CN108753625B - Polysaccharide-producing space Hericium coralloides ST21-3 and application thereof in improving biological immune activity - Google Patents
Polysaccharide-producing space Hericium coralloides ST21-3 and application thereof in improving biological immune activity Download PDFInfo
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- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
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- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
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Abstract
The invention discloses space coralloid hericium erinaceus ST21-3 for producing polysaccharide and application thereof in improving biological immunocompetence. The preservation number of the coralloid Hericium erinaceus (Hericium coralloides) Fullarton-H-ST21-3 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 15379. The invention adopts space spacecraft of Tiangong No. 2 and Shenzhou No. 11 to carry hericium erinaceus which are returned to the ground and subjected to space mutagenesis, takes the original hericium erinaceus on the ground as a contrast, and breeds a strain with higher immunocompetence, thereby providing practical basis for the application of space hericium erinaceus mycelium polysaccharide in improving biological immunocompetence. Meanwhile, the invention widens the application range of space microorganisms to a certain extent and fills the gap of the research and development of the microbial functions of food space fungi.
Description
Technical Field
The invention relates to space coralloid hericium erinaceus ST21-3 for producing polysaccharide and application thereof in improving biological immunocompetence.
Background
Hericium Erinaceus Polysaccharides (HEP) are composed of glucose, galactose and mannose, are glucans formed by main chains connected through beta- (1, 3) glycosidic bonds and branched chains connected through beta- (1, 6) glycosidic bonds, and are important functional active substances produced by fermentation of Hericium erinaceus. The Hericium erinaceus polysaccharide comprises polysaccharide in mycelium cells and polysaccharide in fermentation liquor. The Hericium erinaceus mycelium polysaccharide has the biological activities of reducing blood sugar, reducing blood lipid, resisting tumor, resisting oxidation, etc., in addition to the immunoregulation activity. The Hericium erinaceus polysaccharide immunoregulation activity is shown in the following steps: can improve the activity of natural killer cells, promote the generation of delayed hypersensitivity (anti-allergic reaction), enhance the phagocytic function of macrophages, and has no obvious effect on humoral immunity and proliferation of immune organs. This indicates that the hericium erinaceus mycelium polysaccharide mainly regulates the immune function of the organism by enhancing the activity of immune cells. In addition, many researches prove that the hericium erinaceus polysaccharide has important functions in the aspects of treating gastritis and peptic ulcer, improving the hypoxia tolerance of organisms, increasing the cardiac blood output, accelerating the blood circulation of the organisms and the like, and is a functional bacterium for both food and medicine.
Currently, domestic and foreign scholars mainly focus on optimization of polysaccharide substance extraction conditions and analysis of contained substance structures, research on immunomodulatory activity of polysaccharide substances is still in the beginning stage, research on space microorganisms mostly stays in application to space pathogenic bacteria, saprophytic bacteria and pharmacy, and development of food space microorganisms is still in the sprouting stage.
Disclosure of Invention
The invention aims to provide space coralloid hericium erinaceus ST21-3 capable of producing polysaccharide and application thereof in improving biological immune activity.
The coralloid Hericium erinaceus (Hericium coralloides) Fularton-H-ST 21-3 is preserved in China general microbiological culture Collection center (CGMCC for short; address: No. 3 Hospital No.1 of North West Lu of the Chaoyang district, China academy of sciences, postal code: 100101) in 24 months and 4 in 2018, and the preservation number is CGMCC NO. 15379.
The invention also protects the application of the coralloid Hericium (Hericium coralloides) Fullarton-H-ST21-3 or the mycelium thereof in preparing products; the application of the product is (1) and/or (2) as follows:
(1) increasing the activity of immune cells;
(2) improving immunity of organism.
The invention also provides a preparation method of the hericium erinaceus mycelium polysaccharide, which comprises the following steps: culturing Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 to obtain mycelium; extracting polysaccharide from mycelium to obtain the coralloid hericium erinaceus mycelium polysaccharide.
In the method, the Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 is cultured by using a seed fermentation medium.
In the method, the culture condition is shaking culture at 28 ℃ and 160 r/min.
The method specifically comprises the following steps:
(a) inoculating Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 into seed fermentation medium for culturing;
(b) after the step (a) is finished, transferring the culture into a seed fermentation culture medium according to the inoculation amount of 10% (volume percentage) for culture, and collecting mycelia; extracting polysaccharide from mycelium.
