Morchellaconica extracellular polysaccharide extractive and preparation method and application thereof

Abstract

The invention belongs to the biological technical field, which discloses a morchellaconica extracellular polysaccharide extractive and a preparation method and application thereof. The extractive is liquid fermentation liquor which is obtained by fermenting and culturing morchellaconica liquid and separating mycelium through centrifugation and vacuum filtration. The liquid fermentation liquor is concentrated to 1 / 3 to 1 / 6 of the original bulk to obtain concentrated solution and the concentration temperature is 50 to 70 degrees centigrade. The protein of the concentrated solution is removed and the concentrated solution is centrifuged. Activated extract is obtained. Polysaccharide in the concentrated solution is dialyzed and settled in 95 percent of ethanol at 0 to 10 degrees centigrade for 12 to 25 hours, the bulk of which is once to 5 times larger than the bulk of the activated extract. The pH value is 4 to 8. Flocculent white substance is precipitated after still placement. White sedimentation is obtained after centrifugation separation, which is frozen and dried to obtain the morchellaconica extracellular polysaccharide extractive. The invention has simple preparation process and allows mass production. The extractive has no harm on human body and good antioxidant and anti-aging action and can be used for preparing antioxidant and anti-aging medicine or health care food.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a fungal extract with medicinal value, in particular to a preparation method of an exopolysaccharide extract from Morchella conica and its application in anti-oxidation and anti-aging. Background technique

[0002] For the extraction and biological activity of edible fungus polysaccharides, most of the existing reports are to extract polysaccharides from the fruiting bodies of edible fungi. The tea tree mushroom polysaccharide obtained by ethanol precipitation (Journal of Edible Fungi, 2006, 13 (4): 63-68); another example is the morel mycelium and fermentation liquid produced by submerged fermentation of semi-synthetic medium by Jia Jianhui et al. Morel polysaccharides were extracted (Food Science, 2002, 23(4): 59-62). On the one hand, the use of polysaccharides obtained from fruiting bodies consumes a lot of resources. At present, the fruiting bodies of Morchella apexus are diffic...

Examples

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...

technical field

[0001] The invention relates to the field of biotechnology, in particular to a fungal extract with medicinal value, in particular to a preparation method of an exopolysaccharide extract from Morchella conica and its application in anti-oxidation and anti-aging. Background technique

[0002] For the extraction and biological activity of edible fungus polysaccharides, most of the existing reports are to extract polysaccharides from the fruiting bodies of edible fungi. The tea tree mushroom polysaccharide obtained by ethanol precipitation (Journal of Edible Fungi, 2006, 13 (4): 63-68); another example is the morel mycelium and fermentation liquid produced by submerged fermentation of semi-synthetic medium by Jia Jianhui et al. Morel polysaccharides were extracted (Food Science, 2002, 23(4): 59-62). On the one hand, the use of polysaccharides obtained from fruiting bodies consumes a lot of resources. At present, the fruiting bodies of Morchella apexus are diffic...

Examples

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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