A method for improving the content of crude polysaccharide in cordyceps militaris fruiting body residues

Abstract

The present application belongs to the technical field of deep processing of Cordyceps militaris, and provides a method for improving the content of crude polysaccharide in Cordyceps militaris fruiting body residues. The method comprises the steps of cutting, enzyme extraction, acid adjustment, drying, crushing and irradiation. The method opens the spatial structure of dietary fibers such as hemicellulose, cellulose and lignin through enzyme extraction process, exposes the polar groups, and then combines acid treatment, crushing and low-dose irradiation. Under the synergistic effect of various parameters, high-content crude polysaccharide Cordyceps militaris fruiting body residue raw materials are finally obtained. The Cordyceps militaris fruiting body residues produced by the method greatly improve the content of crude polysaccharide active ingredients, thereby improving the added value of Cordyceps militaris production.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of Cordyceps militaris deep processing, and particularly relates to a method for improving the content of crude polysaccharide in Cordyceps militaris fruiting body residues. BACKGROUND

[0002] The Cordyceps militaris fruiting body residue is a by-product produced by a Cordyceps militaris deep processing enterprise, that is, after Cordyceps militaris fruiting bodies are harvested and dried, the Cordyceps militaris fruiting bodies are processed through crushing, extraction, filtration, water squeezing and drying, and the like, and the main components are carbohydrates, proteins and fats, and the like. After the Cordyceps militaris fruiting bodies are extracted, the remaining residue contains a relatively high content of proteins and dietary fibers (cellulose, hemicellulose and lignin). At the present stage, the Cordyceps militaris fruiting body residue is mostly discarded, and a small amount is used for animal feed, and the utilization efficiency and practical value are particularly low.

[0003] As a sterilization technology in the field of food, the irradiation technology is to make the electron beam generated by an electron accelerator (0.2-10 MeV) act on the irradiated food raw materials or products, so that the irradiated substances undergo a series of physical, chemical and biochemical changes, resulting in degradation, polymerization, cross-linking and modification of the substances. In terms of the irradiation technology for modifying polysaccharides, the effect of irradiation is mainly affected by factors such as raw materials, irradiation dose, temperature, oxygen content, moisture content and pH. Among them, the raw materials, the irradiation dose and the pH have a greater influence on the degradation and solubilization of polysaccharides.

[0004] At present, there are relatively few reports on the treatment of Cordyceps militaris mycelium or fruiting bodies by using the irradiation technology. 60 Effect of Co-γ irradiation on four bioactive components in Cordyceps militaris fruiting bodies, shows that the dose of 0-20 kGy has no significant effect on the content of polysaccharides, cordycepic acid, cordycepin and adenosine in the dried Cordyceps militaris fruiting bodies (Wang P, Liu GJ, Zhou SJ, Qiao YC, Yang SL. 60 Effect of Co-γ irradiation on four bioactive components in Cordyceps militaris fruiting bodies, shows that the dose of 0-20 kGy has no significant effect on the content of polysaccharides, cordycepic acid, cordycepin and adenosine in the dried Cordyceps militaris fruiting bodies (Wang P, Liu GJ, Zhou SJ, Qiao YC, Yang SL. 60Co-γ ray irradiation on the biological active ingredients of Cordyceps militaris fruiting bodies [J]. Anhui Agricultural Science, 2021, 49(20): 202-204.) Hsiang-Yu Lin et al. reported that the Cordyceps militaris mycelium was treated by irradiation technology, and when the irradiation dose increased to 20 kGy, the polysaccharide content of Cordyceps militaris mycelium increased by 50.9%, and the molecular weight distribution was expanded (Lin H Y, Tsai S Y, Tseng Y L, et al. Gamma irradiation for improving functional ingredients and determining the heat treatment conditions of Cordyceps militaris mycelia [J]. Journal of Thermal Analysis and Calorimetry, 2015, 120(1): 439-448.). Yin Zhaomin et al. provided a preparation method of edible mushroom polysaccharide with high purity, high solubility and high activity (the raw materials include Cordyceps militaris fruiting bodies, mycelium, stipe and culture medium), which is treated by γ ray (Co) and combined with biological enzyme complex enzymolysis, ultra-high speed shearing and other technologies, so that the dissolution rate of edible mushroom polysaccharide is increased by more than 20%, thereby the product yield is greatly improved (Yin Zhaomin, Gao Hong, Yao Fen, et al. A preparation method of edible mushroom polysaccharide with high purity, high solubility and high activity: CN109694416A [P]. 2019.). But the defects of the above-mentioned technology are high required irradiation dose and high application cost. 60 Co) treatment, and combined with biological enzyme complex enzymolysis, ultra-high speed shearing and other technologies, so that the dissolution rate of edible mushroom polysaccharide is increased by more than 20%, thereby the product yield is greatly improved (Yin Zhaomin, Gao Hong, Yao Fen, et al. A preparation method of edible mushroom polysaccharide with high purity, high solubility and high activity: CN109694416A [P]. 2019.). But the defects of the above-mentioned technology are high required irradiation dose and high application cost. SUMMARY

