores with a sterile platinum ring and inoculate them into a 500ml Erlenmeyer flask containing 100ml of seed medium, place on a temperature-controlled shaker, and at 28°C, with a rotation radius of 13mm and 180rpm Shake culture at constant temperature at a constant speed for 20-24 hours;
[0060] Fermentation culture: Take the above-mentioned seed culture solution and inoculate it into a 500ml Erlenmeyer flask containing 100ml of fermentation medium at an inoculum size of 3% (V / V). Under the condition of 28°C, shake at a constant temperature with a rotation radius of 13mm and a speed of 18...
Embodiment 3
[0062] Embodiment 3, the antibacterial mechanism of Streptomyces lydicus (Streptomyces lydicus) A01 CGMCC No.1653 metabolites
[0063] 1. Crude extraction and preliminary identification of natamycin in fermentation broth
[0064] The fermented broth obtained in Example 2 was pretreated with 3 times the volume of absolute ethanol, and stood at 4°C for 2 hours to precipitate bacteria, solid particles, soluble viscose, nucleic acids, miscellaneous proteins, and intermediate metabolites, etc., above The supernatant was vacuum-filtered with a Buchner funnel with two layers of filter paper, and the filtrate was concentrated under reduced pressure at 45°C by a rotary evaporator.
[0065] The chemical type of the active substance was identified by paper chromatography of the Czech eight-solvent system, and the ultraviolet full-wavelength scanning was carried out at 190nm to 400nm.
[0066] 1. Paper chromatography of the Czech eight-solvent system
[0067] Solvent system is: I. water...
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