In the step (a), the culture condition is shaking culture at 28 ℃ and 160r/min, and the culture time is 12 days.
In the step (b), the culture condition is shaking culture at 28 ℃ and 160r/min, and the culture time is 5 days.
In the method, the method for collecting the mycelium specifically comprises the following steps: filtering the culture system with three layers of sterile gauze, and collecting mycelium (washing with sterile distilled water for 3-4 times, and vacuum freeze drying).
In the method, the method for extracting the polysaccharide from the mycelium specifically comprises the following steps: freeze-drying mycelium, crushing the mycelium into powder, mixing the powder with distilled water, extracting the mixture for 2 hours in a boiling water bath, filtering the mixture by three layers of gauze, collecting an extracting solution (the extracting solution can be combined after residues are repeatedly extracted), carrying out alcohol precipitation on the extracting solution (the condition can be that the extracting solution is placed at 4 ℃ for 12 hours), collecting precipitate, and drying the precipitate (constant temperature drying at 60 ℃ or vacuum freeze drying can be adopted) to obtain the polysaccharide.
The invention also protects the hericium erinaceus mycelium polysaccharide obtained by any one of the methods.
The invention also protects the application of the hericium erinaceus mycelium polysaccharide in preparing products; the application of the product is (1) and/or (2) as follows:
(1) increasing the activity of immune cells;
(2) improving immunity of organism.
The invention also provides a product, the active component of which is Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 or mycelium thereof; the application of the product is (1) and/or (2) as follows:
(1) increasing the activity of immune cells;
(2) improving immunity of organism.
The invention also protects a product, the active component of which is the hericium erinaceus mycelium polysaccharide; the application of the product is (1) and/or (2) as follows:
(1) increasing the activity of immune cells;
(2) improving immunity of organism.
The invention also provides a kit for preparing the Hericium erinaceus mycelium polysaccharide, which comprises Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 and a seed fermentation culture medium.
Any of the above products can be specifically food, medicine or health care product and the like.
Any one of the seed fermentation culture media consists of a solute and a solvent; the solutes and their concentrations in the seed fermentation medium were as follows: 15-25g/L of soluble starch, 25-35g/L of glucose, 5-15g/L of yeast extract powder, 0.5-0.7g/L of magnesium sulfate heptahydrate and 2.5-3.5g/L of potassium dihydrogen phosphate; the solvent is water.
The concentrations in the seed fermentation medium and in the seed fermentation medium are as follows: 20g/L of soluble starch, 30g/L of glucose, 10g/L of yeast extract powder, 0.6g/L of magnesium sulfate heptahydrate and 3.0g/L of potassium dihydrogen phosphate.
The solvent in any one of the above seed fermentation media may be distilled water.
Any of the above-described methods for increasing the activity of an immune cell may be specifically characterized by increasing the amount of NO released from macrophages.
The macrophage may specifically be RAW264.7 cells.
The invention adopts space craft No. 2 and 11, which are carried by spacecraft of Tiangong and 2 and returned to ground and mutagenized by space, and the Hericium erinaceus has certain biological properties (such as individual shape, colony characteristics, physiological and biochemical characteristics, immunocompetence and the like) and fermentation production performance (such as biomass, product quantity, enzyme activity, titer, fermentation speed and the like) and changes the gene mutation frequency to a certain extent so as to change the functional activity due to the mutagenic actions of space microgravity effect, high vacuum, extreme temperature difference, weak magnetic field, high-energy particle (electron, proton and heavy ion) radiation and the like.
According to the invention, the original hericium erinaceus on the ground is used as a reference, and the strain with higher immunocompetence is bred, so that a practical basis is provided for the application of the spatial hericium erinaceus mycelium polysaccharide in the improvement of the biological immunocompetence. Meanwhile, the invention widens the application range of space microorganisms to a certain extent and fills the gap of the research and development of the microbial functions of food space fungi.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
Ground Hericium erinaceus (Hericium erinaceus) GT 21: china center for industrial microorganism preservation, number 14026, abbreviated as ground Hericium Erinaceus GT 21.
YEPD medium: 10g of yeast extract powder, 20g of peptone, 20g of glucose, 17g of agar and distilled water to a constant volume of 1000mL, and the pH value is 6.0.
Seed fermentation culture medium: 20g of soluble starch, 30g of glucose, 10g of yeast extract powder, 0.6g of magnesium sulfate heptahydrate, 3g of monopotassium phosphate and distilled water to a constant volume of 1000mL, and the pH value is natural.