[0005] In order to overcome the above-mentioned defects in the prior art, the present application provides a method for improving the content of crude polysaccharide in Cordyceps militaris fruiting body residues.

[0006] In order to achieve the above-mentioned application purposes, the present application provides the following technical solutions:

[0007] The present application provides a method for improving the content of crude polysaccharide in Cordyceps militaris fruiting body residues, comprising the following steps:

[0008] (1) cutting fresh Cordyceps militaris fruiting bodies to obtain Cordyceps militaris fruiting body segments;

[0009] (2) mixing the Cordyceps militaris fruiting body segments with water, and then mixing with extraction enzymes for enzyme extraction to obtain an enzyme extraction liquid;

[0010] (3) adjusting the pH value of the enzyme extraction liquid to 3-5, and filtering to obtain Cordyceps militaris fruiting body residues;

[0011] (4) sequentially drying and crushing the Cordyceps militaris fruiting body residue to obtain Cordyceps militaris fruiting body residue powder;

[0012] (5) performing irradiation treatment on the Cordyceps militaris fruiting body residue powder.

[0013] Preferably, the length of the Cordyceps militaris fruiting body segment in step (1) is 1-5 cm.

[0014] Preferably, the mixing mass ratio of the Cordyceps militaris fruiting body segment to water in step (2) is 1:3-5.

[0015] Preferably, the extraction enzyme in step (2) comprises one or more of beta-glucanase, xylanase, pectinase, cellulase and hemicellulase, and the amount of the extraction enzyme is 0.01-0.5% of the dry weight of the Cordyceps militaris fruiting body segment.

[0016] Preferably, the temperature of the enzyme extraction in step (2) is 50-55℃, and the enzyme extraction time is 2-4 h.

[0017] Preferably, the component for adjusting the pH value in step (3) is edible acid, and the edible acid comprises one or more of citric acid, malic acid, gluconic acid, lactic acid, tartaric acid, phosphoric acid, fumaric acid, adipic acid and succinic acid.

[0018] Preferably, the drying temperature in step (4) is 45-65℃.

[0019] Preferably, the moisture of the residue after drying in step (4) is 10-15%.

[0020] Preferably, the crushing in step (4) is first coarse crushing and then ultrafine crushing, the mesh number of the coarse crushing is 60-80 mesh, and the mesh number of the ultrafine crushing is 150-500 mesh.

[0021] Preferably, the irradiation treatment in step (5) is gamma ray treatment using a Co irradiation device, and the irradiation dose during the irradiation treatment is 4-18 kGy. 60 Co irradiation device, and the irradiation dose during the irradiation treatment is 4-18 kGy.

[0022] Compared with the prior art, the present application has the following beneficial effects:

[0023] (1) The method of the present application opens the spatial structure of dietary fibers such as hemicellulose, cellulose and lignin through the enzyme extraction process, exposes the polar groups, and then combines acid treatment, crushing and low-dose irradiation, so that, under the synergistic effect of various parameters, a high-content crude polysaccharide Cordyceps militaris fruiting body residue raw material is finally obtained. The Cordyceps militaris fruiting body residue produced by the method of the present application greatly improves the effective components of crude polysaccharides, thereby increasing the added value of Cordyceps militaris production.

