CN103948023A - Health foods for enhancing immunity and improving sleep and two-step fermentation preparation method thereof - Google Patents
Health foods for enhancing immunity and improving sleep and two-step fermentation preparation method thereof Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- 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
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- 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
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- 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
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- Food Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
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- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
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Abstract
The invention discloses health foods for enhancing immunity and improving sleep and a two-step fermentation preparation method thereof. The method includes the steps: carrying out ganoderma lucidum solid fermentation comprising the steps of taking a natural component having an immunity enhancing function as a medicinal culture medium, inoculating with a ganoderma lucidum liquid strain, carrying out bi-directional solid fermentation, and preparing a medicinal mycoplasm having the immunity enhancing function; carrying out lactic acid bacteria liquid submerged fermentation comprising the steps of taking the medicinal mycoplasm as a raw material, smashing, then adding a yeast powder, monosodium glutamate and water to obtain a fermentation culture medium, inoculating with lactic acid bacteria, carrying out liquid submerged fermentation to produce gamma-aminobutyric acid capable of promoting sleep, and preparing a fermentation liquid simultaneously having an immunity enhancing function and a sleep improving function; and adopting the fermentation liquid, and preparing the health foods for enhancing immunity and improving sleep and with different dosage forms. The prepared health foods simultaneously have the immunity enhancing function and the sleep improving function, and have broad market prospects; and the preparation method is stable, and is suitable for large-scale production.
Description
Technical field
The present invention relates to biological health food and preparation field thereof, be specifically related to a kind ofly strengthen immunity and improve the health food of sleep and adopt glossy ganoderma, lactic acid bacteria solid-liquid two-step fermentation to prepare the method for this health food.
Background technology
Along with the continuous growth of elderly population, China has marched toward aging society, and astogeny process, in continuous acceleration, reaches 2.02 hundred million to the old size of population in 2013 according to statistics, aging level reaches 14.8%, and the Health and Living quality that improves the elderly has become social focus.Hypoimmunity and sleep-disorder are the significant problems that affects senior health and fitness and quality of life, and therefore strengthening immunity and improving the health food of sleeping has huge market prospects.
Glossy ganoderma taste is sweet, property is flat, has the effect of strengthening the body resistance to consolidate the constitution, and is classified as top gradely by Shennong's Herbal, and modern medicine study proves that glossy ganoderma is best immune function controlling agents and activator, can significantly improve the immunologic function of body.The ganoderma lucidum product of having commercially produced is at present a lot, there are 16 sections according to the enhancing immunity of national Bureau of Drugs Supervision website data statistics approved and the ganoderma health-care food of improvement sleep, these products are raw material mainly with lucidum spore powder, glossy ganoderma or Ganodenna Lucidum P.E greatly, have the problems such as cost is high, the cycle long, forest tree resource consumption is large.Can address these problems taking the ganoderma lucidum mycelium of biofermentation as raw material, as patent 200910186521.X " compound fermentation glossy ganoderma capsule and preparation method thereof " makes and has the compound product that strengthens immunity, improves sleep with fermentation glossy ganoderma bacterium powder and selenium element formula, patent 96109095.2 " Chinese glossy ganoderma health beverage " is prepared glossy ganoderma health beverage with ganoderma lucidum liquid submerged fermentation.
Except traditional glossy ganoderma solid fermentation and liquid fermentation, there is in recent years scholar to propose glossy ganoderma novel solid fermentation technique-two-way fermentation technique, utilize Chinese medicine etc. to ferment containing property of medicine matrix and the glossy ganoderma of bioactive ingredients, this process property of traditional Chinese matrix exerts an influence to Ganoderma lucidum mycelium bulk-growth, the physiologic effect of lucid ganoderma fungus enzyme system to property of medicine matrix transformation generation 1+1>2, the report of at present existing a large amount of glossy ganodermas and the two-way fermentation of Chinese medicine, as glossy ganoderma-Radix Astragali, glossy ganoderma-red sage root, glossy ganoderma-rhizome of chuanxiong, glossy ganoderma-radix aconiti agrestis, glossy ganoderma-ginkgo, glossy ganoderma-thunder godvine etc., the two-way fermentation of glossy ganoderma-crotons etc., prove to significantly improve through two-way fermentation glossy ganoderma active component, glossy ganoderma fermentation also has the effect of synergy attenuation to property of medicine matrix simultaneously.The at present two-way fermentation of glossy ganoderma only limits to the fermentation of glossy ganoderma and single medicinal material, there is no glossy ganoderma and has the two-way fermentation between the multiple medicinal material of same effect.
