CN103948023B - The health food of a kind of develop immunitypty and improving water flood and two-step fermentation preparation method thereof - Google Patents
The health food of a kind of develop immunitypty and improving water flood 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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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Mycology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Environmental Sciences (AREA)
- Botany (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses health food and the two-step fermentation preparation method thereof of a kind of develop immunitypty and improving water flood, the method comprises: glossy ganoderma solid fermentation: with the natural constituents containing develop immunitypty function for property of medicine culture matrix, and inoculation Liquid Strain of Ganoderma Lucidum carries out the medicinal fungal substance that the preparation of two-way solid fermentation has develop immunitypty function; Lactic acid bacteria liquid deep layer fermenting; Take medicinal fungal substance as raw material, add dusty yeast after pulverizing, sodium glutamate, water makes fermentation medium, inoculating lactic acid bacterium carries out liquid deep layer fermenting and produces the GABA promoting sleep, and preparation has the zymotic fluid of develop immunitypty and improving water flood function simultaneously; Adopt zymotic fluid, be prepared into the develop immunitypty of different dosage form and the health food of improving water flood.Health food prepared by the present invention has the function of develop immunitypty and improving water flood simultaneously, wide market, and this preparation method stablizes, and is applicable to large-scale production.
Description
Technical field
The present invention relates to biological health food and preparation field thereof, be specifically related to the health food of a kind of develop immunitypty and improving water flood 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 marches 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 improving the elderly becomes social focus.Hypoimmunity and sleep-disorder are the significant problems affecting senior health and fitness and quality of life, and therefore the health food of develop immunitypty and improving water flood has huge market prospects.
Glossy ganoderma taste is sweet, property is put down, and has the effect of strengthening the body resistance to consolidate the constitution, is classified as top grade 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 commercially produced at present is a lot, 16 sections are had according to the national Bureau of Drugs Supervision website data statistics develop immunitypty of approved and the ganoderma health-care food of improving water flood, these products are raw material mainly with lucidum spore powder, glossy ganoderma or Ganodenna Lucidum P.E greatly, there is the problems such as cost is high, the cycle long, forest tree resource consumption is large.With the ganoderma lucidum mycelium of biofermentation for raw material then can address these problems, as patent 200910186521.X " compound fermentation glossy ganoderma capsule and preparation method thereof " makes the compound product with develop immunitypty, improving water flood with fermentation glossy ganoderma bacterium powder and selenium element formula, patent 96109095.2 " Chinese glossy ganoderma health beverage " prepares glossy ganoderma health beverage with ganoderma lucidum liquid submerged fermentation.
Except traditional glossy ganoderma solid fermentation and liquid fermentation, scholar is had to propose glossy ganoderma novel solid fermentation technique-two-way fermentation technique in recent years, namely Chinese medicine etc. is utilized to ferment containing the property of medicine matrix of bioactive ingredients and glossy ganoderma, this process property of traditional Chinese matrix has an impact to Ganoderma lucidum mycelium bulk-growth, lucid ganoderma fungus enzyme system produces the physiologic effect of 1+1>2 to the transformation of property of medicine matrix, the report of existing a large amount of glossy ganoderma and the two-way fermentation of Chinese medicine at present, 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 and attenuation to property of medicine matrix simultaneously.The two-way fermentation of current glossy ganoderma is only limitted to the fermentation of glossy ganoderma and single medicinal material, the two-way fermentation between the multiple medicinal material that there is no glossy ganoderma and have same effect.
Glossy ganoderma, except having develop immunitypty effect, also has effect of improving water flood, but its onset is slow, needs the onset of long-term taking ability.GABA is a kind of important inhibitory neurotransmitter, and modern study proves that it can play the effect promoting sleep fast, is a kind of quick-acting sleeping factor.GABA and lucid ganoderma complex can play the effect of quick-acting and long-acting improving water flood, there is no the report of GABA and lucid ganoderma complex product at present.
GABA prepared by No. 12nd, the Ministry of Public Health 2009 bulletin approval fermented by lactic acid bacteria is new resource food.The lactic acid bacteria security of screening from traditional fermented food is high, 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 Small molecular such as monose, disaccharides carbon source, directly can not utilize starch-containing and cellulosic natural substrates, and glossy ganoderma has degradable starch and cellulosic enzyme system, starchiness, cellulosic can be decomposed into the Small molecular 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 be decomposed further glossy ganoderma mycoplasma and transform, and more easily absorbs and utilizes.
