CN112826082A - Preparation method of mushroom enzyme - Google Patents
Preparation method of mushroom enzyme Download PDFInfo
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- CN112826082A CN112826082A CN202110074079.2A CN202110074079A CN112826082A CN 112826082 A CN112826082 A CN 112826082A CN 202110074079 A CN202110074079 A CN 202110074079A CN 112826082 A CN112826082 A CN 112826082A
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- mushroom
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- 235000001674 Agaricus brunnescens Nutrition 0.000 title claims abstract description 149
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 63
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 63
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 239000001963 growth medium Substances 0.000 claims abstract description 60
- 238000000855 fermentation Methods 0.000 claims abstract description 44
- 230000004151 fermentation Effects 0.000 claims abstract description 43
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 38
- 241000894006 Bacteria Species 0.000 claims abstract description 19
- 239000004310 lactic acid Substances 0.000 claims abstract description 19
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 19
- 229910017053 inorganic salt Inorganic materials 0.000 claims abstract description 6
- 239000000843 powder Substances 0.000 claims description 79
- 239000007788 liquid Substances 0.000 claims description 64
- 239000003651 drinking water Substances 0.000 claims description 62
- 235000020188 drinking water Nutrition 0.000 claims description 62
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 59
- 238000002791 soaking Methods 0.000 claims description 51
- 240000000599 Lentinula edodes Species 0.000 claims description 42
- 235000001715 Lentinula edodes Nutrition 0.000 claims description 42
- 238000012258 culturing Methods 0.000 claims description 36
- 241000186660 Lactobacillus Species 0.000 claims description 35
- 229940039696 lactobacillus Drugs 0.000 claims description 35
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 230000001954 sterilising effect Effects 0.000 claims description 21
- 239000007787 solid Substances 0.000 claims description 19
- 238000004659 sterilization and disinfection Methods 0.000 claims description 19
- 239000011550 stock solution Substances 0.000 claims description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 15
- 239000001888 Peptone Substances 0.000 claims description 15
- 108010080698 Peptones Proteins 0.000 claims description 15
- 229940041514 candida albicans extract Drugs 0.000 claims description 15
- 239000008103 glucose Substances 0.000 claims description 15
- 235000019319 peptone Nutrition 0.000 claims description 15
- 239000012138 yeast extract Substances 0.000 claims description 15
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 13
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 13
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 12
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 238000004537 pulping Methods 0.000 claims description 12
- 238000004140 cleaning Methods 0.000 claims description 11
- 238000000227 grinding Methods 0.000 claims description 11
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 11
- 235000021552 granulated sugar Nutrition 0.000 claims description 10
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 8
- 238000007873 sieving Methods 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 7
- 235000015278 beef Nutrition 0.000 claims description 7
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 7
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 7
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 7
- 239000000284 extract Substances 0.000 claims description 7
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 7
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 7
- CDUFCUKTJFSWPL-UHFFFAOYSA-L manganese(II) sulfate tetrahydrate Chemical compound O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O CDUFCUKTJFSWPL-UHFFFAOYSA-L 0.000 claims description 7
- 239000001632 sodium acetate Substances 0.000 claims description 7
- 235000017281 sodium acetate Nutrition 0.000 claims description 7
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 7
- 239000001393 triammonium citrate Substances 0.000 claims description 7
- 235000011046 triammonium citrate Nutrition 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 6
- 239000007222 ypd medium Substances 0.000 claims description 6
- 238000007605 air drying Methods 0.000 claims description 4
- 239000006872 mrs medium Substances 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- 238000009423 ventilation Methods 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 239000008213 purified water Substances 0.000 claims description 2
- 238000005259 measurement Methods 0.000 claims 1
- 238000005406 washing Methods 0.000 claims 1
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 abstract description 30
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 abstract description 15
- 229960003692 gamma aminobutyric acid Drugs 0.000 abstract description 15
- LIEMBEWXEZJEEZ-INEUFUBQSA-N (2r,3r)-4-(6-aminopurin-9-yl)-2,3-dihydroxybutanoic acid Chemical compound NC1=NC=NC2=C1N=CN2C[C@@H](O)[C@@H](O)C(O)=O LIEMBEWXEZJEEZ-INEUFUBQSA-N 0.000 abstract description 14
- LIEMBEWXEZJEEZ-UHFFFAOYSA-N D-threo-Leutysin Natural products NC1=NC=NC2=C1N=CN2CC(O)C(O)C(O)=O LIEMBEWXEZJEEZ-UHFFFAOYSA-N 0.000 abstract description 14
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- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 210000000936 intestine Anatomy 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- 210000002784 stomach Anatomy 0.000 abstract description 2