Glossy ganoderma, except having the efficacy of immunity of enhancing, also have effect of improving sleep, but its onset is slow, needs the onset of long-term taking ability.GABA is a kind of important inhibitory neurotransmitter, and it can play the effect that quick promotion is slept modern study proof, is a kind of quick-acting sleeping factor.GABA and lucid ganoderma complex can play the effect of quick-acting and long-acting improvement sleep, there is no at present the report of GABA and lucid ganoderma complex product.
GABA prepared by 2009 No. 12 bulletin approval fermented by lactic acid bacteria of the Ministry of Public Health is new resource food.The lactic acid bacteria of screening from traditional fermented food is safe, can be used for the production of GABA, as the lactic acid bacteria renin Bacillus acidi lactici that patent 200780006225.6 " can produce the lactic acid bacteria of GABA " is separated to from unpurified soy sauce, the Lactobacillus plantarum that patent 200910183478.1 " a strain Efficient Conversion Pidolidone is the seed selection of GABA lactic acid bacteria " is screened from self-control sauerkraut.But lactic acid bacteria can only utilize the little molecule carbon source such as monose, disaccharides, can not directly utilize starch-containing and cellulosic natural substrates, and glossy ganoderma has degradable starch and cellulosic enzyme system, starchiness, cellulosic can be decomposed into the little molecule carbon source that can utilize for lactic acid bacteria, glossy ganoderma fermentation and lactobacillus-fermented are coupled and carry out step fermentation and not only can address this problem, lactic acid bacteria also can further be decomposed and transform glossy ganoderma mycoplasma, more easily absorbs and utilizes.
Summary of the invention
For there is no at present glossy ganoderma and GABA compound product, the invention provides a kind of with function intensified glossy ganoderma medicinal fungal substance and GABA formula, there is the health food that strengthens immunity and improve sleep function simultaneously, provide this health food to utilize the preparation method of glossy ganoderma, lactic acid bacteria two-step fermentation simultaneously.
A two-step fermentation preparation method who strengthens the health food of immunity and improvement sleep, comprises the steps:
(1) glossy ganoderma solid fermentation: to contain the natural constituents that strengthens immunity function as property of medicine culture matrix, inoculation Liquid Strain of Ganoderma Lucidum carries out two-way solid fermentation preparation and has the medicinal fungal substance that strengthens immunity function;
(2) lactic acid bacteria liquid submerged fermentation; Taking the medicinal fungal substance of step (1) as raw material, after pulverizing, interpolation dusty yeast, sodium glutamate, water are made fermentation medium, inoculating lactic acid bacterium carries out liquid deep layer fermenting and produces the GABA that promotes sleep, and preparation has the zymotic fluid that strengthens immunity and improve sleep function simultaneously;
(3) zymotic fluid of employing step (2), is prepared into the enhancing immunity of different dosage form and improves the health food of sleeping.
The first step of the present invention is to contain the natural constituents that strengthens immunity function as property of medicine culture matrix, inoculation glossy ganoderma carries out two-way solid fermentation, impact by property of medicine culture matrix on ganoderma lucidum mycelium growth metabolism process and lucid ganoderma fungus enzyme system transform after interior amphicheirality's solid fermentation property of medicine culture matrix, obtain having concurrently the medicinal fungal substance of property of medicine matrix and glossy ganoderma enhancing immunity function; Second step is taking the medicinal fungal substance that contains the nutriments such as little molecule carbon source after glossy ganoderma degraded as raw material, add dusty yeast and sodium glutamate and make fluid nutrient medium, the lactic acid bacteria that inoculation can produce GABA is fermented, and preparation has the zymotic fluid that strengthens immunity and improve sleep function; The techniques such as finally, zymotic fluid process is filtered, concentrated, spraying is dry, granulation, dress capsule, compressing tablet are prepared into health food.
In step (1), described lucidum strain can adopt prior art, and Liquid Strain of Ganoderma Lucidum adopts existing common process to make, and lucidum strain is glossy ganoderma ACCC50621, i.e. the bacterial classification of Chinese agriculture microorganism fungus kind preservation administrative center (ACCC) preservation.
In step (1), the inoculum concentration of described Liquid Strain of Ganoderma Lucidum is 2%-10%, the volume adding (mL) of described Liquid Strain of Ganoderma Lucidum is the 2%-10%(mL/g of the quality (g) of property of medicine culture matrix), taking 100g property of medicine culture matrix as benchmark, add the Liquid Strain of Ganoderma Lucidum of 2-10mL.