The macrophages used in the examples were RAW264.7 cells, purchased from ATCC, usa.
Example 1 screening of spatial Hericium Erinaceus
Space mutagenesis of bacterial strains
The ground hericium erinaceus GT21 is carried back by spacecrafts of Tiangong No. 2 and Shenzhou No. 11 to obtain a plurality of space mutation strains.
Secondly, activation of the strains
Taking a space mutation strain frozen in a-80 ℃ ultra-low temperature refrigerator and a ground hericium erinaceus GT21 glycerol tube storage strain, dipping a small amount of bacterial liquid by an inoculating ring, streaking, inoculating on a YEPD culture medium test tube inclined plane, culturing for 6 days at 28 ℃, continuously activating for 3 generations, and then using for subsequent tests.
Third, the enlargement culture of the bacterial strains
Inoculating the space mutation strain activated in the second step for 3 generations and ground hericium GT21 into a YEPD culture medium triangular flask inclined plane, and culturing at 28 ℃ for 6 days.
Fourthly, separation and purification of bacterial strains
And after the third step is finished, taking the space mutation strain and the ground hericium erinaceus GT21 which grow well on the inclined plane of the YEPD culture medium triangular flask, washing surface spores with 100mL of sterile normal saline, beating for 1min by using a sterile homogenizer according to 8 times/sec to fully separate the spores from the mycelia, and filtering by using three layers of sterile gauze to obtain a pure spore suspension. Taking 1mL of spore suspension, diluting with 9mL of sterile physiological saline by 10-fold gradient, and taking 10-2The spores were diluted and plated on YEPD medium plates and incubated at 28 ℃ for 6 days. Observing the colonial morphology of space mutation strain and ground Hericium Erinaceus GT21, and selecting bacteriaThe space mutation strain single colony with the colony characteristic greatly different from the ground strain is purified and cultured on a YEPD culture medium test tube inclined plane and cultured for 4 days at 28 ℃ to obtain the strains with the numbers of ST21-1 to ST 21-10.
Fifthly, re-screening of strains
1. Preparation of mycelia
Inoculating a strain to be tested to a seed fermentation culture medium, carrying out shaking culture at 28 ℃ and 160r/min for 12 days, beating a sterile homogenizer for 8 times/sec for 1min, beating mycelium pellets, transferring the mycelium pellets into the seed fermentation culture medium according to the inoculation amount of 10% (volume percentage), carrying out shaking culture at 28 ℃ and 160r/min for 5 days, taking a fermentation system, filtering by using three layers of sterile gauze, collecting mycelia, washing the obtained mycelia with sterile distilled water for 3-4 times, and carrying out vacuum freeze drying.
2. Preparation of mycelium crude polysaccharide
Grinding the freeze-dried mycelia in the step 1 into powder, weighing 1g of a sample, adding 18mL of distilled water, extracting for 2h in a boiling water bath, filtering with three layers of gauze, repeatedly extracting filter residues for 1 time, combining extracting solutions, fixing the volume of the extracting solution in 20mL of distilled water, adding 300mL of absolute ethyl alcohol, shaking up, standing at 4 ℃ for 12h, centrifuging at 10000r/min for 20min, collecting precipitates, centrifuging and washing with 95% of ethyl alcohol for 2 times (the centrifugation conditions are the same as above), and drying the precipitates at the constant temperature of 60 ℃ until the mass is constant, thereby obtaining the crude mycelia polysaccharide.
3. Reagent preparation
(1) Preparing a double-antibody solution: 3mg of penicillin solid powder was dissolved in 100mL of distilled water to prepare a 30. mu.g/mL penicillin solution. 10mg of streptomycin solid powder was dissolved in 100mL of distilled water to prepare a 100. mu.g/mL streptomycin solution. Mixing the penicillin solution and the streptomycin solution in equal volume to obtain double-antibody solution, and storing at-20 deg.C.
(2) Preparation of PBS (phosphate buffer solution): taking 0.14mol/L NaCl, 0.0027mol/L KCl and 0.004 mol/L Na2HPO4And 0.002mol/L KH2PO4The mixture was mixed in equal volumes, adjusted to pH 7.40 with 1mol/L NaOH, sterile filtered through a 0.22 μm filter and diluted with distilled water to a PBS concentration of 10 μ g/mL for use.
(3) The kit for detecting nitric oxide Griess comprises: biyuntian corporation, stored at 4 ℃.