- 241000235342 Saccharomycetes Species 0.000 abstract 1
- 229920001491 Lentinan Polymers 0.000 description 18
- 229940115286 lentinan Drugs 0.000 description 18
- 239000000203 mixture Substances 0.000 description 15
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 12
- 239000002994 raw material Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 229920000136 polysorbate Polymers 0.000 description 5
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- 230000000052 comparative effect Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
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- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides a preparation method of mushroom enzyme, which takes mushroom stems as a fermentation main material, firstly preprocesses the mushroom stems, adds sugar and inorganic salt to prepare a mushroom enzyme culture medium, then adds saccharomycetes and lactic acid bacteria step by step to perform standing fermentation, and finally performs after-ripening fermentation, so that the nutrient components of the mushroom stems are effectively dissolved out, the prepared mushroom enzyme has high concentration of polysaccharide and gamma-aminobutyric acid, and contains active substances such as lentinacin, various free amino acids and the like, thereby improving the functions of the obtained mushroom enzyme such as oxidation resistance and the like, improving the functions of blood pressure, reducing blood fat, calming, inhibiting tumor, improving intestines and stomach, regulating arrhythmia, treating epilepsy and the like, in addition, generating additional flavor components, improving the taste and smell of the mushroom enzyme, not only improving the utilization rate and added value of the mushroom stems, and the nutritional value and the health-care value of the mushroom enzyme are also improved.
Description
Technical Field
The invention relates to the technical field of health foods, and particularly relates to a preparation method of mushroom enzyme.
Background
The ferment is a fermented product containing biological active substances and nutrient components obtained by fermenting animals, plants and edible fungi by using microorganisms under proper fermentation conditions. Researches show that the ferment has various saccharides, amino acids, flavonoids, phenolic substances and various biological enzymes, and has the effects of resisting oxidation, promoting digestion, improving human immunity, dispelling the effects of alcohol and protecting liver, relieving allergy symptoms, regulating blood sugar and blood pressure, tranquilizing and diminishing inflammation, supplementing trace elements or vitamins and the like. Different kinds of plants, animals or fungi in nature contain effective components or nutrients to a certain extent, and the content of active substances is improved by fermentation of specific microorganisms, and other active substances which are not contained in the raw materials can be obtained, so that the required nutrients are provided for human bodies or the uncomfortable symptoms of the human bodies are relieved. Most of the current ferment takes fruits as ferment preparation raw materials, the action of microorganisms in the ferment is not fully considered, and more residual raw materials are developed and utilized to process agricultural and sideline products, the functions or effective components of the raw materials are explored, and the raw materials are utilized to the maximum extent.
The fiber content of the mushroom stems is high, the mushroom stems are not easy to chew and have poor taste if being directly eaten, and the mushroom stems are largely discarded in the processing process of mushrooms. However, related researches show that the nutritional ingredients of the lentinus edodes stems and the lentinus edodes pileus are not poor, and the nutritional ingredients except the cellulose are not very different. Therefore, if the mushroom stems can be used for preparing the mushroom enzyme, the preparation method has very important economic significance.
Disclosure of Invention
In view of the above, the present invention provides a preparation method of mushroom enzyme, which uses mushroom stems as a main fermentation material, and comprises the steps of preprocessing mushroom stems, adding sugar and inorganic salt to prepare a mushroom enzyme culture medium, adding yeast and lactic acid bacteria step by step to perform standing fermentation, and performing after-fermentation.
In order to achieve the purpose, the technical scheme of the invention is realized as follows:
a preparation method of mushroom enzyme comprises the following steps:
1) respectively preparing YPD culture medium and MRS culture medium for preparing yeast and lactobacillus liquid seeds;
2) respectively inoculating activated yeast and solid strains of lactobacillus into the YPD culture medium and the MRS culture medium for culture to obtain yeast liquid seeds and lactobacillus liquid seeds;
3) cleaning Lentinus Edodes stem with drinking water, air drying in shade and ventilation, grinding, soaking, pulping, adding white sugar and inorganic salt, stirring to dissolve, and sterilizing to obtain Lentinus Edodes enzyme culture medium;
4) and inoculating the yeast liquid seeds into the mushroom enzyme culture medium for standing fermentation, inoculating the lactic acid bacteria liquid seeds after the yeast standing fermentation is finished, continuing to perform standing fermentation, performing after-ripening fermentation in a low-temperature environment after the lactic acid bacteria standing fermentation is finished, and filtering with clean gauze after the after-ripening fermentation is finished to obtain the mushroom enzyme stock solution.