In step (1), the preparation of described property of medicine culture matrix: the raw material that first takes following weight percentage: wholegrain barley 70%-80%, Chinese yam 10%-20%, Radix Astragali 2%-3%, Tiepi Fengdou 1%-2%, American Ginseng 1%-2%, bark of eucommia 1%-2%, barrenwort 1%-2%, land plaster 0.5%-1.5%, each component is mixed, pulverized, then adding water to moisture is 60%-65%, 116 DEG C of-126 DEG C of sterilizing 0.5h-1.5h.Preferably, the preparation of described property of medicine culture matrix: the raw material that first takes following weight percentage: wholegrain barley 75%, Chinese yam 13%, the Radix Astragali 3%, Tiepi Fengdou 2%, American Ginseng 2%, the bark of eucommia 2%, barrenwort 2%, land plaster 1%; Each component is mixed, pulverized, and then adding water to moisture is 60%-65%, 121 DEG C of sterilizing 1h.
In step (1), the condition of described two-way solid fermentation is: fermentation temperature is at 25-28 DEG C, and fermentation time, at 20-25 days, is advisable with this glossy ganoderma solid fermentation condition.
In step (2), described lactic acid bacteria is plant lactobacillus, the Lactobacillus plantarum LW106(deposit number preferably screening from Pickles, Sichuan Style is: CGMCC NO.5655), adopt the bacterial classification of Chinese common micro-organisms culture presevation administrative center CGMCC preservation, plant lactobacillus liquid spawn adopts 28 DEG C of MRS culture mediums to leave standstill and cultivates 48h acquisition.
In step (2), described fermentation medium adopts the raw material of following weight portion: water 1000 weight portions, medicinal fungal substance 40-50 weight portion, dusty yeast 5-10 weight portion, sodium glutamate 15-20 weight portion, by water-soluble to medicinal fungal substance, dusty yeast and sodium glutamate, add acetic acid and sodium acetate and be mixed with the buffer solution that pH is 5-6,116 DEG C of-126 DEG C of sterilizing 20min-40min.As preferably, described fermentation medium adopts the raw material of following weight portion: water 1000 weight portions, medicinal fungal substance 50 weight portions, dusty yeast 7.5 weight portions, sodium glutamate 20 weight portions, by water-soluble to medicinal fungal substance, dusty yeast and sodium glutamate, adding acetic acid and sodium acetate, to be mixed with pH be 5.5 buffer solution, 121 DEG C of sterilizing 30min.
In step (2), the inoculum concentration of described lactic acid bacteria is 1%-5%, and the volume adding of described lactic acid bacteria is the 1%-5% of fermentation medium volume.
In step (2), the condition of described liquid deep layer fermenting is: fermentation temperature is at 28-32 DEG C, and fermentation time, at 48-72h, is advisable with this fermentation condition.
In step (2), in described zymotic fluid, the content of GABA is 9-12g/L.
In step (3), described zymotic fluid, by filtering, the technique such as concentrated, spraying is dry, granulate, dress capsule, compressing tablet is prepared into the health food of different dosage form.
The formulation of the health food of described enhancing immunity and improvement sleep is peroral dosage form, be preferably tablet or capsule, zymotic fluid after filtration, concentrated, spraying is dry, obtain tablet after granulation, compressing tablet, and zymotic fluid after filtration, concentrated, spraying is dry, granulate, obtain capsule after dress capsule.Particularly; first zymotic fluid filters, concentrates, is spray dried to powder; after sieving, powder obtains dried particles through boiling granulating; particle carries out whole grain with particle pelletizing machine with 20 mesh sieves; whole grain gained particle adds with respect to weight 0.5% disintegrator carboxymethylstarch sodium of whole grain gained particle and in multi-dimensional movement mixer, fully mixes 5 minutes encapsulating capsules or be pressed into tablet, every capsules/tablet 0.35g with respect to weight 0.1% magnesium stearate lubricant of whole gained particle.
The health food of the enhancing immunity of preparing by above-mentioned two-step fermentation preparation method and improvement sleep, has the health-care efficacy that strengthens immunity simultaneously and improve sleep.
Glossy ganoderma can be prepared and strengthen the significant medicinal fungal substance of immunity function with the two-way fermentation of property of medicine culture matrix that strengthens immunity, and property of medicine culture matrix is by the degradation of glossy ganoderma, can be lactobacillus-fermented product GABA nutrition and carbon source are provided, simultaneously by the Degradation and Transformation of lactic acid bacteria, property of medicine culture matrix more easily absorbs and utilizes, and therefore adopts glossy ganoderma and lactic acid bacteria two-step fermentation to prepare and can strengthen immunity simultaneously and improve the health food of sleeping.