Summary of the invention
For there is no glossy ganoderma and GABA compound product at present, the invention provides a kind of with function intensified glossy ganoderma medicinal fungal substance and GABA formula, there is the health food of develop immunitypty and improving water flood 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 for the health food of develop immunitypty and improving water flood, comprises the steps:
(1) glossy ganoderma solid fermentation: with the natural constituents containing develop immunitypty function for property of medicine culture matrix, inoculation Liquid Strain of Ganoderma Lucidum carries out the medicinal fungal substance that the preparation of two-way solid fermentation has develop immunitypty function;
(2) lactic acid bacteria liquid deep layer fermenting; With the medicinal fungal substance of step (1) for raw material, add dusty yeast after pulverizing, sodium glutamate, water makes fermentation medium, inoculating lactic acid bacterium carries out liquid deep layer fermenting and produces the GABA promoting sleep, and preparation has the zymotic fluid of develop immunitypty and improving water flood function simultaneously;
(3) adopt the zymotic fluid of step (2), be prepared into the develop immunitypty of different dosage form and the health food of improving water flood.
The first step of the present invention is using the natural constituents containing develop immunitypty function as property of medicine culture matrix, inoculation glossy ganoderma carries out two-way solid fermentation, by property of medicine culture matrix, the impact of ganoderma lucidum mycelium growth metabolism process and lucid ganoderma fungus enzyme system are transformed after interior amphicheirality's solid fermentation property of medicine culture matrix, obtain the medicinal fungal substance having property of medicine matrix and glossy ganoderma develop immunitypty function concurrently; Second step is to contain the medicinal fungal substance of the nutriments such as Small molecular carbon source for raw material after glossy ganoderma degraded, add dusty yeast and sodium glutamate making fluid nutrient medium, the lactic acid bacteria that inoculation can produce GABA is fermented, and preparation has the zymotic fluid of develop immunitypty and improving water flood function; Finally, zymotic fluid through filtering, concentrated, spraying dry, granulation, dress capsule, the technique such as compressing tablet is 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 obtain, and lucidum strain is glossy ganoderma ACCC50621, i.e. the bacterial classification of Chinese agriculture Microbiological Culture Collection administrative center (ACCC) preservation.
In step (1), the inoculum concentration of described Liquid Strain of Ganoderma Lucidum is 2%-10%, the 2%-10%(mL/g that the volume (mL) added of namely described Liquid Strain of Ganoderma Lucidum is the quality (g) of property of medicine culture matrix), with 100g property of medicine culture matrix for 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 first taking 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%, by each component mixing, pulverize, then adding water to moisture is 60%-65%, 116 DEG C-126 DEG C sterilizing 0.5h-1.5h.Preferably, the preparation of described property of medicine culture matrix: the raw material first taking 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%; By each component mixing, pulverize, 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 screened from Pickles, Sichuan Style is: CGMCCNO.5655), namely adopt the bacterial classification of China General Microbiological culture presevation administrative center CGMCC preservation, plant lactobacillus liquid spawn adopts MRS culture medium 28 DEG C of quiescent culture 48h to obtain.
In step (2), described fermentation medium adopts the raw material of following weight portion: water 1000 weight portion, 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, interpolation acetic acid and sodium acetate are mixed with the buffer solution that pH is 5-6,116 DEG C-126 DEG C sterilizing 20min-40min.As preferably, described fermentation medium adopts the raw material of following weight portion: water 1000 weight portion, medicinal fungal substance 50 weight portion, dusty yeast 7.5 weight portion, sodium glutamate 20 weight portion, by water-soluble to medicinal fungal substance, dusty yeast and sodium glutamate, interpolation acetic acid and sodium acetate are mixed with the buffer solution that pH is 5.5,121 DEG C of sterilizing 30min.
In step (2), the inoculum concentration of described lactic acid bacteria is 1%-5%, and the volume added of namely 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, concentrated, spraying dry, granulation, dress capsule, the technique such as compressing tablet is prepared into the health food of different dosage form.
The formulation of described develop immunitypty and the health food of improving water flood is peroral dosage form, be preferably tablet or capsule, fermentation liquor is filtered, concentrated, spraying dry, granulation, obtain tablet after compressing tablet, and fermentation liquor is filtered, concentrated, spraying dry, granulation, obtain capsule after dress capsule.Particularly; first zymotic fluid filters, concentrates, is spray dried to powder; dried particles is obtained through boiling granulating after powder sieves; particle particle pelletizing machine carries out whole grain with 20 mesh sieves; whole grain gained particle adds weight 0.5% disintegrator carboxymethylstarch sodium relative to whole grain gained particle and weight 0.1% magnesium stearate lubricant relative to whole grain gained particle fully mixes 5 minutes encapsulating capsules or is pressed into tablet, every capsules/tablet 0.35g in multi-dimensional movement mixer.
The develop immunitypty prepared by above-mentioned two-step fermentation preparation method and the health food of improving water flood, have the health-care efficacy of develop immunitypty and improving water flood simultaneously.
The two-way fermentation of property of medicine culture matrix of glossy ganoderma and develop immunitypty can prepare the significant medicinal fungal substance of develop immunitypty function, and property of medicine culture matrix is by the degradation of glossy ganoderma, can be lactobacillus-fermented product GABA and 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 adopting glossy ganoderma and lactic acid bacteria two-step fermentation to prepare can the health food of develop immunitypty and improving water flood simultaneously.