Optionally, the YPD medium for culturing the liquid yeast seed in the step 1) is prepared by the following method:
10g of yeast extract powder, 20g of peptone and 20g of glucose were dissolved in 1L of purified water, followed by autoclaving at 121 ℃ for 20min to obtain a YPD medium for culturing liquid seeds of yeast.
Optionally, the MRS medium for culturing lactic acid bacteria liquid seeds in step 1) is prepared by the following method:
dissolving 10g of beef extract, 10g of yeast extract, 10g of peptone, 20g of glucose, 2g of dipotassium phosphate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1g of Tween-1 and 2g of triammonium citrate into 1L of pure water, and then carrying out autoclaving at 121 ℃ for 20min to obtain the MRS culture medium for culturing the lactobacillus liquid seeds.
Optionally, the culturing process of inoculating the activated yeast and solid strain of lactic acid bacteria into the YPD medium and the MRS medium, respectively, in step 2) comprises:
inoculating the solid strain of the activated yeast into the YPD culture medium, setting the rotating speed of a shaker to be 120rpm, and culturing at 28 ℃ for 24 h;
inoculating the activated solid strain of lactobacillus into the MRS culture medium, setting the rotation speed of a shaking table at 150rpm, and culturing at 37 ℃ for 48 h.
Optionally, the taking of the mushroom stems in the step 3), cleaning with drinking water, placing in a cool and ventilated place for airing, and then performing milling, soaking and pulping comprises:
cleaning Lentinus Edodes stem with drinking water, air drying in shade and ventilation, grinding, and sieving with 40-100 mesh sieve to obtain Lentinus Edodes stem powder;
soaking the powder in drinking water for 30-90min, and pulping for 30-120 s.
Optionally, when the mushroom stem powder is soaked in drinking water, the material-water ratio of the mushroom stem powder to the drinking water is 1: 10-25 m/v.
Optionally, the amount of the white granulated sugar in the step 3) is 6-12% of the amount of drinking water for soaking the lentinus edodes stem powder; the inorganic salt in the step 3) is magnesium sulfate and potassium dihydrogen phosphate; the consumption of the magnesium sulfate is 0.1% of the consumption of the drinking water for soaking the mushroom stem powder, and the consumption of the monopotassium phosphate is 0.3% of the consumption of the drinking water for soaking the mushroom stem powder.
Optionally, the sterilization process in the step 3) comprises: autoclaving at 121 deg.C for 20 min.
Optionally, the fermentation process of standing fermentation of yeast in the step 4) comprises:
inoculating the yeast liquid seeds into the mushroom enzyme culture medium, and standing and fermenting for 5-10 days at 26-30 ℃, wherein the using amount of the yeast liquid seeds is 6-12% of the drinking water amount for soaking the mushroom stem powder;
the fermentation process of standing fermentation of the lactic acid bacteria in the step 4) comprises the following steps:
after the yeast is subjected to standing fermentation, inoculating the lactic acid bacteria liquid seeds, and continuing to perform standing fermentation for 8-12h at 36-40 ℃, wherein the using amount of the lactic acid bacteria liquid seeds is 1-4% of the drinking water amount for soaking the shiitake stem powder;
the fermentation process of the after-ripening fermentation in the step 4) comprises the following steps: after the standing fermentation of the lactobacillus is finished, after-ripening fermentation is carried out for 12-24h at 4 ℃.
The action mechanism of the invention is as follows:
according to the invention, the yeast is added into the mushroom enzyme culture medium, the biological enzyme and the organic acid secreted by the yeast in the fermentation process can gradually degrade mushroom stems, the nutrient components in the mushroom stems are dissolved out, and the yeast secretes yeast polysaccharide and other nutrient substances, so that the polysaccharide content in the mushroom enzyme is increased, and the oxidation resistance is also improved; according to the invention, the lactobacillus is added into the mushroom enzyme culture medium, the biological enzyme and the organic acid secreted by the lactobacillus in the fermentation process can further degrade the mushroom stems, so that the nutrient components of the mushroom stems are dissolved out more, the lactobacillus ferments the mushroom stems, can generate additional flavor components, improves the smell of the mushroom enzyme, can convert glutamic acid in the mushroom stems into gamma-aminobutyric acid, further increases the nutrient value and the health care value of the mushroom enzyme, enhances the functions of the obtained mushroom enzyme such as oxidation resistance and the like, and has the functions of improving blood pressure, reducing blood fat, calming and the like.