Compared with prior art, the present invention has following beneficial effect:
(1) the present invention has the immunity of enhancing and improves sleep function according to glossy ganoderma, GABA has the quick sleep function that improves, the two compatibility has remarkable enhancing immunity and improves the function of sleep, a kind of can strengthen immunity simultaneously and improving health food of sleep and preparation method thereof with function intensified glossy ganoderma medicinal fungal substance and GABA compatibility is provided, the method adopts two-step fermenting to organically combine with lactobacillus-fermented edible fungus fermented, wherein the first step utilizes glossy ganoderma and the two-way solid fermentation preparation that possesses enhancing immunity function property of medicine culture matrix to have the medicinal fungal substance of glossy ganoderma and property of medicine matrix enhancement immunity function concurrently, this process can not only produce the physiological function of 1+1 > 2, also the large molecule carbon source in property of medicine culture matrix can be degraded to lactic acid bacteria and be easy to the little molecule carbon source of utilizing, the GABA that produces improvement sleep for the submerged fermentation of second step lactic acid bacteria liquid provides necessary nutrition and carbon source, also more easily absorb by degraded and the conversion medicinal fungal substance of lactic acid bacteria simultaneously, utilize.
(2) health food that prepared by the present invention the experiment proved that to have the immunity of enhancing and improve sleepization function.
(3) preparation method that the present invention adopts, not only cost of material is low, and whole production process all adopts cold fermentation, and mild condition does not have the absorption of harmful toxic matter, and Product Safety, added value are high, can accomplish scale production, and market prospects are wide.
(4) health food that prepared by the present invention adopts tablet, capsule formulation, is convenient for carrying and takes.
Brief description of the drawings
Fig. 1 is that the present invention strengthens immunity and improves the two-step fermentation preparation method flow chart of sleep health food.
Detailed description of the invention
According to Fig. 1 technological process, the present invention is described further below, but protection scope of the present invention is not limited in this.In situation about below not specifying, percentage all refers to percetage by weight.
Embodiment 1
1, glossy ganoderma solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 80%, Chinese yam 13.5%, the Radix Astragali 2%, Tiepi Fengdou 1%, American Ginseng 1%, the bark of eucommia 1%, barrenwort 1%, land plaster 0.5%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 65%; 121 DEG C of sterilizing 1h, obtain property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of glossy ganoderma ACCC50621 (common process preparation) is inoculated into 1. property of traditional Chinese culture matrix by 3% volume mass ratio, the 3%(mL/g of the quality (g) that the volume adding (mL) that is glossy ganoderma ACCC50621 liquid spawn is property of medicine culture matrix), cultivate preparation in 25 days for 25 DEG C and there is the medicinal fungal substance that strengthens immunity function;
2, lactic acid bacteria liquid submerged fermentation:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 40g, dusty yeast 5g, sodium glutamate 15g, running water 1L(is 1000g);
Compound method: press formula rate medicinal fungal substance, dusty yeast, sodium glutamate is water-soluble, adding acetic acid and sodium acetate, to be mixed with pH be 5.5 buffer solution, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (28 DEG C of standing 48h of cultivation of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that 1%, 28 DEG C of fermentation medium volume leaves standstill the zymotic fluid that cultivation 72h preparation has enhancing immunity and improves sleep function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; after sieving, powder obtains dried particles through boiling granulating; particle carries out whole grain with particle pelletizing machine with 20 mesh sieves; whole grain gained particle adds with respect to 0.5% disintegrator carboxymethylstarch sodium of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight and is pressed into tablet, every 0.35g.
Embodiment 2
1, glossy ganoderma solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 80%, Chinese yam 13.5%, the Radix Astragali 2%, Tiepi Fengdou 1%, American Ginseng 1%, the bark of eucommia 1%, barrenwort 1%, land plaster 0.5%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 60%; 121 DEG C of sterilizing 1h, obtain property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of glossy ganoderma ACCC50621 (common process preparation) is inoculated into 1. property of traditional Chinese culture matrix by 3% volume mass ratio, the 3%(mL/g of the quality (g) that the volume adding (mL) that is glossy ganoderma ACCC50621 liquid spawn is property of medicine culture matrix), cultivate preparation in 25 days for 25 DEG C and there is the medicinal fungal substance that strengthens immunity function;
2, lactic acid bacteria liquid submerged fermentation:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 40g, dusty yeast 5g, sodium glutamate 15g, running water 1L(is 1000g);
Compound method: press formula rate medicinal fungal substance, dusty yeast, sodium glutamate is water-soluble, adding acetic acid and sodium acetate, to be mixed with pH be 5.5 buffer solution, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (28 DEG C of standing 48h of cultivation of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that 1%, 28 DEG C of fermentation medium volume leaves standstill the zymotic fluid that cultivation 72h preparation has enhancing immunity and improves sleep function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; after sieving, powder obtains dried particles through boiling granulating; particle carries out whole grain with particle pelletizing machine with 20 mesh sieves; whole grain gained particle adds with respect to 0.5% disintegrator carboxymethylstarch sodium of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight dresses up capsule, every 0.35g.
