CN102835251B - Submerged fermentation culturing method for medicinal hericium erinaceus mycelium liquid - Google Patents
Submerged fermentation culturing method for medicinal hericium erinaceus mycelium liquid Download PDFInfo
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Abstract
The invention discloses a submerged fermentation culturing method for a medicinal hericium erinaceus mycelium liquid; compared with a common edible hericium erinaceus, the obtained hericium erinaceus mycelium contains five cancer-fighting substances, seven vitamins, three active cellulases, 18 amino acids, trace elements essential to the human body and on the like except for polysaccharide and polypeptide, wherein contents of cystine, histidine and arginine with larger medicinal values are large; and the 6-7 amino acid identifying points and the protein content (16.65%) of the obtained hericium erinaceus mycelium are respectively 3 times and 1.8 times larger than those of other hericium erinaceus. Infrared spectrum also proves that the hericium erinaceus mycelium also contains a special cancer-fighting organic acid, which is not available in the common hericium erinaceus, except for a hydroxyl ammonia compound. Compared with the traditional manual solid culturing method, the submerged fermentation culturing method disclosed by the invention has the advantages as follows: the breakthrough from manual culture of the hericium erinaceus mycelium to standard and industrial production of the hericium erinaceus mycelium is achieved, the production period is reduced from the 52 days of the traditional culturing method to 14 days, energy is saved and consumption is reduced by 30%, and production cost is reduced by 70%.
Description
Technical field
The present invention relates to microorganism and cultivate technical field, the mycelial cultural method of especially medicinal Hericium erinaceus.
Background technology
Hericium erinaceus, to gain the name because its fruit body shape exactly likes the head of little monkey, it is a kind of famous and precious edible mushroom that has edible and medical value concurrently, property is put down, distinguishes the flavor of sweet, energy sharp the five internal organs, aid digestion, nourishing, anticancer, treat neurasthenia, is used for the treatment of disease and the tumour of the digestive systems such as indigestion, stomach ulcer, cancer of the esophagus, cancer of the stomach, duodenal cancer, cardiac cancer.Hericium erinaceus has been successfully applied in existing product at present, and as commercially available HOUTOU JIANWEI JIAONANG, wherein Hericium erinaceus is the Main Ingredients and Appearance of this product.
The production ways of Hericium erinaceus mainly contains three kinds at present: 1, artificial cultivation, and advantage is simple, cost is low; Shortcoming is that the cultivation cycle is long, approximately 3 months; 2, solid fermentation, cultivate the Hericium erinaceus mycelium with solid culture medium, and to obtain secondary metabolite, its advantage is: the simple and wide material sources of medium, low price; Fermentation process does not generally need strict sterile working; Incubation oxygen supply and temperature can adopt direct force air to ventilate to control; The processing of fermentation residue is simple, directly as feed or fertilizer; Shortcoming is: the medium water activity is low, the inhomogeneous and difficult stirring of medium, the growth of thalline, all inhomogeneous to the secretion of the absorption of nutriment and metabolite, make the monitoring of fermentation parameter and control difficulty, thus continued operation and automation very difficult; The temperature of the heat that microbial respiratory and metabolism produce is controlled very difficult; Adopt solid-state cultivation mostly based on empirical data and operating experience, easily pollution microbes; 3, liquid deep layer fermenting, produce a large amount of mycelium and metabolite within a short period of time to obtain optimum operation condition, advantage is: can carry out industrialization and produce continuously, cultivate raw material sources extensive, reach optimal culture condition by the controlled fermentation device, high with fugitive rate of the cycle of obtaining; Shortcoming is the cycle or longer, and mycelial output is low, is difficult to form large-scale application.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, propose a kind of medicinal Hericium erinaceus mycelium liquid submerged culturing method for making, the method can be produced the medicinal Hericium erinaceus mycelium of better quality, and method is more perfect.
