A fungus dym25 with antibacterial, antioxidant and anticancer effects and its application
A DYM25, anti-oxidation technology, applied in the direction of application, fungi, biochemical equipment and methods, etc., can solve the problem of resistance to cucumber wilt that has not been reported
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Embodiment 1
[0029] Example 1 Screening and identification of Talaromyces sp.DYM25
[0030] The Talaromyces sp.DYM25 of the present application was screened and isolated from the seawater samples of the Yap Trench in the central and western Pacific Ocean during the 150th dive of Jiaolong. Combined with the analysis of molecular biological information of morphology and ITS gene sequence, the strain was finally identified as basket-shaped Talaromyces sp..
[0031] 1. Isolation and purification of strains: Take 0.5 mL of seawater samples (concentrated) at a depth of 6,000 meters in the Yapu Trench, and perform gradient dilution with sterile seawater by the ten-fold dilution method. Take 100 μL of the diluted samples and spread them on the fungal separation and culture. The fungi were isolated and purified according to the different colony morphology, and were cultured upside down at 28°C for 5-7 days.
[0032] Wherein, the fungal isolation medium is PDA medium (BD Difco brand). Sterile seaw...
Embodiment 2
[0045] Example 2 Stability of extracellular active substances of Talaromyces sp.DYM25
[0046] Strain DYM25 was inoculated in PDB medium, 28 ℃, 180r / min shaker culture for 3 days, the cultured bacterial liquid was inoculated in PDB medium at 10% (v:v) inoculum, 28 ℃, 180r / min fermentation After culturing for 7 days, the fermentation broth was passed through a 200-mesh sieve to obtain a clarified fermentation supernatant.
[0047] 1. The effect of temperature on the active substances: Take 4 parts of the clarified fermentation supernatant of strain DYM25, respectively, after treatment at 60°C, 80°C, 100°C, and 120°C for 30 minutes, filter them through 0.22 μm at a ratio of 1:20 (v:v). The film was added to the PDA medium cooled to about 50°C, poured into the plate, and the antibacterial activity was measured according to the mycelial growth rate method.
[0048] The result is as Figure 4 As shown, the bacteriostatic activity of the active substances in the fermentation super...
Embodiment 3
[0051] Example 3 Preparation of crude extract of Talaromyces sp.DYM25
[0052] 1. Activation of Talaromyces sp. DYM25: The strain DYM25 was inoculated in PDB medium and cultured for 3 days at 28°C and 180r / min on a shaker.
[0053] 2. Preparation of fermentation supernatant: the bacterial liquid in step 1 was inoculated into PDB medium according to the inoculum amount of 10% (v:v), fermented and cultured at 28° C., 180r / min for 7d, and the fermentation broth passed through a 200-mesh sieve. The supernatant was obtained by removing the bacterial cells, which was filtered through a Buchner funnel to obtain a clarified fermentation supernatant.
[0054] 3. Preparation of crude extract: in the clarified fermentation supernatant obtained in step 2, add an equal volume of water-saturated n-butanol solution for extraction, and invert repeatedly 3 to 4 times and then stand for stratification to obtain an organic phase (extraction). Phase) and the water phase (raffinate phase), the wa...
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