4. Preparation of culture medium
DMEM complete medium (volume percent): and (3) uniformly mixing 89% (volume percentage content) of DMEM high-sugar medium, 10% (volume percentage content) of FBS (fetal bovine serum) and 1% (volume percentage content) of double-antibody solution to obtain the DMEM complete medium.
DMEM high-glucose medium and FBS (fetal bovine serum) were purchased from Shanghai Biotech engineering, Inc.
5. Preparation of samples
Accurately weighing 5mg of the mycelium crude polysaccharide prepared in the step 2 in a sterile centrifuge tube, preparing a 5mg/mL polysaccharide solution by using PBS (phosphate buffer solution) with pH7.4, centrifuging for 30min at 12000r/min, collecting supernatant, transferring the supernatant into a 1.5mL sterile centrifuge tube in a sterile operation, and diluting a polysaccharide sample to 1mg/mL, 500 mu g/mL and 250 mu g/mL by using PBS with pH 7.4.
6. Preparation of macrophage suspension
Selecting macrophage in logarithmic phase, sucking out culture solution from cell culture flask, adding 1ml PBS (pH7.4) for cleaning, sucking out supernatant, adding 0.25% trypsin, placing the flask in 5% CO2The cells were suspended in an incubator at 37 ℃ for 6min, 1mL of DMEM complete medium was added to terminate the digestion reaction, and the cells were aspirated with a pipette. Transferring the cell suspension into 15mL sterile centrifuge tube, centrifuging at 1000r/min for 5min, removing supernatant, adding 1mL PBS (pH7.4) to wash the precipitate, centrifuging at 1000r/min for 5min, removing supernatant, adding DMEM complete medium to suspend the cells, counting with a blood counting cell plate, diluting the cells to 5 × 10 with DMEM complete medium5cell/mL, namely macrophage suspension.
7. Nitric Oxide (NO) concentration determination
The macrophage suspension prepared in step 6 was added to 48-well plates (purchased from Coster Co., Ltd.) in an amount of 180. mu.L, 20. mu.L of the polysaccharide samples obtained in step 5 at different concentrations were added to each well, PBS (10. mu.g/mL) at pH7.4 was used as a negative control, LPS (10. mu.g/mL bacterial lipopolysaccharide, Wawter) was used as a positive control, and the mixture was washed with water and washed with water at 37 ℃ to obtain a suspension℃、 5％CO2Culturing in an incubator for 48h to obtain macrophage treating liquid. Diluting the sodium nitrite standard substance in the Griess kit into different gradients by using a DMEM complete culture medium, respectively adding the sodium nitrite standard substance and macrophage HEP treatment solution into another 48-hole plate according to the addition amount of 200 mu L/hole, then respectively adding the Griess Reagent I and Griess Reagent II into the kit according to the addition amount of 200 mu L/hole at room temperature, and measuring the absorbance value (A value) at 540 nm. The results are shown in Table 1.
TABLE 1 Effect of mycelium polysaccharide solutions of different concentrations on NO release (μmol/L) from RAW264.7 cells
As can be seen from Table 1, the amounts of NO released from RAW264.7 macrophages stimulated by the crude hyphal polysaccharides of the ST21-1 to ST21-10 strains were significantly different and increased with the increase in the concentration of the hyphal polysaccharide solution, wherein the NO released from RAW264.7 macrophages stimulated by the crude hyphal polysaccharides of the ST21-3 strain was higher than those of the other strains at each concentration. Because the NO release amount of macrophages is in positive correlation with in vitro immune activity, hypha polysaccharide produced by the ST21-3 strain has the strongest stimulation effect on RAW264.7 macrophages, the NO release amount is 47.21 mu mol/L, which is 3.91 times of NO release amount of ground strain GT21 mycelium polysaccharide acting on RAW264.7 cells, and the ST21-3 strain has higher in vitro immune activity and can become the most valuable strain for improving biological immune activity.
Sixthly, morphological identification and molecular biological identification of ST21-3 strain
1. Colonial characteristics of the ST21-3 strain: the colony is white, slightly yellowish, felt-like, and has many irregular spherical bulges on the surface, and the aerial mycelium is wool-like. The back gradually changes from white to yellow and yellow brown from the center of the colony, and the color change gradually enlarges and deepens.