In order to realize the foregoing invention purpose, the invention provides following technical scheme: the mycelial cultural method of a kind of medicinal Hericium erinaceus, comprise the following steps: the percentage of following substances is percetage by weight,
(1). the optimization screening of bacterial classification and fruit body are separated:
Strong, the fruit body that physically well develops, do not have damage by disease and insect, seed not to be scattered of growth in wild Hericium erinaceus in selecting and purchasing Zhongtiao Shan Mountain virgin forest;
At transfer room more than 100 grades in superclean bench, to in the alcohol of hericium erinaceus fruiting body immersion 75%, sterilize 1～2 minute, clean with distilled water flushing after taking-up, with antiseptic gauze, dry again, fruit body is placed on culture dish, with bell-jar or similar apparatus, cover, room temperature remains on 20 ℃, fruit body is slowly grown, after spending 2～3 days, seed sheds, fall into culture dish, then dip with tweezers the seed fallen, be inoculated in by method of scoring on the inclined-plane of test tube and cultivate, cultivating 4～5 days seeds under 22～25 ℃ can sprout, when mycelia is covered with inclined-plane substantially, the selection look white, the substrate mycelium prosperity is sturdy, the bacterium bundle is significantly excellent species, insert shaking table bottle graft kind standby,
In culture dish, medium is composed as follows:
The water that adds surplus, 6～7.5,118～121 ℃ of sterilizings of pH value 25～30 minutes;
(2). the cultivation of bacterial classification
2.1 preparation medium
The water that adds surplus, pH is 6～7.5,118～121 ℃ of sterilizings 28～32 minutes;
2.2 before inoculation, inoculation mouthful should first be sterilized by conventional method, wipe examination with the rayon balls of 75% alcohol-pickled mistake, rear unlatching ozone generator, make its air outlet all the time facing to inoculation mouthful, form a local aseptic area on every side in inoculation mouthful, then with sterile nontoxic rubber pipe box in the inoculation mouth of pipe on, to access in tank through the shaking table bottle bacterial classification of spore separation, inoculum concentration is 9%～11%, and all processes all carries out under aseptic condition, open agitator after inoculation, 100 rev/mins of mixing speeds, 25 ℃ of constant temperature, cultivate 3～4 days; The microscopy mycelia is dendroid, and without miscellaneous bacteria, outward appearance has the Archon shape;
(3). deep liquid is cultivated:
3.1 seed tank culture
Medium is following the composition:
The water that adds surplus, 118～121 ℃ of sterilizings 28～32 minutes;
The cultured bacterial classification of step (2) is seeded to seeding tank, 25～27 ℃, cultivate 3～4 days, to result be:
1.. microscopy: mycelia is dendroid, without miscellaneous bacteria;
2.. outward appearance: more white pelletoid is arranged in liquid;
3.2 the secondary tank is cultivated
Medium is following the composition:
The water that adds surplus, 118～121 ℃ of sterilizings 28～32 minutes, be seeded to the secondary tank by the cultured bacterial classification of step 3.1,25～27 ℃, cultivates 2～3 days;
3.3 three grades of tanks are cultivated
Medium is following the composition:
The water that adds surplus, 118～121 ℃ of sterilizings 28～32 minutes, be seeded to three grades of tanks by the cultured bacterial classification of step 3.2, cultivates 3～4 days for 25～27 ℃, to culture fluid, is brown, is full of mycelia, pH value 4.5 left and right, residual sugar 0.2% left and right, obtain hedgehog fungus mycelium.
Preferably, three grades of hedgehog fungus myceliums that tank obtains, with the spray drying tower of 8～10 meters, 118～121 ℃ of atomized dryings, obtain the Hericium erinaceus powder.
Compared with prior art, the present invention selects the wild Hericium erinaceus in the Zhongtiao Shan Mountain virgin forest to optimize through for many years the special medicinal white monkey bacterial classification of improveing and cultivating, and adopts liquid submerged femrentation culturing method.The present invention adopts totally-enclosed liquid deep layer fermenting technology to cultivate hericium mycelium, with the Traditional Man plate method, compare, not only realized that the production of hericium mycelium is by manual breakthrough of cultivating to standardization, suitability for industrialized production, and the production cycle shortens to 14 days in 52 days by traditional cultural method, save energy and reduce the cost 30%, production cost reduces by 70%.