Embodiment 3
1, glossy ganoderma solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 75%, Chinese yam 13%, the Radix Astragali 3%, Tiepi Fengdou 2%, American Ginseng 2%, the bark of eucommia 2%, barrenwort 2%, land plaster 1%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 63%; 121 DEG C of sterilizing 1h, obtain property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of glossy ganoderma ACCC50621 (common process preparation) is inoculated into 1. property of traditional Chinese culture matrix by 5% volume mass ratio, the 5%(mL/g of the quality (g) that the volume adding (mL) that is glossy ganoderma ACCC50621 liquid spawn is property of medicine culture matrix), cultivate preparation in 20 days for 28 DEG C and there is the medicinal fungal substance that strengthens immunity function;
2, lactic acid bacteria liquid submerged fermentation:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 50g, dusty yeast 7.5g, sodium glutamate 20g, running water 1L(is 1000g);
Compound method: press formula rate medicinal fungal substance, dusty yeast, sodium glutamate is water-soluble, adding acetic acid and sodium acetate, to be mixed with pH be 5.5 buffer solution, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (28 DEG C of standing 48h of cultivation of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that 1%, 32 DEG C of fermentation medium volume leaves standstill the zymotic fluid that cultivation 48h preparation has enhancing immunity and improves sleep function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; after sieving, powder obtains dried particles through boiling granulating; particle carries out whole grain with particle pelletizing machine with 20 mesh sieves; whole grain gained particle adds with respect to 0.5% disintegrator carboxymethylstarch sodium of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight and is pressed into tablet, every 0.35g.
Embodiment 4
1, glossy ganoderma solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 75%, Chinese yam 13%, the Radix Astragali 3%, Tiepi Fengdou 2%, American Ginseng 2%, the bark of eucommia 2%, barrenwort 2%, land plaster 1%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 62%; 121 DEG C of sterilizing 1h, obtain property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of glossy ganoderma ACCC50621 (common process preparation) is inoculated into 1. property of traditional Chinese culture matrix by 5% volume mass ratio, the 5%(mL/g of the quality (g) that the volume adding (mL) that is glossy ganoderma ACCC50621 liquid spawn is property of medicine culture matrix), cultivate preparation in 20 days for 28 DEG C and there is the medicinal fungal substance that strengthens immunity function;
2, lactic acid bacteria liquid submerged fermentation:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 50g, dusty yeast 7.5g, sodium glutamate 20g, running water 1L(is 1000g);
Compound method: press formula rate medicinal fungal substance, dusty yeast, sodium glutamate is water-soluble, adding acetic acid and sodium acetate, to be mixed with pH be 5.5 buffer solution, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (28 DEG C of standing 48h of cultivation of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that 1%, 32 DEG C of fermentation medium volume leaves standstill the zymotic fluid that cultivation 48h preparation has enhancing immunity and improves sleep function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; after sieving, powder obtains dried particles through boiling granulating; particle carries out whole grain with particle pelletizing machine with 20 mesh sieves; whole grain gained particle adds with respect to 0.5% disintegrator carboxymethylstarch sodium of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight dresses up capsule, every 0.35g.
Test example 1
Toxicological test:
Test objective: whether inspection product has toxicity
The capsule that test material: embodiment 4 makes.
Sample: the oral recommended intake of human body is every day twice, each two, every 0.35g/ grain (body weight is by 60kg), 0.03g/kgbw, gets capsule 's content and tests.
Animal used as test: clean level ICR mouse, SD rat and feed.
Raising condition: temperature 20-25 DEG C, the barrier system of relative humidity 40%-70%.
Test method:
Chmice acute Oral toxicity test: get the capsule 's content that embodiment 4 makes, be mixed with 0.500g/mL with sterilized water, by secondary gavage provisions in maximum gavage amount 20mL/kgbw mono-day, it is 20.0g/kgbw that tested material gives dosage.
