A saffron endophytic fungus and its crude polysaccharide, preparation method and application
An endophytic fungus, saffron technology, applied in biochemical equipment and methods, microorganism-based methods, fungi, etc., to achieve the effects of scavenging free radicals, inhibiting tumor cell proliferation, and reducing product costs
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Embodiment 1
[0040] Embodiment 1: the preparation of the crude polysaccharide of saffron endophytic fungus
[0041] 1. Preparation of medium:
[0042] PDA culture medium and PYG liquid culture medium described in the present invention are conventional culture medium, can be prepared by following method.
[0043] (1) PDA medium (potato dextrose agar): 200g of potatoes (peeled, cut into small pieces, boiled in deionized water for 20min, filtered through gauze, discarded the mashed potatoes, and took the filtrate), 10g of glucose, 15g of agar, fixed in deionized water Capacity to 1000mL.
[0044] (2) PYG liquid medium: yeast extract 1g, peptone 2g, glucose 10g, sodium chloride 2g, pH 6.8-7.2, dilute to 1000mL with deionized water.
[0045] All the above mediums were sterilized at a temperature of 121° C. and a pressure of 0.1 MPa for 20 minutes.
[0046] 2. Activation of strains:
[0047] The endophytic fungus is isolated from healthy saffron bulbs through a strict surface disinfection meth...
Embodiment 2
[0058] Embodiment 2: the activity of endophytic fungus crude polysaccharide of saffron
[0059] This embodiment is a series of in vitro antioxidant tests carried out on the saffron endophytic fungus crude polysaccharide prepared in Example 1, and the obtained test results are as follows Figure 2 ~ Figure 6 shown.
[0060] 1. Test method
[0061] (1) Scavenging test for DPPH free radicals
[0062] Add 0.5mL of crude polysaccharide solutions of different concentrations and 3.5mL of 0.1mmol / L DPPH solution dissolved in 80v / v% ethanol to the sample tube, the final concentrations of crude polysaccharide are 0.025, 0.05, 0.1, 0.2, 0.3, 0.4 and 0.5 mg / mL; 80v / v% ethanol in the control group; 0.5mL distilled water was used to replace the crude polysaccharide solution and 3.5mL 0.1mmol / L DPPH solution dissolved in 80v / v% ethanol in the blank group. After mixing the solutions of each of the above experimental groups, put them in a dark place at room temperature for 30 minutes. Adju...
Embodiment 3
[0081] Embodiment 3: Antibacterial test in vitro of the crude polysaccharide of saffron endophytic fungus
[0082] 1. Test method
[0083] (1) Test strains: Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Escherichia coli aerogenes, Salmonella typhi, Shigella, Klebsiella pneumoniae (purchased from China Institute for the Control of Pharmaceutical and Biological Products , and the rest were purchased from Wuhan Culture Collection, China).
[0084] (2) Preparation of bacteria suspension: Activation and culture of test strains were carried out for 24 hours with LB plate medium at 37°C. The activated bacteria were mixed with sterile saline to obtain a certain concentration of bacteria suspension (about 0.5 McFarland standard solution).
[0085] (3) Preparation of paper sheets containing samples: ordinary qualitative filter paper was punched into a number of circular paper sheets with a diameter of 6.0 mm, used as blank paper sheets, sterilized...
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