The Hericium erinaceus produced with the method is except containing the polysaccharide polypeptide, still also contain five kinds different " callose ", the various saccharides combinations such as same glucose, mannitol and galactose, immunocompetence is high, certain antitumaous effect is arranged, contain VitAVitE, Cobastab
1, Cobastab
2, Cobastab
6, Cobastab
12, seven kinds of vitamins such as vitamin C, contain cellulase, hemicellulase, amylase, guaiacolase and polyphenol oxidase isoreactivity cellulase, and contain leucine, isoleucine, valine, lysine, threonine, methionine, tryptophan, phenyl alanine, histidine, arginine, alanine, asparatate, cystine, glutamic acid, proline, tyrosine, glycine, the trace element of 18 seed amino acids such as serine and needed by human etc., the larger cystine of medical value wherein, histidine, arginine content is more, amino acid is differentiated 6～7 points, protein content 16.65%, respectively 3 times and 1.8 times of other Hericium erinaceus.Also detect and confirm by infrared spectrum, except containing the hydroxyl ammoniate, also contain the special anticancer organic acid do not had in a kind of general hedgehog hydnum.
Embodiment
Below in conjunction with specific embodiment, describe the present invention, the description of this part is only exemplary and explanatory, should any restriction not arranged to protection scope of the present invention.
Embodiment 1
A kind of medicinal Hericium erinaceus mycelium liquid submerged culturing method for making, comprise the following steps: the percentage of following substances is percetage by weight,
(1). the optimization screening of bacterial classification and fruit body are separated:
Strong, the fruit body that physically well develops, do not have damage by disease and insect, seed not to be scattered of growth in wild Hericium erinaceus in selecting and purchasing Zhongtiao Shan Mountain virgin forest;
At transfer room more than 100 grades in superclean bench, by sterilization in the alcohol of hericium erinaceus fruiting body immersion 75% 2 minutes, clean with distilled water flushing after taking-up, with antiseptic gauze, dry again, fruit body is placed on culture dish, with bell-jar or similar apparatus, cover, room temperature remains on 20 ℃, fruit body is slowly grown, after spending 2 days, seed sheds, fall into culture dish, then dip with tweezers the seed fallen, be inoculated in by method of scoring on the inclined-plane of test tube and cultivate, cultivating 4 days seeds under 25 ℃ can sprout, when mycelia is covered with inclined-plane substantially, the selection look white, the substrate mycelium prosperity is sturdy, the bacterium bundle is significantly excellent species, insert shaking table bottle graft kind standby,
In culture dish, medium is composed as follows:
The water that adds surplus, pH nature, 121 ℃ of sterilizings 30 minutes;
(2). the cultivation of bacterial classification
2.1 preparation medium
The water that adds surplus, pH nature, 121 ℃ of sterilizings 30 minutes;
2.2 before inoculation, inoculation mouthful should first be sterilized by conventional method, wipe examination with the rayon balls of 75% alcohol-pickled mistake, rear unlatching ozone generator, make its air outlet all the time facing to inoculation mouthful, form a local aseptic area on every side in inoculation mouthful, then with sterile nontoxic rubber pipe box in the inoculation mouth of pipe on, to access in tank through the shaking table bottle bacterial classification of spore separation, inoculum concentration is 10%, and all processes all carries out under aseptic condition, open agitator after inoculation, 100 rev/mins of mixing speeds, 25 ℃ of constant temperature, cultivate 4 days; The microscopy mycelia is dendroid, and without miscellaneous bacteria, outward appearance has the Archon shape;
(3). deep liquid is cultivated:
3.1 seed tank culture
Medium is following the composition:
The water that adds surplus, 121 ℃ of sterilizings 30 minutes;
The cultured bacterial classification of step (2) is seeded to seeding tank, cultivates 3 days for 26 ℃, to result be:
1.. microscopy: mycelia is dendroid, without miscellaneous bacteria;
2.. outward appearance: more white pelletoid is arranged in liquid;
3.2 the secondary tank is cultivated
Medium is following the composition:
The water that adds surplus, 121 ℃ of sterilizings 30 minutes, be seeded to the secondary tank by the cultured bacterial classification of step 3.1,27 ℃, cultivates 2 days;
3.3 three grades of tanks are cultivated
Medium is following the composition:
The water that adds surplus, 121 ℃ of sterilizings 30 minutes, be seeded to three grades of tanks by the cultured bacterial classification of step 3.2, cultivates 4 days for 25 ℃, to culture fluid, is brown, is full of mycelia, pH value 4.5 left and right, residual sugar 0.2% left and right, obtain hedgehog fungus mycelium.