Salmonella reversion test: get the capsule 's content that embodiment 4 makes, be mixed with 0.500g/mL with sterilized water, for the dosage of 5000ug/ ware, 5 dosage are established in test, 1000,200,40,4 dosage of 8ug/ ware with sterilized water by certain multiple proportions stepwise dilution be mixed with 10,2,0.4,0.08mg/mL, application of sample amount is 0.1mL/ ware, adds 0.1mL/ ware, establish the positive, feminine gender and solvent control simultaneously, count every ware and return change clump count.
Mouse marrow cell micro nuclear test: with the positive contrast of endoxan of 40mg/kgbw dosage, the negative contrast of sterilized water, 3 dosage of test group are respectively 10.0g/kgbw, 5.0g/kgbw, 2.5g/kgbw, get the capsule 's content sterilized water that embodiment 4 makes and be mixed with 0.5, 0.25, 0.125g/mL, with 20mL/kgbw gavage 2 times, adopt 30h to tested material method, give sample interval 24h twice, after giving sample 6h for the second time, put to death animal, get the conventional film-making of breastbone, microscopy, 1000 marrow polychromatic erythrocytes of every mouse counting, observe and contain the polychromatic erythrocyte of micronucleus and observe microkernel incidence.
Mouse sperm deformity test: with the positive contrast of endoxan of 40mg/kgbw dosage, the negative contrast of sterilized water, 3 dosage of test group are respectively 10.0g/kgbw, 5.0g/kgbw, 2.5g/kgbw, get that capsule 's content sterilized water that embodiment 4 makes is mixed with 0.5,0.25,0.125g/mL, with 20mL/kgbw gavage, every day 1 time, continuously 5d.In giving within the 35th day after sample, put to death animal first, get the film-making routinely of both sides epididymal sperm filtrate, microscopy, 1000 complete sperms of every mouse counting, calculate Sperm malformation rate.
Capsules in healthy volunteers recommended amounts 1.4g/ people/sky that 30 days feeding trial: embodiment 4 make, press 30 of human body recommended amounts, 65, establish 0.70g/kgbw/d for 100 times, 1.52g/kgbw/d, 2.33g/kgbw/d establishes three dosage groups, separately establish a blank group 0g/kgbw, given the test agent mixes the rat of feeding in basal feed, feed 30 days continuously, claim weekly 1 body weight and 2 food intake doses, calculate weekly and total food utilization, observe every day and have or not poisoning realization and death, hematology is observed in off-test, blood biochemical, organ coefficient index, and carry out pathological examination.
Result of the test:
1, acute oral toxicity test result: the maximal tolerance dose (MTD) of given the test agent to female male chmice acute per os is all greater than 20.0g/kgbw, belongs to nontoxic level.
2, three genetic toxicity test results: Salmonella reversion test, mouse marrow cell micro nuclear test, mouse sperm deformity result of the test are all negative.
3,30 days feeding trial results: experimental session, animal growth is good, each dosage group body weight, food utilization, organ weights, organ coefficient are all without abnormal change, hematological indices and biochemical indices be all in normal range (NR), and the tested material that each internal organs pathological examination is showed no the pathological change relevant to being subject to test product, give 100 times of human body recommended amounts is not observed illeffects.
Test example 2
Strengthen immunity function test:
Test objective: whether inspection product has the function that strengthens immunity
The capsule that test material: embodiment 4 makes.
Sample: the oral recommended intake of human body is every day twice, each two, every 0.35g/ grain (body weight is by 60kg), 0.03g/kgbw, gets capsule 's content and tests.
Animal used as test: clean level ICR mouse and feed.
Raising condition: temperature 20-25 DEG C, the barrier system of relative humidity 40%-70%.
Test method: carry out zoopery according to " health food inspection and assessment technique specification " (2003 editions) evaluation criterion, if basic, normal, high 3 dosage and negative control group (sterilized water), basic, normal, high 3 dosage groups are equivalent to respectively 5 times, 10 times, 30 times with maximum recommended dosage of people, be 0.15g/kgbw/d, 0.30g/kgbw/d, 0.90g/kgbw/d, basic, normal, high 3 dosage compound concentrations are respectively 15,30,90mg/mL.Per os gives the tested material of mouse corresponding dosage once a day, and mouse stomach amount is 10mL/kgbw, and gavage is measured each enhancing immunity function index after one month continuously.
Experimental result as shown in Table 1, 2 and 3, per os is to mouse 0.15g/kgbw/d, 0.30g/kgbw/d, the given the test agent of basic, normal, high 3 dosage of 0.90g/kgbw/d, middle and high dosage group can significantly improve mouse spleen lymphocyte and transform, improves mouse antibodies cellulation, P<0.05, high dose group can improve NK cells in mice activity, P<0.01.Tested material has no obvious impact (P>0.05) to Mouse Weight, thymus gland/body weight ratio, spleen/body weight ratio, delayed allergy, serum hemolysin and mononuclear macrophage function, illustrates that this sample has the function that strengthens immunity.
