A kind of Micromonospora tmd166 and its application
A technology of TMD166, Micromonospora, applied in the fields of microbiology and pharmacy, to achieve the effect of strong inhibitory activity and good development prospects
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Embodiment 1
[0048] The isolation of embodiment 1 Micromonospora bacterial strain TMD166
[0049] The camel thorns collected from the Taklamakan Desert were washed with water, and the surface impurities were removed by ultrasonic waves. The samples were soaked in 1% Tween-20 for 1 minute, sodium hypochlorite aqueous solution containing 0.4% available chlorine for 7 minutes, and 2.5% sodium thiosulfate. Soak for 9 minutes, soak in 75% ethanol for 2 minutes, soak in 10% sodium bicarbonate for 10 minutes, wash with sterile water 3 times after each soak. After the sample was dried, it was ground into powder with a pulverizer and evenly sprinkled on the M-WA medium (glycerol 10g, yeast 0.5g, proline 1.0g, aspartic acid 1.0g, potassium nitrate 0.5g, pyruvate Sodium 1.25g, betaine 1.25g, agar 20g, sterile water 1L, pH 8.0, sterilized at 121°C for 30 minutes), incubated at 28°C. At the same time, after surface disinfection, 200 μl of distilled water for the third wash was taken, spread on the ISP...
Embodiment 2
[0050] Identification of Example 2 Micromonospora bacterial strain TMD166
[0051] (1) Morphological identification:
[0052] Micromonospora TMD166 of the present invention, such as figure 1 shown. The colony on the ISP-2 medium shows a light orange color, and the surface is covered with a layer of powdery spores, and grows rapidly at a culture temperature of 28-30°C.
[0053] (2) Molecular identification:
[0054] The genomic DNA was extracted, amplified and analyzed using universal primers 27f and 1492r, and a single DNA band was obtained. In the GenBank database, carry out Blast comparison, screen the typical strain sequence with the highest similarity, then use MEGA5.2 software to carry out cluster analysis and construct phylogenetic tree with neighbor method (Neighbor-joining), such as figure 2 As shown, TMD166 is associated with Micromonospora polyrhachis NEAU-ycm2 T Gathered in the same branch, with a similarity of 99.48%, the endophyte TMD166 was finally identifi...
Embodiment 3
[0055] The fermentation of embodiment 3 Micromonospora bacterial strain TMD166
[0056] Micromonospora strain TMD166 grown in the slant medium was inoculated on the seed medium, and the seed medium was No. 38 medium (glucose 4.0g, yeast powder 4.0g, malt extract powder 5.0g, ZnSO 4 .7H 2 O 0.001g, MnCl 2 .4H 2 O0.001g, FeSO 4 .7H 2 O 0.002g, phenylalanine 0.001g, alanine 0.0003g, vitamin B1 0.001g, vitamin B20.001g, vitamin B6 0.001g, niacin 0.001g, biotin 0.001g, 1.0L sterile distilled water, pH 8.5 ). Put 100mL No. 38 medium in each 500mL shake flask, culture on a 28-30°C, 180rpm rotary shaker for 72h, and then transfer to the fermentation medium (same as the seed medium) with 10% inoculum amount, and put in each 5.0L shake flask Pack 1.0L of fermentation medium, culture at 28-30°C on a rotary shaker at 180rpm for 96h, and harvest the fermentation broth.
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