Three grades of hedgehog fungus myceliums that tank obtains, with the spray drying tower of 10 meters, 120 ℃ of atomized dryings, obtain the Hericium erinaceus powder.
Embodiment 2
A kind of medicinal Hericium erinaceus mycelium liquid submerged culturing method for making, comprise the following steps: the percentage of following substances is percetage by weight,
(1). the optimization screening of bacterial classification and fruit body are separated:
Strong, the fruit body that physically well develops, do not have damage by disease and insect, seed not to be scattered of growth in wild Hericium erinaceus in selecting and purchasing Zhongtiao Shan Mountain virgin forest;
At transfer room more than 100 grades in superclean bench, by sterilization in the alcohol of hericium erinaceus fruiting body immersion 75% 1 minute, clean with distilled water flushing after taking-up, with antiseptic gauze, dry again, fruit body is placed on culture dish, with bell-jar or similar apparatus, cover, room temperature remains on 20 ℃, fruit body is slowly grown, after spending 3 days, seed sheds, fall into culture dish, then dip with tweezers the seed fallen, be inoculated in by method of scoring on the inclined-plane of test tube and cultivate, cultivating 5 days seeds under 22 ℃ can sprout, when mycelia is covered with inclined-plane substantially, the selection look white, the substrate mycelium prosperity is sturdy, the bacterium bundle is significantly excellent species, insert shaking table bottle graft kind standby,
In culture dish, medium is composed as follows:
The water that adds surplus, 7.5,118 ℃ of sterilizings of pH value 30 minutes;
(2). the cultivation of bacterial classification
2.1 preparation medium
The water that adds surplus, pH is 6,121 ℃ of sterilizings 28 minutes;
2.2 before inoculation, inoculation mouthful should first be sterilized by conventional method, wipe examination with the rayon balls of 75% alcohol-pickled mistake, rear unlatching ozone generator, make its air outlet all the time facing to inoculation mouthful, form a local aseptic area on every side in inoculation mouthful, then with sterile nontoxic rubber pipe box in the inoculation mouth of pipe on, to access in tank through the shaking table bottle bacterial classification of spore separation, inoculum concentration is 11%, and all processes all carries out under aseptic condition, open agitator after inoculation, 100 rev/mins of mixing speeds, 25 ℃ of constant temperature, cultivate 3 days; The microscopy mycelia is dendroid, and without miscellaneous bacteria, outward appearance has the Archon shape;
(3). deep liquid is cultivated:
3.1 seed tank culture
Medium is following the composition:
The water that adds surplus, 118 ℃ of sterilizings 32 minutes;
The cultured bacterial classification of step (2) is seeded to seeding tank, 27 ℃, cultivate 3 days, to result be:
1.. microscopy: mycelia is dendroid, without miscellaneous bacteria;
2.. outward appearance: more white pelletoid is arranged in liquid;
3.2 the secondary tank is cultivated
Medium is following the composition:
The water that adds surplus, 121 ℃ of sterilizings 28 minutes, be seeded to the secondary tank by the cultured bacterial classification of step 3.1,25 ℃, cultivates 3 days;
3.3 three grades of tanks are cultivated
Medium is following the composition:
The water that adds surplus, 118 ℃ of sterilizings 32 minutes, be seeded to three grades of tanks by the cultured bacterial classification of step 3.2, cultivates 3 days for 26 ℃, to culture fluid, is brown, is full of mycelia, pH value 4.5 left and right, residual sugar 0.2% left and right, obtain hedgehog fungus mycelium.