Specific experiment result data is as follows:
The impact of the multiplication capacity of the mouse spleen lymphocyte of table 1 on ConA induction (n=10, x ± s)
Group | Optical density difference Δ OD 570nm | P value |
Negative control group | 0.106±0.032 | ? |
Low dose group | 0.112±0.043 | 0.354 |
Middle dosage group | 0.294±0.027 | 0.031* |
High dose group | 0.376±0.038 | 0.019* |
Note: * P<0.05, * * P<0.01
The impact of table 2 on mouse antibodies cellulation (n=10, x ± s)
Group | Hemolysis plaque number (individual/10 6Splenocyte) | P value |
Negative control group | 61±26 | ? |
Low dose group | 67±28 | 0.854 |
Middle dosage group | 89±17 | 0.029* |
High dose group | 92±20 | 0.018* |
Note: * P<0.05, * * P<0.01
The impact of table 3 on NK cells in mice activity (n=10, x ± s)
Group | NK cytoactive (%) | P value |
Negative control group | 31.6±3.9 | ? |
Low dose group | 34.4±8.3 | 0.956 |
Middle dosage group | 27.9±7.5 | 0.735 |
High dose group | 43.9±5.3 | 0.004** |
Note: * P<0.05, * * P<0.01
Test example 3
Improve sleep function test:
Test objective: whether inspection product has the function of improving sleep
The capsule that test material: embodiment 4 makes.
The oral recommended intake of human body is every day twice, each two, and every 0.35g/ grain (body weight is by 60kg), 0.03g/kgbw, gets capsule 's content and tests.
Animal used as test: clean level ICR mouse and feed.
Raising condition: temperature 20-25 DEG C, the barrier system of relative humidity 40%-70%.
Test method: carry out direct hypnosis experiment according to " health food inspection and assessment technique specification " (2003 editions) evaluation criterion, extend the barbital sodium experiment length of one's sleep, barbital sodium sub-threshold dose hypnosis experiment, the test of barbital sodium Sleep latency, if low, in, high 3 dosage and negative control group (sterilized water), low, in, high 3 dosage groups are equivalent to respectively 5 times with maximum recommended dosage of people, 10 times, 30 times, be 0.15g/kgbw/d, 0.30g/kgbw/d, 0.90g/kgbw/d, low, in, high 3 dosage compound concentrations are respectively 15, 30, 90mg/mL.Per os gives the tested material of mouse corresponding dosage once a day, and mouse stomach amount is 10mL/kgbw, measures each sleep index continuously after gavage 30d.
Experimental result is as table 4, shown in table 5 and table 6: per os is to mouse 0.15g/kgbw/d, 0.30g/kgbw/d, 0.90g/kgbw/d is low, in, the given the test agent of high 3 dosage, low, in, high 3 dosage groups all can obviously shorten barbital sodium induced hypnotic (P<0.05 in incubation period, 0.01, 0.01), all can obviously improve the incidence (P<0.05 of sub-threshold dose barbital sodium inducing mouse sleep, 0.05, 0.01), middle high dose group can obviously extend (the P<0.05 length of one's sleep of barbital sodium induction, 0.01), illustrate that capsule that embodiment 4 makes is improved the effect of sleep, and improve the dose-effect relationship that acts as of sleep.Specific experiment result data is as follows:
Table 4 is on the impact of the length of one's sleep of barbital sodium inducing mouse (n=15, x ± s)
Group | The length of one's sleep (min) | P value |
Negative control group | 19.5±9.7 | ? |
Low dose group | 26.8±5.9 | 0.823 |
Middle dosage group | 36.5±8.3 | 0.034* |
High dose group | 45.2±11.3 | 0.007** |
Note: * P<0.05, * * P<0.01
The impact of table 5 on sub-threshold dose barbital sodium inducing mouse sleep incidence (n=15, x ± s)
Group | Sleep incidence (%) | P value |
Negative control group | 20 | ? |
Low dose group | 66.4 | 0.021* |
Middle dosage group | 80 | 0.012* |
High dose group | 90 | 0.003** |
Note: * P<0.05, * * P<0.01
The impact of table 6 on barbital sodium inducing mouse Sleep latency (n=15, x ± s)
Group | The length of one's sleep (min) | P value |
Negative control group | 35.8±10.5 | ? |
Low dose group | 29.3±5.9 | 0.048* |
Middle dosage group | 25.4±8.1 | 0.009** |
High dose group | 20.2±9.3 | 0.006** |
Claims (10)
1. a two-step fermentation preparation method who strengthens the health food of immunity and improvement sleep, is characterized in that, comprises the steps:
(1) glossy ganoderma solid fermentation: to contain the natural constituents that strengthens immunity function as property of medicine culture matrix, inoculation Liquid Strain of Ganoderma Lucidum carries out two-way solid fermentation preparation and has the medicinal fungal substance that strengthens immunity function;
(2) lactic acid bacteria liquid submerged fermentation; Taking the medicinal fungal substance of step (1) as raw material, after pulverizing, interpolation dusty yeast, sodium glutamate, water are made fermentation medium, inoculating lactic acid bacterium carries out liquid deep layer fermenting and produces the GABA that promotes sleep, and preparation has the zymotic fluid that strengthens immunity and improve sleep function simultaneously;
(3) zymotic fluid of employing step (2), is prepared into the enhancing immunity of different dosage form and improves the health food of sleeping.
2. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 1 and improvement sleep, is characterized in that, in step (1), the inoculum concentration of described Liquid Strain of Ganoderma Lucidum is 2%-10%.
3. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 1 and improvement sleep, it is characterized in that, in step (1), the preparation of described property of medicine culture matrix: the raw material that first takes following weight percentage: wholegrain barley 70%-80%, Chinese yam 10%-20%, Radix Astragali 2%-3%, Tiepi Fengdou 1%-2%, American Ginseng 1%-2%, bark of eucommia 1%-2%, barrenwort 1%-2%, land plaster 0.5%-1.5%, each component is mixed, pulverized, then adding water to moisture is 60%-65%, 116 DEG C of-126 DEG C of sterilizing 0.5h-1.5h.
4. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 3 and improvement sleep, it is characterized in that, in step (1), the preparation of described property of medicine culture matrix: the raw material that first takes following weight percentage: wholegrain barley 75%, Chinese yam 13%, the Radix Astragali 3%, Tiepi Fengdou 2%, American Ginseng 2%, the bark of eucommia 2%, barrenwort 2%, land plaster 1%; Each component is mixed, pulverized, and then adding water to moisture is 60%-65%, 121 DEG C of sterilizing 1h.
5. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 1 and improvement sleep, is characterized in that, in step (1), the condition of described two-way solid fermentation is: fermentation temperature is at 25-28 DEG C, and fermentation time is at 20-25 days.
6. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 1 and improvement sleep, it is characterized in that, in step (2), described fermentation medium adopts the raw material of following weight portion: water 1000 weight portions, medicinal fungal substance 40-50 weight portion, dusty yeast 5-10 weight portion, sodium glutamate 15-20 weight portion, by water-soluble to medicinal fungal substance, dusty yeast and sodium glutamate, add acetic acid and sodium acetate and be mixed with the buffer solution that pH is 5-6,116 DEG C of-126 DEG C of sterilizing 20min-40min.
7. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 1 and improvement sleep, is characterized in that, in step (2), the inoculum concentration of described lactic acid bacteria is 1%-5%.
8. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 1 and improvement sleep, is characterized in that, in step (2), the condition of described liquid deep layer fermenting is: fermentation temperature is at 28-32 DEG C, and fermentation time is at 48-72h.
9. the two-step fermentation preparation method of the health food of enhancing immunity according to claim 1 and improvement sleep, is characterized in that, in step (3), the health food of described enhancing immunity and improvement sleep is tablet or capsule.
10. the health food of the enhancing immunity of preparing according to the preparation method described in claim 1~9 any one and improvement sleep.
Priority Applications (1)
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CN110218656A (en) * | 2019-05-31 | 2019-09-10 | 南京工业大学 | The two-way solid fermentation method of ganoderma lucidum-dregs of a decoction and application using air pressure pulsation |
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CN104856172A (en) * | 2015-04-03 | 2015-08-26 | 苏州葛家坞生物科技有限公司 | High gamma-aminobutyric acid content ganoderma lucidum beverage preparation method and product thereof |
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CN110218656A (en) * | 2019-05-31 | 2019-09-10 | 南京工业大学 | The two-way solid fermentation method of ganoderma lucidum-dregs of a decoction and application using air pressure pulsation |
CN110218656B (en) * | 2019-05-31 | 2022-05-17 | 南京工业大学 | Ganoderma lucidum-herb residue bidirectional solid fermentation method utilizing air pressure pulsation and application |
CN112106981A (en) * | 2020-09-21 | 2020-12-22 | 绿之韵生物工程集团有限公司 | Anti-fatigue ganoderma lucidum preparation and preparation method thereof |
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