Fungus culture extract and preparation method and application thereof
A fungal culture and extract technology, applied in the field of biopharmaceuticals, can solve problems such as large side effects
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Embodiment 1
[0055] Example 1: Preparation and structural identification of extracts from fungal cultures
[0056] 1. Preparation of extracts from fungal cultures
[0057] 1. Seed culture:
[0058] (1) Preparation of seed medium: 2.1% glucose, 1.9% peptone, 0.6% yeast extract, 2.5% sea salt, sterilized by autoclaving at 121°C for 20 minutes to obtain sterilized seed medium;
[0059] (2) Seed cultivation: After the cryopreserved deep-sea fungus CCTCC M 2015628 was resuscitated, the strain was inserted into the above-mentioned seed medium, and cultured on a shaking table at 29°C and 220r / min for 49 hours to obtain a seed culture solution.
[0060] 2. Fermentation culture:
[0061] (1) Preparation of fermentation medium (liquid medium): weigh sea salt, glucose, peptone, and yeast extract and dissolve them in deionized water. After complete dissolution, adjust the pH to 7.6, and sterilize under high pressure at 121°C for 30 minutes. Inoculate according to the ratio of every 5mL seed culture...
Embodiment 2
[0078] Embodiment 2: Experiment on the anti-inflammatory activity of the cyclic dipeptide compounds obtained in Example 1
[0079] In this embodiment, the anti-inflammatory activity detection includes three parts:
[0080] (1) Measurement of nitrite standard curve; (2) Detection of NO production; (3) MTT method to detect the toxicity of samples to cells.
[0081] (1) Measurement of nitrite standard curve:
[0082] Dilute the 0.1M nitrite solution to 100μM with buffer, and take 1ml of the 100μM nitrite solution. Select 3 rows of wells on the 96-well plate, add 50 μl of buffer solution to the wells of B-H respectively, and add 50 μl of 100 μM nitrite solution to the wells of row A respectively. According to the order of A-H, dilute the nitrite 2 times, that is, take 50 μl of the solution from the previous row of wells and add it to the next row of wells. The final concentration of nitrite in the holes of A-H is 100, 50, 25, 12.5, 6.25, 3.13 and 1.56 μM, the solution volume in...
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