2. The 18S rDNA of the ST21-3 strain is detected, and the sequencing result is shown as the sequence in the sequence table 1. The 18s rDNA identification showed that the ST21-3 strain was more than 99% similar to Hericium coralloides. Through morphology and 18S rDNA identification, the ST21-3 strain can be determined to belong to the coralloid hericium erinaceus.
Collection of seven, ST21-3 strains
The strain ST21-3 is named as Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3, and is preserved in China general microbiological culture Collection center (CGMCC for short; address: No. 3 of West Lu No.1 of Beijing university Hokko at sunward, China academy of sciences Microbiol research, postal code: 100101) 24 days in 2018, wherein the preservation number is CGMCC NO. 15379. Hericium coralloides (Hericium coralloides)
Fullarton-H-ST21-3 is abbreviated as Hericium coralloides SST 21-3.
Eighth, genetic stability
Subculturing the Hericium coralloides SST21-3 strain, performing a genetic stability test, and determining the influence of mycelium polysaccharide solutions with different concentrations in each generation on NO release amount of macrophage RAW264.7 cells to judge the genetic stability, wherein the results are shown in Table 2.
TABLE 2 influence of the 10 th generation different concentrations of mycelium polysaccharide solutions of ST21-3 on NO release from RAW264.7 cells (. mu. mol/L)
As can be seen from Table 2, after the passage of ST21-3 to 10, the NO release amount is still at a high level, which indicates that the immunocompetence of polysaccharide solutions with different concentrations on RAW264.7 cells is at a high level, and the genetic stability of Hericium coralloides SST21-3 in the aspect of improving the immunobiological activity can be basically judged.
Example 2 measurement of polysaccharide from Hericium erinaceum mycelium ST21-3
1. Preparation of mycelia
Inoculating Hericium coralloides SST21-3 to a seed fermentation culture medium, performing shaking culture at 28 deg.C and 160r/min for 12 days, beating with an aseptic homogenizer for 8 times/sec for 1min to disperse mycelium pellets, transferring into the seed fermentation culture medium according to the inoculum size of 10% (volume percentage), performing shaking culture at 28 deg.C and 160r/min for 5 days, collecting the fermentation system, filtering with three layers of aseptic gauze, collecting mycelia, washing the obtained mycelia with aseptic distilled water for 3-4 times, and vacuum freeze-drying.
2. Preparation of mycelium crude polysaccharide
Grinding the freeze-dried mycelia in the step 1 into powder, weighing 1g of a sample, adding 18mL of distilled water, extracting for 2h in a boiling water bath, filtering with three layers of gauze, repeatedly extracting filter residues for 1 time, combining extracting solutions, fixing the volume of the extracting solution in 20mL of distilled water, adding 300mL of absolute ethyl alcohol, shaking up, standing at 4 ℃ for 12h, centrifuging at 10000r/min for 20min, collecting precipitates, and carrying out vacuum freeze drying on the precipitates to obtain crude mycelia polysaccharide.
3. Drawing of standard curve
And (3) drying standard glucose at 105 ℃ to constant weight, preparing a 10mg/mL glucose standard product mother solution, and diluting 200 times to 50 mug/mL with constant volume by using a volumetric flask, thus obtaining the glucose standard solution. Respectively sucking 0, 0.2, 0.4, 0.6, 0.8 and 1.0mL of standard glucose solution, placing the standard glucose solution in a test tube, supplementing the standard glucose solution to 1mL by distilled water, then adding 0.5mL of 5% (mass percent) phenol aqueous solution, quickly adding 2.5mL of concentrated sulfuric acid (mass fraction of 98%) after uniformly mixing, fully mixing the reaction liquid by using a vortex oscillator, then placing the test tube in a boiling water bath for reacting for 15min, cooling the water bath for 10min to room temperature, measuring the absorbance value (A value) of the test tube at 490nm by using an ultraviolet spectrophotometer, and drawing a standard curve by using the glucose mass concentration as an abscissa and the absorbance value as an ordinate. The regression equation for the standard curve is y-0.0152 x +0.0208 (R)2＝0.9995)。
4. Determination of Total sugar content
The total sugar content measured by the phenol-sulfuric acid method is the polysaccharide content. And (3) weighing 10mg of the crude polysaccharide prepared in the step (2) by using a ten-thousandth electronic balance, dissolving the crude polysaccharide in 50mL of distilled water, and then diluting the crude polysaccharide with the distilled water until the polysaccharide concentration is within the range of a standard curve to obtain a sample solution to be detected. Taking 1mL of diluted sample solution to be detected into a test tube, adding 0.5mL of 5% (mass percent) phenol solution, quickly adding 2.5mL of concentrated sulfuric acid (mass fraction of 98%) after uniformly mixing, fully mixing the reaction solution by using a vortex oscillator, then placing the test tube into a boiling water bath for reacting for 15min, cooling the test tube in the water bath for 10min to room temperature, and measuring the absorbance value (A value) of the test tube at 490nm by using an ultraviolet spectrophotometer. And substituting the measured absorbance y value into a glucose standard curve regression equation to obtain the polysaccharide content x value (mg/mL). The results of polysaccharide from Hericium coralloides ST21-3 mycelium are shown in Table 3.
