Morchellaconica extracellular polysaccharide extractive and preparation method and application thereof
A technology of Morchella splendens and extracellular polysaccharide, applied in the biological field, can solve problems such as unfavorable separation and purification and identification of physiological efficacy, large influence of polysaccharide composition and content, difficulty in artificial cultivation, etc., and achieves easy separation and purification, accurate composition, Conditionally controllable effects
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Embodiment 1
[0033] A preparation method for the extracellular polysaccharide extract of Morchella apex, the steps are as follows:
[0034] (1) Morchella conica GIM5.70 (Morchella conica) (purchased from Guangdong Microbial Culture Collection Center.) Fermentation liquid was obtained through slant or plate culture, liquid shaking culture and liquid fermentation culture.
[0035] The specific medium formula and culture conditions are as follows:
[0036] ①Slant or plate culture: medium formula: 200g potato, 20g glucose, 1g peptone, (NH 4 ) 2 SO 42g, MgSO 4 ·7H 2 O 1g, KH 2 PO 4 1g, 20g of agar, add distilled water to make the volume to 1000ml, and adjust the pH to 6.5; then insert a piece of morel mycelia with a diameter of about 4mm, culture at 26°C for 14 days, and store at 4°C for later use.
[0037] ②Liquid shaking culture: with NH 4 NO 3 As a nitrogen source, glucose or soluble starch as a carbon source, and initial pH and inoculum size as investigation factors, the final form...
Embodiment 2
[0047] A preparation method for the extracellular polysaccharide extract of Morchella apex, the steps are as follows:
[0048] (1) The Morchella cuspidatum is cultured on a slant or flat plate, liquid shaking culture and liquid fermentation culture. Concrete medium formula and culture condition are the same as embodiment 1.
[0049] (2) separating the mycelium from the fermented product through centrifugation and vacuum filtration to obtain a clarified fermented liquid. Concrete method is with embodiment 1.
[0050] (3) Concentrating the fermented broth: use a rotary evaporator to concentrate under reduced pressure in a constant temperature water bath at 50°C to 1 / 3 of the original volume.
[0051] (4) protein removal, centrifugation: with embodiment 1.
[0052] (5) Add 1 volume of 95% ethanol to the active extract after deproteinization, dialyze and settle at 0°C for 15 hours, the pH value is 4, let it stand overnight, and precipitate a flocculent white substance, which is...
Embodiment 3
[0055] A preparation method for the extracellular polysaccharide extract of Morchella apex, the steps are as follows:
[0056] (1) The Morchella cuspidatum is cultured on a slant or flat plate, liquid shaking culture and liquid fermentation culture. Concrete medium formula and culture condition are the same as embodiment 1.
[0057] (2) separating the mycelium from the fermented product through centrifugation and vacuum filtration to obtain a clarified fermented liquid. Concrete method is with embodiment 1.
[0058] (3) Concentrating the fermented broth: use a rotary evaporator to concentrate under reduced pressure in a constant temperature water bath at 70°C to 1 / 6 of the original volume.
[0059] (4) protein removal, centrifugation: with embodiment 1.
[0060] (5) Add 5 times the volume of 95% ethanol to the active extract after deproteinization, dialyze and settle at 10°C for 25 hours, the pH value is 8, let it stand overnight, and precipitate a flocculent white substanc...
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