Compared with the prior art, the preparation method of the mushroom enzyme provided by the invention has the following advantages:
1. according to the invention, the yeast and the lactic acid bacteria are added into the mushroom enzyme culture medium step by step, so that the nutrient components of the mushroom stems are effectively dissolved out, additional flavor components are generated, the taste and the smell of the mushroom enzyme are improved, the utilization rate and the added value of the mushroom stems are improved, and the nutrient value and the health care value of the mushroom enzyme are also improved.
2. The mushroom ferment prepared by the invention has high contents of polysaccharide and gamma-aminobutyric acid, and also contains active substances such as lentinacin, various free amino acids and the like, so that the functions of improving the oxidation resistance and the like of the mushroom ferment are improved, and the mushroom ferment has the functions of improving blood pressure, reducing blood fat, calming, inhibiting tumors, improving intestines and stomach, regulating arrhythmia, treating epilepsy and the like.
3. The preparation process is safe and pollution-free, high-end expensive production equipment is not needed, the investment cost is greatly reduced, the fermentation period is short, the fermentation process is simple, the operation is easy, the original active substances of the lentinus edodes stems can be effectively preserved, the nutrition value is high, the taste is good, the smell is pleasant, and the prepared enzyme stock solution can be used for developing a series of functional products.
Detailed Description
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
The present invention will be described in detail with reference to examples.
In the following examples, raw materials or reagents used were purchased from biological or chemical companies unless otherwise specified.
In the following examples, the bacterial species used, unless otherwise specified, may be isolated or purchased by themselves.
Example 1
The preparation method of the mushroom enzyme specifically comprises the following steps:
1) dissolving 10g of yeast extract powder, 20g of peptone and 20g of glucose in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a YPD culture medium for culturing yeast liquid seeds, then inoculating activated yeast solid strains into the YPD culture medium, setting the rotating speed of a shaker at 120rpm, and culturing at 28 ℃ for 24h to obtain yeast liquid seeds;
2) dissolving 10g of beef extract, 10g of yeast extract, 10g of peptone, 20g of glucose, 2g of dipotassium phosphate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1g of Tween and 2g of triammonium citrate in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain an MRS culture medium for culturing lactobacillus liquid seeds, then inoculating the activated lactobacillus solid strains into the MRS culture medium, setting the rotating speed of a shaking table to 150rpm, and culturing at 37 ℃ for 48h to obtain lactobacillus liquid seeds;
3) taking mushroom stems, cleaning the mushroom stems with drinking water, placing the mushroom stems in a cool and ventilated place for airing, then grinding the mushroom stems into powder, sieving the powder by a 60-mesh sieve to obtain mushroom stem powder, then adding the drinking water into the mushroom stem powder according to the material-water ratio of 1: 15(m/v) for soaking for 30min, pulping for 60s to prepare mushroom homogenate, and finally adding 8% of white granulated sugar (8% of the white granulated sugar refers to: the amount of the white granulated sugar is 8% of the amount of drinking water for soaking the lentinus edodes stem powder), and 0.1% magnesium sulfate (0.1% magnesium sulfate refers to: the dosage of the magnesium sulfate is 0.1 percent of the drinking water used for soaking the lentinus edodes stem powder, and 0.3 percent of monopotassium phosphate (0.3 percent of monopotassium phosphate refers to: the amount of the potassium dihydrogen phosphate is 0.3% of the amount of the drinking water used for soaking the lentinus edodes stem powder, stirring until the potassium dihydrogen phosphate is dissolved, and carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a lentinus edodes enzyme culture medium;
4) 9% yeast liquid seed (9% yeast liquid seed refers to: the amount of the yeast liquid seeds is 9 percent of the amount of drinking water for soaking the lentinus edodes stem powder, the yeast liquid seeds are inoculated into the lentinus edodes enzyme culture medium, the lentinus edodes enzyme culture medium is kept stand and fermented for 7 days at the temperature of 28 ℃, and then 3 percent of lactic acid bacteria liquid seeds (3 percent of lactic acid bacteria liquid seeds refer to: the using amount of the lactobacillus liquid seeds is 3 percent of the drinking water amount for soaking the lentinus edodes stem powder, standing and fermenting for 10 days at 37 ℃, then, after-ripening and fermenting for 20 hours at 4 ℃, and filtering by using clean gauze to obtain the lentinus edodes enzyme stock solution.
The lentinan concentration, the gamma-aminobutyric acid concentration, the lentinacin concentration and the DPPH clearance rate of the lentinan stock solution obtained in this example were measured.
