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A kind of extract of fungal culture and its preparation method and application
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A fungal culture and extract technology, applied in the field of biopharmaceuticals, can solve problems such as large side effects
Active Publication Date: 2020-02-07
SHENZHEN UNIV
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However, most of these inhibitors are limited to chemically synthesized substances with relatively 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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Abstract
The present invention is applicable to the field of biopharmaceuticals, and provides an extract of a fungal culture and its preparation method and application. The extract is a cyclic dipeptide compound, and the cyclic dipeptide compound can be numbered as CCTCC M 2015628 It is isolated from deep-sea fungal fermentation culture. The cyclic dipeptide compound has strong activity of inhibiting the production of nitric oxide (NO), and is non-toxic to cells; it can be used for the preparation of drugs for treating Alzheimer's disease.
Description
technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to an extract of fungal culture and its preparation method and application. Background technique [0002] Daily inflammatory diseases that currently plague people include joint inflammation such as osteoarthritis, rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis, etc.; inflammatory skin diseases such as eczema, psoriasis, dermatitis, etc.; inflammatory eye diseases such as Uveitis, conjunctivitis, etc.; lung diseases such as asthma, bronchitis, acute respiratory distress syndrome, etc.; bacteremia, that toxemia, aphthous ulcers, gingivitis, pancreatitis, etc.; Ileitis, atrophic gastritis, ulcerative colitis, celiac disease, peptic ulcer, irritable bowel syndrome such as mucosal inflammation caused by Helicobacter pylori infection or gastrointestinal tract caused by non-steroidal anti-inflammatory drugs illness and so on. In recent years, studies on neurodegenerative ...
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