Embodiment 3
A kind of medicinal Hericium erinaceus mycelium liquid submerged culturing method for making, comprise the following steps: the percentage of following substances is percetage by weight,
(1). the optimization screening of bacterial classification and fruit body are separated:
Strong, the fruit body that physically well develops, do not have damage by disease and insect, seed not to be scattered of growth in wild Hericium erinaceus in selecting and purchasing Zhongtiao Shan Mountain virgin forest;
At transfer room more than 100 grades in superclean bench, by sterilization in the alcohol of hericium erinaceus fruiting body immersion 75% 2 minutes, clean with distilled water flushing after taking-up, with antiseptic gauze, dry again, fruit body is placed on culture dish, with bell-jar or similar apparatus, cover, room temperature remains on 20 ℃, fruit body is slowly grown, after spending 2 days, seed sheds, fall into culture dish, then dip with tweezers the seed fallen, be inoculated in by method of scoring on the inclined-plane of test tube and cultivate, cultivating 4 days seeds under 25 ℃ can sprout, when mycelia is covered with inclined-plane substantially, the selection look white, the substrate mycelium prosperity is sturdy, the bacterium bundle is significantly excellent species, insert shaking table bottle graft kind standby,
In culture dish, medium is composed as follows:
The water that adds surplus, 6,121 ℃ of sterilizings of pH value 25 minutes;
(2). the cultivation of bacterial classification
2.1 preparation medium
The water that adds surplus, pH is 7.5,121 ℃ of sterilizings 28 minutes;
2.2 before inoculation, inoculation mouthful should first be sterilized by conventional method, wipe examination with the rayon balls of 75% alcohol-pickled mistake, rear unlatching ozone generator, make its air outlet all the time facing to inoculation mouthful, form a local aseptic area on every side in inoculation mouthful, then with sterile nontoxic rubber pipe box in the inoculation mouth of pipe on, to access in tank through the shaking table bottle bacterial classification of spore separation, inoculum concentration is 9%, and all processes all carries out under aseptic condition, open agitator after inoculation, 100 rev/mins of mixing speeds, 25 ℃ of constant temperature, cultivate 4 days; The microscopy mycelia is dendroid, and without miscellaneous bacteria, outward appearance has the Archon shape;
(3). deep liquid is cultivated:
3.1 seed tank culture
Medium is following the composition:
The water that adds surplus, 121 ℃ of sterilizings 28 minutes;
The cultured bacterial classification of step (2) is seeded to seeding tank, 25 ℃, cultivate 4 days, to result be:
1.. microscopy: mycelia is dendroid, without miscellaneous bacteria;
2.. outward appearance: more white pelletoid is arranged in liquid;
3.2 the secondary tank is cultivated
Medium is following the composition:
The water that adds surplus, 118 ℃ of sterilizings 32 minutes, be seeded to the secondary tank by the cultured bacterial classification of step 3.1,27 ℃, cultivates 2 days;
3.3 three grades of tanks are cultivated
Medium is following the composition:
The water that adds surplus, 121 ℃ of sterilizings 28 minutes, be seeded to three grades of tanks by the cultured bacterial classification of step 3.2, cultivates 4 days for 25 ℃, to culture fluid, is brown, is full of mycelia, pH value 4.5 left and right, residual sugar 0.2% left and right, obtain hedgehog fungus mycelium.
Above-described embodiment 2 and the prepared hedgehog fungus mycelium of embodiment 3, can adopt drying mode as described in Example 1 to obtain the Hericium erinaceus powder equally.
Detect the hedgehog fungus mycelium that above-described embodiment 1 to embodiment 3 obtains, result is compared with general edible Hericium erinaceus, except containing the polysaccharide polypeptide, still also contain five kinds different " callose ", the various saccharides combinations such as same glucose, mannitol and galactose, immunocompetence is high, and certain antitumaous effect is arranged, and contains VitAVitE, Cobastab
1, Cobastab
2, Cobastab
6, Cobastab
12, seven kinds of vitamins such as vitamin C, contain cellulase, hemicellulase, amylase, guaiacolase and polyphenol oxidase isoreactivity cellulase, and contain leucine, isoleucine, valine, lysine, threonine, methionine, tryptophan, phenyl alanine, histidine, arginine, alanine, asparatate, cystine, glutamic acid, proline, tyrosine, glycine, the trace element of 18 seed amino acids such as serine and needed by human etc., the larger cystine of medical value wherein, histidine, arginine content is more, amino acid is differentiated 6～7 points, protein content 16.65%, respectively 3 times and 1.8 times of other Hericium erinaceus.Also detect and confirm by infrared spectrum, except containing the hydroxyl ammoniate, also contain the special anticancer organic acid do not had in a kind of general hedgehog hydnum.