[0024] (2) Adopt low irradiation dose, avoid Cordyceps militaris fruiting body residue due to irradiation dose too large, resulting in protein denaturation, thereby producing "smell".

[0025] (3) Compared with culture medium and cultivation process optimization to improve polysaccharide, the method is simple in operation, and the polysaccharide content is more stable. DETAILED DESCRIPTION

[0026] The application provides a method for improving the content of crude polysaccharide of Cordyceps militaris fruiting body residue, comprising the following steps:

[0027] (1) cutting fresh Cordyceps militaris fruiting body to obtain Cordyceps militaris fruiting body segments;

[0028] (2) mixing the Cordyceps militaris fruiting body segments with water, and then mixing with an extraction enzyme to perform enzyme extraction to obtain an enzyme extraction liquid;

[0029] (3) adjusting the pH value of the enzyme extraction liquid to 3-5, and filtering to obtain Cordyceps militaris fruiting body residue;

[0030] (4) sequentially drying and crushing the Cordyceps militaris fruiting body residue to obtain Cordyceps militaris fruiting body residue powder;

[0031] (5) performing irradiation treatment on the Cordyceps militaris fruiting body residue powder.

[0032] In the application, the length of the Cordyceps militaris fruiting body segments in step (1) is preferably 1-5 cm, further preferably 2-4 cm, and more preferably 3 cm.

[0033] In the application, the mixing mass ratio of the Cordyceps militaris fruiting body segments to water in step (2) is preferably 1:3-5, and further preferably 1:4.

[0034] In the application, the extraction enzyme in step (2) preferably comprises one or more of beta-glucanase, xylanase, pectinase, cellulase and hemicellulase, and is further preferably beta-glucanase; the dosage of the extraction enzyme is preferably 0.01-0.5% of the dry weight of the Cordyceps militaris fruiting body segments, further preferably 0.06-0.2% of the dry weight of the Cordyceps militaris fruiting body segments, and more preferably 0.1% of the dry weight of the Cordyceps militaris fruiting body segments.

[0035] In the application, the temperature of the enzyme extraction in step (2) is preferably 50-55 DEG C, further preferably 52-54 DEG C, and more preferably 53 DEG C; and the time of the enzyme extraction is preferably 2-4 h, further preferably 2.5-3.5 h, and more preferably 3 h.

[0036] In this invention, step (3) involves adjusting the pH of the enzyme extract to 3-5, filtering, and obtaining Cordyceps militaris fruiting body residue. More preferably, the enzyme extract is adjusted to pH 4, filtered, and obtained Cordyceps militaris fruiting body residue.

[0037] In this invention, the component for adjusting the pH value in step (3) is preferably an edible acid, which preferably includes one or more of citric acid, malic acid, gluconic acid, lactic acid, tartaric acid, phosphoric acid, fumaric acid, adipic acid and succinic acid, and is more preferably malic acid.

[0038] In this invention, the drying temperature in step (4) is preferably 45-65°C, more preferably 50-60°C, and even more preferably 55°C.

[0039] In this invention, the drying in step (4) is preferably to a residual moisture content of 10-15%, more preferably to a residual moisture content of 12-14%, and even more preferably to a residual moisture content of 13%.

[0040] In this invention, the pulverization in step (4) is preferably performed by first coarse pulverization and then by ultrafine pulverization. The mesh size of the coarse pulverization is preferably 60-80 mesh, more preferably 65-75 mesh, and even more preferably 70 mesh; the mesh size of the ultrafine pulverization is preferably 150-500 mesh, more preferably 325-480 mesh, and even more preferably 450 mesh.

[0041] In this invention, the irradiation treatment in step (5) is preferably performed using... 60 The γ-ray treatment is performed using a Co irradiation device, wherein the irradiation dose during the irradiation process is preferably 4–18 kGy, more preferably 5–10 kGy, and even more preferably 6 kGy.

[0042] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0043] The Cordyceps militaris fruiting bodies in the following examples and comparative examples were produced by Tianshui Zhong'an Biotechnology Co., Ltd., and the parent strain was sourced from Changde Yandi Biotechnology Co., Ltd.

[0044] Example 1

[0045] (1) Cutting: After the fresh Cordyceps militaris fruiting bodies are harvested, they are cut into 1cm segments using a shredder.