Tests prove that the lentinan bulk solution obtained in the embodiment has the polysaccharide concentration of 0.9178mg/mL, the gamma-aminobutyric acid concentration of 84.34mg/L, the lentinacin concentration of 17.76mg/mL, and the DPPH clearance rate of 92.74%.
Example 2
The preparation method of the mushroom enzyme specifically comprises the following steps:
1) dissolving 10g of yeast extract powder, 20g of peptone and 20g of glucose in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a YPD culture medium for culturing yeast liquid seeds, then inoculating activated yeast solid strains into the YPD culture medium, setting the rotating speed of a shaker at 120rpm, and culturing at 28 ℃ for 24h to obtain yeast liquid seeds;
2) dissolving 10g of beef extract, 10g of yeast extract, 10g of peptone, 20g of glucose, 2g of dipotassium phosphate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1g of Tween and 2g of triammonium citrate in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain an MRS culture medium for culturing lactobacillus liquid seeds, then inoculating the activated lactobacillus solid strains into the MRS culture medium, setting the rotating speed of a shaking table to 150rpm, and culturing at 37 ℃ for 48h to obtain lactobacillus liquid seeds;
3) taking mushroom stems, cleaning the mushroom stems with drinking water, placing the mushroom stems in a cool and ventilated place for airing, then grinding the mushroom stems into powder, sieving the powder with a 40-mesh sieve to obtain mushroom stem powder, then adding the drinking water into the mushroom stem powder according to a material-water ratio of 1: 10(m/v) for soaking for 30min, pulping the mixture for 60s to prepare mushroom homogenate, finally adding 8 percent of white granulated sugar (which is 8 percent of the drinking water amount for soaking the mushroom stem powder in the same example 1), 0.1 percent of magnesium sulfate (which is 0.1 percent of the drinking water amount for soaking the mushroom stem powder in the same example 1), 0.3 percent of potassium dihydrogen phosphate (which is 0.3 percent of the drinking water amount for soaking the mushroom stem powder in the same example 1) into the mushroom homogenate, stirring the mixture until the mixture is dissolved, and carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a mushroom enzyme culture medium;
4) inoculating 6% yeast liquid seed (6% of the drinking water amount for soaking the lentinus edodes stem powder in the same example 1) into the lentinus edodes enzyme culture medium, standing and fermenting at 28 ℃ for 7 days, then inoculating 1% lactobacillus liquid seed (1% of the drinking water amount for soaking the lentinus edodes stem powder in the same example 1), standing and fermenting at 37 ℃ for 10 days, then, after-ripening and fermenting at 4 ℃ for 20 hours, and filtering with clean gauze to obtain the lentinus edodes enzyme stock solution.
The lentinan concentration, the gamma-aminobutyric acid concentration, the lentinacin concentration and the DPPH clearance rate of the lentinan stock solution obtained in this example were measured.
Tests prove that the lentinan stock solution obtained in the embodiment has the polysaccharide concentration of 0.8829mg/mL, the gamma-aminobutyric acid concentration of 76.59mg/L, the lentinacin concentration of 15.85mg/mL and the DPPH clearance rate of 89.72%.
Example 3
The preparation method of the mushroom enzyme specifically comprises the following steps:
1) dissolving 10g of yeast extract powder, 20g of peptone and 20g of glucose in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a YPD culture medium for culturing yeast liquid seeds, then inoculating activated yeast solid strains into the YPD culture medium, setting the rotating speed of a shaker at 120rpm, and culturing at 28 ℃ for 24h to obtain yeast liquid seeds;
2) dissolving 10g of beef extract, 10g of yeast extract, 10g of peptone, 20g of glucose, 2g of dipotassium phosphate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1g of Tween and 2g of triammonium citrate in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain an MRS culture medium for culturing lactobacillus liquid seeds, then inoculating the activated lactobacillus solid strains into the MRS culture medium, setting the rotating speed of a shaking table to 150rpm, and culturing at 37 ℃ for 48h to obtain lactobacillus liquid seeds;
3) taking mushroom stems, cleaning the mushroom stems with drinking water, placing the mushroom stems in a cool and ventilated place for airing, then grinding the mushroom stems into powder, sieving the powder with a sieve of 100 meshes to obtain mushroom stem powder, then adding the drinking water into the mushroom stem powder according to the material-water ratio of 1: 25(m/v) for soaking for 30min, pulping the mixture for 60s to prepare mushroom homogenate, finally adding 8 percent of white granulated sugar (which is 8 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1), 0.1 percent of magnesium sulfate (which is 0.1 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1) and 0.3 percent of potassium dihydrogen phosphate (which is 0.3 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1) into the mushroom homogenate, stirring the mixture until the mixture is dissolved, and carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a mushroom enzyme culture medium;
4) inoculating 12% yeast liquid seed (12% of the drinking water amount for soaking Lentinus edodes stem powder in the same example 1) into the Lentinus edodes enzyme culture medium, standing and fermenting at 28 deg.C for 7 days, inoculating 4% lactobacillus liquid seed (4% of the drinking water amount for soaking Lentinus edodes stem powder in the same example 1), standing and fermenting at 37 deg.C for 10 days, subsequently, after-ripening and fermenting at 4 deg.C for 20h, and filtering with clean gauze to obtain Lentinus edodes enzyme stock solution.