Because the small molecules of the crude polysaccharide are removed by alcohol precipitation during preparation, the content of reducing sugar in the crude polysaccharide is extremely low and can be ignored.
The total content of hypha polysaccharide is equal to fermentation hypha biomass multiplied by hypha polysaccharide content/100.
TABLE 3 Hericium coralloides ST21-3 mycelium polysaccharide results
As can be seen from Table 3, the biomass of mycelium of Hericium coralloides SST21-3 was 3.52g/100mL, the polysaccharide content was 1.08mg/100mL, and the total amount of polysaccharide was 38.17mg/100 mL.
Sequence listing
<110> Fuleton biotechnology and technology (Beijing) Ltd
<120> space coralloid hericium coralloid ST21-3 for producing polysaccharide and application thereof in improving biological immune activity
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 648
<212> DNA
<213> Hericium coralloides (Hericium coralloides)
<400> 1
ccttccgtaa gggggcctgc ggaaggatca ttaatgaatt tgaaaggagt tttgttgctg 60
gcttgtcaac ccaggcatgt gcacactccg atctcatcca tcttacacct gtgcaccctt 120
gcgtgggtct gtcggctttg cggtvgttcg ggcttgcgtt ttttctataa acttttatgt 180
agtaacagaa tgtcttaaat gctataaacg catcttatac aactttcaac aacggatctc 240
ttggctctcg catcgatgaa gaacgcagcg aaatgcgata agtaatgtga attgcagaat 300
tcagtgaatc atcgaatctt tgaacgcacc ttgcgcccct tggtattccg agggcacgcc 360
tgttcgagtg tcgtgaaatt ctcaactcaa tcctcttgtt atgagagggc tgggcttgga 420
cttggaggtc ttgccggtgg ttccttcggg accgtcggct cctcttgaat gcatgagtgg 480
atcccttttt gtagggtttg cccttggtgt gataatatct acgccgcggg tagccttgcg 540
cgctggtctg cttctaaccg tccttcggga catgtttttc atctcaactt gacctcgaat 600
caggctgctg ccgctgattt gcccaagcat atcaatacgc gaaggaaa 648
Claims (7)
1. Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3, wherein the preservation number of the Hericium coralloides (Hericium coralloides) in the China general microbiological culture Collection center is CGMCC No. 15379.
2. Use of the Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 or its mycelium for the preparation of a product; the application of the product is (1) and/or (2) as follows:
(1) increasing the activity of immune cells;
(2) improving immunity of organism.
3. A preparation method of hericium erinaceus mycelium polysaccharide comprises the following steps: culturing Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 to obtain mycelium, and extracting polysaccharide from the mycelium to obtain the mycelium polysaccharide of Hericium coralloides.
4. A method as claimed in claim 3, comprising the steps of: in the method, the Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 is cultured in a seed fermentation medium;
the seed fermentation culture medium consists of solute and solvent; the solutes and their concentrations in the seed fermentation medium were as follows: 15-25g/L of soluble starch, 25-35g/L of glucose, 5-15g/L of yeast extract powder, 0.5-0.7g/L of magnesium sulfate heptahydrate and 2.5-3.5g/L of potassium dihydrogen phosphate; the solvent is water.
5. The method of claim 3 or 4, wherein: in the method, the culture condition is shaking culture at 28 ℃ and 160 r/min.
6. A product comprising as an active ingredient the Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 or a mycelium thereof according to claim 1; the application of the product is (1) and/or (2) as follows:
(1) increasing the activity of immune cells;
(2) improving immunity of organism.
7. A kit for preparing a mycelia polysaccharide of Hericium erinaceus comprising Hericium coralloides (Hericium coralloides) Fullarton-H-ST21-3 of claim 1 and the seed fermentation medium of claim 4.
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