[0046] (2) Enzymatic extraction: after the segmented Cordyceps militaris fruiting bodies were added 4 times Cordyceps militaris fruiting body mass of water, and then added 0.1% of the dry weight of the Cordyceps militaris fruiting body segments β-glucanase, pH natural, extraction temperature 53℃, extraction time 3h.

[0047] (3) Acid adjustment: after the extraction was completed, the pH of the extract was adjusted to 4 using malic acid, the filtrate was removed, and the Cordyceps militaris fruiting body residue was dried at 55℃ until the moisture content of the residue was 13%.

[0048] (4) Grinding: the Cordyceps militaris fruiting body residue was first coarsely ground to obtain 70 mesh coarse powder, and then the coarse powder was further ground using an air jet mill, with 90% of the ground particles passing through a 450 mesh.

[0049] (5) Packaging: the fine powder passing through the 450 mesh was independently packaged, with a loading capacity of 300g per box, a fine powder thickness of 8cm, and 10 air vents on the packaging box to ensure sufficient oxygen for the irradiation modification process during irradiation.

[0050] (6) Irradiation: a Co irradiation device was used for γ-ray treatment, with an irradiation dose of 10kGy. 60

[0051] (7) The method for detecting polysaccharides in the Cordyceps militaris fruiting body residue powder was NYT 1676-2008 "Determination of Crude Polysaccharide Content in Edible Mushrooms".

[0052] Example 2

[0053] (1) Cutting: after the fresh Cordyceps militaris fruiting bodies were harvested, a shredder was used to cut the fruiting bodies into 3cm segments.

[0054] (2) Enzymatic extraction: after the segmented Cordyceps militaris fruiting bodies were added 4 times Cordyceps militaris fruiting body mass of water, and then added 0.1% of the dry weight of the Cordyceps militaris fruiting body segments β-glucanase, pH natural, extraction temperature 53℃, extraction time 3h.

[0055] (3) Acid adjustment: after the extraction was completed, the pH of the extract was adjusted to 4 using malic acid, the filtrate was removed, and the Cordyceps militaris fruiting body residue was dried at 55℃ until the moisture content of the residue was 13%.

[0056] (4) Grinding: the Cordyceps militaris fruiting body residue was first coarsely ground to obtain 70 mesh coarse powder, and then the coarse powder was further ground using an air jet mill, with 90% of the ground particles passing through a 450 mesh.

[0057] (5) Packaging: the fine powder passing through the 450 mesh was independently packaged, with a loading capacity of 300g per box, a fine powder thickness of 8cm, and 10 air vents on the packaging box to ensure sufficient oxygen for the irradiation modification process during irradiation.

[0058] ​(6) Irradiation: using 60 Co irradiation device for gamma ray treatment, irradiation dose 6kGy.

[0059] (7) The polysaccharide detection method in the Cordyceps militaris fruiting body residue is NYT 1676-2008 "Determination of the content of crude polysaccharide in edible fungi".

[0060] Example 3

[0061] (1) Cutting: after the fresh Cordyceps militaris fruiting body is harvested, the fruiting body is cut into 5cm segments by using a shredder.

[0062] (2) Enzyme extraction: after the segmented Cordyceps militaris fruiting body is added 5 times the mass of Cordyceps militaris fruiting body water, 0.2% xylanase and 0.3% pectinase are added, the pH is natural, the extraction temperature is 55℃, and the extraction time is 4h.

[0063] (3) Acid adjustment: after the extraction is completed, the pH of the extraction liquid is adjusted to 5 by using tartaric acid, the filtrate is removed, and the Cordyceps militaris fruiting body residue is dried at 65℃ until the moisture content is 15%.

[0064] (4) Grinding: the Cordyceps militaris fruiting body residue is first coarsely ground to obtain 80 mesh coarse powder, and then the coarse powder is further ground by using a super micro grinder, and 80% of the ground powder passes through 400 mesh.

[0065] (5) Packaging: the fine powder passing through 400 mesh is independently packaged, the packaging capacity is 500g / box, the thickness of the Cordyceps militaris fruiting body residue powder is 10cm, and the packaging box has 5 air vents.