Simultaneously the production cycle of the various embodiments described above is 14 ± 2 days, more than the cycle of 52 days of traditional cultural method short, energy-saving and cost-reducing 30%, production cost reduces by 70%.
It is only below the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (2)
1. a medicinal Hericium erinaceus mycelium liquid submerged culturing method for making, comprise the following steps: the percentage of following substances is percetage by weight,
(1). the optimization screening of bacterial classification and fruit body are separated:
Strong, the fruit body that physically well develops, do not have damage by disease and insect, seed not to be scattered of growth in wild Hericium erinaceus in selecting and purchasing Zhongtiao Shan Mountain virgin forest;
At transfer room more than 100 grades in superclean bench, to in the alcohol of hericium erinaceus fruiting body immersion 75%, sterilize 1～2 minute, clean with distilled water flushing after taking-up, with antiseptic gauze, dry again, fruit body is placed on culture dish, with the bell-jar apparatus, cover, room temperature remains on 20 ℃, fruit body is slowly grown, after spending 2～3 days, seed sheds, fall into culture dish, then dip with tweezers the seed fallen, be inoculated in by method of scoring on the inclined-plane of test tube and cultivate, cultivating 4～5 days seeds under 22～25 ℃ can sprout, when mycelia is covered with inclined-plane substantially, the selection look white, the substrate mycelium prosperity is sturdy, the bacterium bundle is significantly excellent species, insert shaking table bottle graft kind standby,
In culture dish, medium is composed as follows:
The water that adds surplus, 6～7.5,118～121 ℃ of sterilizings of pH value 25～30 minutes;
(2). the cultivation of bacterial classification
2.1 preparation medium
The water that adds surplus, pH is 6～7.5,118～121 ℃ of sterilizings 28～32 minutes;
2.2 before inoculation, inoculation mouthful should first be sterilized by conventional method, wipe examination with the rayon balls of 75% alcohol-pickled mistake, rear unlatching ozone generator, make its air outlet all the time facing to inoculation mouthful, form a local aseptic area on every side in inoculation mouthful, then with sterile nontoxic rubber pipe box in the inoculation mouth of pipe on, to access in tank through the shaking table bottle bacterial classification of spore separation, inoculum concentration is 9%～11%, and all processes all carries out under aseptic condition, open agitator after inoculation, 100 rev/mins of mixing speeds, 25 ℃ of constant temperature, cultivate 3～4 days; The microscopy mycelia is dendroid, and without miscellaneous bacteria, outward appearance has the Archon shape;
(3). deep liquid is cultivated:
3.1 seed tank culture
Medium is following the composition:
The water that adds surplus, 118～121 ℃ of sterilizings 28～32 minutes;
The cultured bacterial classification of step (2) is seeded to seeding tank, 25～27 ℃, cultivate 3～4 days, to result be:
1.. microscopy: mycelia is dendroid, without miscellaneous bacteria;
2.. outward appearance: more white pelletoid is arranged in liquid;
3.2 the secondary tank is cultivated
Medium is following the composition:
The water that adds surplus, 118～121 ℃ of sterilizings 28～32 minutes, be seeded to the secondary tank by the cultured bacterial classification of step 3.1,25～27 ℃, cultivates 2～3 days;
3.3 three grades of tanks are cultivated
Medium is following the composition:
The water that adds surplus, 118～121 ℃ of sterilizings 28～32 minutes, be seeded to three grades of tanks by the cultured bacterial classification of step 3.2, cultivates 3～4 days for 25～27 ℃, to culture fluid, is brown, is full of mycelia, pH value 4.5 left and right, residual sugar 0.2% left and right, obtain hedgehog fungus mycelium.
2. cultural method as claimed in claim 1, it is characterized in that: three grades of hedgehog fungus myceliums that tank obtains, with the spray drying tower of 8～10 meters, 118～121 ℃ of atomized dryings, obtain the Hericium erinaceus powder.
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| CN106376354A (en) * | 2016-08-29 | 2017-02-08 | 浙江省医学科学院 | Hericium erinaceus polysaccharide extraction raw material preparation method |
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| CN109287381A (en) * | 2018-11-30 | 2019-02-01 | 成都市农林科学院 | A kind of Hericium erinaceus culture substrate and the method using the substrate fermentation culture Hericium erinaceus mycoplasma |
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