The lentinan concentration, the gamma-aminobutyric acid concentration, the lentinacin concentration and the DPPH clearance rate of the lentinan stock solution obtained in this example were measured.
Tests prove that the lentinan bulk solution obtained in the embodiment has the polysaccharide concentration of 0.8916mg/mL, the gamma-aminobutyric acid concentration of 80.65mg/L, the lentinacin concentration of 16.09mg/L and the DPPH clearance rate of 88.94%.
Example 4
The preparation method of the mushroom enzyme specifically comprises the following steps:
1) dissolving 10g of yeast extract powder, 20g of peptone and 20g of glucose in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a YPD culture medium for culturing yeast liquid seeds, then inoculating activated yeast solid strains into the YPD culture medium, setting the rotating speed of a shaker at 120rpm, and culturing at 28 ℃ for 24h to obtain yeast liquid seeds;
2) dissolving 10g of beef extract, 10g of yeast extract, 10g of peptone, 20g of glucose, 2g of dipotassium phosphate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1g of Tween and 2g of triammonium citrate in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain an MRS culture medium for culturing lactobacillus liquid seeds, then inoculating the activated lactobacillus solid strains into the MRS culture medium, setting the rotating speed of a shaking table to 150rpm, and culturing at 37 ℃ for 48h to obtain lactobacillus liquid seeds;
3) taking mushroom stems, cleaning the mushroom stems with drinking water, placing the mushroom stems in a cool and ventilated place for airing, then grinding the mushroom stems into powder, sieving the powder with a 40-mesh sieve to obtain mushroom stem powder, then adding the drinking water into the mushroom stem powder according to a material-water ratio of 1: 15(m/v) for soaking for 30min, pulping the mixture for 60s to prepare mushroom homogenate, finally adding 8 percent of white granulated sugar (which is 8 percent of the drinking water amount for soaking the mushroom stem powder in the same example 1), 0.1 percent of magnesium sulfate (which is 0.1 percent of the drinking water amount for soaking the mushroom stem powder in the same example 1), 0.3 percent of potassium dihydrogen phosphate (which is 0.3 percent of the drinking water amount for soaking the mushroom stem powder in the same example 1) into the mushroom homogenate, stirring the mixture until the mixture is dissolved, and carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a mushroom enzyme culture medium;
4) inoculating 9% yeast liquid seed (9% of the drinking water used for soaking the lentinus edodes stem powder in the same example 1) into the lentinus edodes enzyme culture medium, standing and fermenting at 28 ℃ for 7 days, then inoculating 3% lactobacillus liquid seed (3% of the drinking water used for soaking the lentinus edodes stem powder in the same example 1), standing and fermenting at 37 ℃ for 10 days, then, after-ripening and fermenting at 4 ℃ for 20h, and filtering with clean gauze to obtain the lentinus edodes enzyme stock solution.
The lentinan concentration, the gamma-aminobutyric acid concentration, the lentinacin concentration and the DPPH clearance rate of the lentinan stock solution obtained in this example were measured.
Tests prove that the lentinan stock solution obtained in the embodiment has the polysaccharide concentration of 0.9016mg/mL, the gamma-aminobutyric acid concentration of 81.50mg/L, the lentinacin concentration of 17.54mg/L and the DPPH clearance rate of 91.87%.