[0066] (6) Irradiation: using 60 Co irradiation device for gamma ray treatment, irradiation dose 18kGy.

[0067] (7) The polysaccharide detection method in the Cordyceps militaris fruiting body residue is NYT 1676-2008 "Determination of the content of crude polysaccharide in edible fungi".

[0068] Comparative Example 1

[0069] The difference from Example 1 is that the Cordyceps militaris fruiting body residue is obtained by water extraction (without adding extraction enzyme and without acid treatment), and single irradiation treatment, the irradiation dose is 50kGy.

[0070] Comparative Example 2

[0071] The difference from Example 2 is that the Cordyceps militaris fruiting body residue is obtained by water extraction (without adding extraction enzyme and without acid treatment), and single irradiation treatment, the irradiation dose is 50kGy.

[0072] Comparative Example 3

[0073] The difference from Example 3 is that the Cordyceps militaris fruiting body residues are obtained by water extraction (without adding extraction enzymes, without acid treatment), single irradiation treatment, and the irradiation dose is 50 kGy.

[0074] Experimental Example 1

[0075] The crude polysaccharide contents in the Cordyceps militaris fruiting body residue powders obtained in Examples 1-3 and Comparative Examples 1-3 are compared, and the results are shown in Table 1.

[0076] Table 1. Crude polysaccharide contents obtained by different treatments

[0077]

[0078] As can be seen from Table 1, compared with Comparative Examples 1-3, the method of Examples 1-3 of the present application can significantly increase the crude polysaccharide content in the Cordyceps militaris fruiting body residue powder while reducing the irradiation dose.

[0079] The above description is only preferred embodiments of the present application, and it should be pointed out that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A method for improving the content of crude polysaccharides in Cordyceps militaris fruiting body residues, characterized in that, The method comprises the following steps: (1) cutting fresh Cordyceps militaris fruiting bodies to obtain Cordyceps militaris fruiting body segments; (2) mixing the Cordyceps militaris fruiting body segments with water, and then mixing with an extraction enzyme to perform enzyme extraction to obtain an enzyme extraction liquid; (3) adjusting the pH value of the enzyme extraction liquid to 3-5, and filtering to obtain Cordyceps militaris fruiting body residues; (4) sequentially drying and crushing the Cordyceps militaris fruiting body residues to obtain Cordyceps militaris fruiting body residue powder; (5) performing irradiation treatment on the Cordyceps militaris fruiting body residue powder; The extraction enzyme in step (2) comprises one or more of β-glucanase, xylanase and pectinase, and the amount of the extraction enzyme is 0.01%-0.5% of the dry weight of the Cordyceps militaris fruiting body segments; The temperature of the enzyme extraction in step (2) is 50-55°C, and the enzyme extraction time is 2-4h; The drying temperature in step (4) is 45-65°C, and the drying is performed until the residue moisture is 10%-15%; The irradiation treatment described in step (5) is performed using... 60 The γ-ray treatment is performed using a Co irradiation device, and the irradiation dose during the irradiation process is 4~18 kGy.

2. The method for improving the content of crude polysaccharide in Cordyceps militaris fruiting body residue according to claim 1, characterized in that, The length of the Cordyceps militaris fruiting body segments in step (1) is 1-5cm.

3. The method for increasing the content of crude polysaccharides in Cordyceps militaris fruiting body residues according to claim 1 or 2, characterized in that, The mixing mass ratio of the Cordyceps militaris fruiting body segments to water in step (2) is 1:3-5.

4. The method for improving the content of crude polysaccharide in Cordyceps militaris fruiting body residue according to claim 3, characterized in that, The component for adjusting the pH value in step (3) is an edible acid, and the edible acid comprises one or more of citric acid, malic acid, gluconic acid, lactic acid, tartaric acid, phosphoric acid, fumaric acid, adipic acid and succinic acid.

5. The method for improving the content of crude polysaccharide in Cordyceps militaris fruiting body residue according to claim 4, characterized in that, The crushing in step (4) is first coarse crushing, and then ultrafine crushing, the coarse crushing has a mesh number of 60-80 meshes, and the ultrafine crushing has a mesh number of 150-500 meshes.

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