Example 5
The preparation method of the mushroom enzyme specifically comprises the following steps:
1) dissolving 10g of yeast extract powder, 20g of peptone and 20g of glucose in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a YPD culture medium for culturing yeast liquid seeds, then inoculating activated yeast solid strains into the YPD culture medium, setting the rotating speed of a shaker at 120rpm, and culturing at 28 ℃ for 24h to obtain yeast liquid seeds;
2) dissolving 10g of beef extract, 10g of yeast extract, 10g of peptone, 20g of glucose, 2g of dipotassium phosphate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1g of Tween and 2g of triammonium citrate in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain an MRS culture medium for culturing lactobacillus liquid seeds, then inoculating the activated lactobacillus solid strains into the MRS culture medium, setting the rotating speed of a shaking table to 150rpm, and culturing at 37 ℃ for 48h to obtain lactobacillus liquid seeds;
3) taking mushroom stems, cleaning the mushroom stems with drinking water, placing the mushroom stems in a cool and ventilated place for airing, then grinding the mushroom stems into powder, sieving the powder with a 60-mesh sieve to obtain mushroom stem powder, then adding the drinking water into the mushroom stem powder according to a material-water ratio of 1: 25(m/v) for soaking for 30min, pulping the mixture for 60s to prepare mushroom homogenate, finally adding 8 percent of white granulated sugar (which is 8 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1), 0.1 percent of magnesium sulfate (which is 0.1 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1) and 0.3 percent of potassium dihydrogen phosphate (which is 0.3 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1) into the mushroom homogenate, stirring the mixture until the mixture is dissolved, and carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a mushroom enzyme culture medium;
4) inoculating 12% yeast liquid seed (12% of the drinking water amount for soaking Lentinus edodes stem powder in the same example 1) into the Lentinus edodes enzyme culture medium, standing and fermenting at 28 deg.C for 7 days, inoculating 4% lactobacillus liquid seed (4% of the drinking water amount for soaking Lentinus edodes stem powder in the same example 1), standing and fermenting at 37 deg.C for 10 days, subsequently, after-ripening and fermenting at 4 deg.C for 20h, and filtering with clean gauze to obtain Lentinus edodes enzyme stock solution.
The lentinan concentration, the gamma-aminobutyric acid concentration, the lentinacin concentration and the DPPH clearance rate of the lentinan stock solution obtained in this example were measured.
Tests prove that the lentinan bulk solution obtained in the embodiment has the polysaccharide concentration of 0.9095mg/mL, the gamma-aminobutyric acid concentration of 80.64mg/L, the lentinacin concentration of 16.72mg/L and the DPPH clearance rate of 90.81%.
Comparative example 1
The preparation method of the mushroom enzyme specifically comprises the following steps:
1) dissolving 10g of yeast extract powder, 20g of peptone and 20g of glucose in 1L of pure water, then carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a YPD culture medium for culturing yeast liquid seeds, then inoculating activated yeast solid strains into the YPD culture medium, setting the rotating speed of a shaker at 120rpm, and culturing at 28 ℃ for 24h to obtain yeast liquid seeds;
2) taking mushroom stems, cleaning the mushroom stems with drinking water, placing the mushroom stems in a cool and ventilated place for airing, then grinding the mushroom stems into powder, sieving the powder with a 60-mesh sieve to obtain mushroom stem powder, then adding the drinking water into the mushroom stem powder according to a material-water ratio of 1: 25(m/v) for soaking for 30min, pulping the mixture for 60s to prepare mushroom homogenate, finally adding 8 percent of white granulated sugar (which is 8 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1), 0.1 percent of magnesium sulfate (which is 0.1 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1) and 0.3 percent of potassium dihydrogen phosphate (which is 0.3 percent of the drinking water amount used for soaking the mushroom stem powder in the same example 1) into the mushroom homogenate, stirring the mixture until the mixture is dissolved, and carrying out high-pressure sterilization at 121 ℃ for 20min to obtain a mushroom enzyme culture medium;
3) inoculating 12% yeast liquid seed (12% of the water used for soaking Lentinus Edodes stem powder in the same example 1) into the above Lentinus Edodes enzyme culture medium, standing and fermenting at 28 deg.C for 7 days, and filtering with clean gauze to obtain Lentinus Edodes enzyme stock solution.
The lentinan concentration, the gamma-aminobutyric acid concentration, the lentinacin concentration and the DPPH clearance rate in the lentinan bulk solution obtained in the comparative example were tested.
Tests show that the lentinan original solution obtained in the comparative example has the polysaccharide content of 0.8659mg/mL, the gamma-aminobutyric acid content of 0.29mg/L, the lentinacin content of 16.72mg/L and the DPPH clearance rate of 89.30%.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (9)
1. The preparation method of the mushroom enzyme is characterized by comprising the following steps:
1) respectively preparing YPD culture medium and MRS culture medium for preparing yeast and lactobacillus liquid seeds;
2) respectively inoculating activated yeast and solid strains of lactobacillus into the YPD culture medium and the MRS culture medium for culture to obtain yeast liquid seeds and lactobacillus liquid seeds;
3) cleaning Lentinus Edodes stem with drinking water, air drying in shade and ventilation, grinding, soaking, pulping, adding white sugar and inorganic salt, stirring to dissolve, and sterilizing to obtain Lentinus Edodes enzyme culture medium;
4) and inoculating the yeast liquid seeds into the mushroom enzyme culture medium for standing fermentation, inoculating the lactic acid bacteria liquid seeds after the yeast standing fermentation is finished, continuing to perform standing fermentation, performing after-ripening fermentation in a low-temperature environment after the lactic acid bacteria standing fermentation is finished, and filtering with clean gauze after the after-ripening fermentation is finished to obtain the mushroom enzyme stock solution.
2. The method for preparing mushroom ferment according to claim 1, wherein the YPD medium for culturing yeast liquid seeds in the step 1) is prepared by the following method:
10g of yeast extract powder, 20g of peptone and 20g of glucose were dissolved in 1L of purified water, followed by autoclaving at 121 ℃ for 20min to obtain a YPD medium for culturing liquid seeds of yeast.
3. The preparation method of the mushroom ferment of claim 1, wherein the MRS medium for culturing the liquid seed of the lactic acid bacteria in the step 1) is prepared by the following method:
dissolving 10g of beef extract, 10g of yeast extract, 10g of peptone, 20g of glucose, 2g of dipotassium phosphate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1g of Tween-1 and 2g of triammonium citrate into 1L of pure water, and then carrying out autoclaving at 121 ℃ for 20min to obtain the MRS culture medium for culturing the lactobacillus liquid seeds.
4. The method for preparing mushroom ferment of claim 1, wherein the culturing process of inoculating the activated yeast and solid strains of lactic acid bacteria into the YPD medium and the MRS medium respectively in the step 2) comprises:
inoculating the solid strain of the activated yeast into the YPD culture medium, setting the rotating speed of a shaker to be 120rpm, and culturing at 28 ℃ for 24 h;
inoculating the activated solid strain of lactobacillus into the MRS culture medium, setting the rotation speed of a shaking table at 150rpm, and culturing at 37 ℃ for 48 h.
5. The preparation method of mushroom enzyme according to claim 1, wherein the step 3) of taking mushroom stems, washing the mushroom stems with drinking water, airing the mushroom stems in a cool and ventilated place, and then grinding, soaking and pulping the mushroom stems comprises:
cleaning Lentinus Edodes stem with drinking water, air drying in shade and ventilation, grinding, and sieving with 40-100 mesh sieve to obtain Lentinus Edodes stem powder;
soaking the powder in drinking water for 30-90min, and pulping for 30-120 s.
6. The preparation method of mushroom ferment as claimed in claim 5, wherein when the mushroom stem powder is soaked in drinking water, the ratio of the mushroom stem powder to the drinking water is 1 to (10-25) m/v.
7. The preparation method of mushroom enzyme according to claim 5, wherein the amount of the white granulated sugar in the step 3) is 6-12% of the amount of drinking water for soaking the mushroom stem powder; the inorganic salt in the step 3) is magnesium sulfate and potassium dihydrogen phosphate; the dosage of the magnesium sulfate is 0.1 percent of the drinking water for soaking the mushroom stem powder, and the dosage of the monopotassium phosphate is 0.1 percent of the drinking water for soaking the mushroom stem powderMeasurement of0.3% of.
8. The preparation method of the mushroom ferment of claim 1, wherein the sterilization process in the step 3) comprises: autoclaving at 121 deg.C for 20 min.
9. The preparation method of mushroom ferment of claim 5, wherein the fermentation process of the yeast stationary fermentation in the step 4) comprises:
inoculating the yeast liquid seeds into the mushroom enzyme culture medium, and standing and fermenting for 5-10 days at 26-30 ℃, wherein the using amount of the yeast liquid seeds is 6-12% of the drinking water amount for soaking the mushroom stem powder;
the fermentation process of standing fermentation of the lactic acid bacteria in the step 4) comprises the following steps:
after the yeast is subjected to standing fermentation, inoculating the lactic acid bacteria liquid seeds, and continuing to perform standing fermentation for 8-12h at 36-40 ℃, wherein the using amount of the lactic acid bacteria liquid seeds is 1-4% of the drinking water amount for soaking the shiitake stem powder;
the fermentation process of the after-ripening fermentation in the step 4) comprises the following steps: after the standing fermentation of the lactobacillus is finished, after-ripening fermentation is carried out for 12-24h at 4 ℃.
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| CN117064055A (en) * | 2023-10-13 | 2023-11-17 | 吉林农业科技学院 | Lucid ganoderma compound enzyme and preparation method and application thereof |
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