Compared with prior art, the present invention has following beneficial effect:
(1) the present invention has develop immunitypty and improving water flood function according to glossy ganoderma, GABA has quick improving water flood function, the two compatibility has the function of remarkable develop immunitypty and improving water flood, provide a kind of with function intensified glossy ganoderma medicinal fungal substance and GABA compatibility can the health food and preparation method thereof of develop immunitypty and improving water flood simultaneously, the method adopts two-step fermenting to organically combine edible fungus fermented with lactobacillus-fermented, wherein the first step utilizes glossy ganoderma to have the medicinal fungal substance of glossy ganoderma and property of medicine matrix enhancement immunity function concurrently with the two-way solid fermentation preparation possessing develop immunitypty function property of medicine culture matrix, 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 the Small molecular carbon source that lactic acid bacteria is easy to utilize, the GABA producing improving water flood for second step lactic acid bacteria liquid deep layer fermenting provides necessary nutrition and carbon source, simultaneously by the degraded of lactic acid bacteria with transform medicinal fungal substance and also more easily absorb, utilize.
(2) health food that prepared by the present invention the experiment proved that to have develop immunitypty and improving water flood function.
(3) preparation method of the present invention's employing, not only cost of material is low, and whole production process all adopts cold fermentation, mild condition, does not have the absorption of harmful toxic matter, and Product Safety, added value are high, can accomplish scale production, wide market.
(4) health food that prepared by the present invention adopts tablet, capsule formulation, is convenient for carrying and takes.
Accompanying drawing explanation
Fig. 1 is the two-step fermentation preparation method flow chart of develop immunitypty of the present invention and improving water flood 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.When being below not particularly illustrated, 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: by formula rate, each component mixed, pulverize, 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 (common process preparation) of glossy ganoderma ACCC50621 is inoculated into 1. property of traditional Chinese culture matrix by 3% volume mass ratio, the i.e. volume (mL) added of the glossy ganoderma ACCC50621 liquid spawn 3%(mL/g that is the quality (g) of property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 25 days has develop immunitypty function for 25 DEG C;
2, lactic acid bacteria liquid deep layer fermenting:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 40g, dusty yeast 5g, sodium glutamate 15g, running water 1L(and 1000g);
Compound method: by formula rate by water-soluble to medicinal fungal substance, dusty yeast, sodium glutamate, adds acetic acid and sodium acetate is mixed with the buffer solution that pH is 5.5, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screened from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (MRS culture medium 28 DEG C of quiescent culture 48h) is inoculated into fermentation medium 1. in 1% ratio, namely the volume added of Lactobacillus plantarum is the zymotic fluid that 1%, 28 DEG C of quiescent culture 72h preparation of fermentation medium volume has develop immunitypty and improving water flood function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; dried particles is obtained through boiling granulating after powder sieves; particle particle pelletizing machine carries out whole grain with 20 mesh sieves; whole grain gained particle adds 0.5% disintegrator carboxymethylstarch sodium relative to whole grain gained particle weight and the abundant mixing in multi-dimensional movement mixer of 0.1% magnesium stearate lubricant relative to whole grain gained particle weight is pressed into tablet in 5 minutes, every sheet 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: by formula rate, each component mixed, pulverize, 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 (common process preparation) of glossy ganoderma ACCC50621 is inoculated into 1. property of traditional Chinese culture matrix by 3% volume mass ratio, the i.e. volume (mL) added of the glossy ganoderma ACCC50621 liquid spawn 3%(mL/g that is the quality (g) of property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 25 days has develop immunitypty function for 25 DEG C;
2, lactic acid bacteria liquid deep layer fermenting:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 40g, dusty yeast 5g, sodium glutamate 15g, running water 1L(and 1000g);
Compound method: by formula rate by water-soluble to medicinal fungal substance, dusty yeast, sodium glutamate, adds acetic acid and sodium acetate is mixed with the buffer solution that pH is 5.5, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screened from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (MRS culture medium 28 DEG C of quiescent culture 48h) is inoculated into fermentation medium 1. in 1% ratio, namely the volume added of Lactobacillus plantarum is the zymotic fluid that 1%, 28 DEG C of quiescent culture 72h preparation of fermentation medium volume has develop immunitypty and improving water flood function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; dried particles is obtained through boiling granulating after powder sieves; particle particle pelletizing machine carries out whole grain with 20 mesh sieves; whole grain gained particle adds 0.5% disintegrator carboxymethylstarch sodium relative to whole grain gained particle weight and the abundant mixing in multi-dimensional movement mixer of 0.1% magnesium stearate lubricant relative to whole grain gained particle weight dresses up capsule in 5 minutes, 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: by formula rate, each component mixed, pulverize, 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 (common process preparation) of glossy ganoderma ACCC50621 is inoculated into 1. property of traditional Chinese culture matrix by 5% volume mass ratio, the i.e. volume (mL) added of the glossy ganoderma ACCC50621 liquid spawn 5%(mL/g that is the quality (g) of property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 20 days has develop immunitypty function for 28 DEG C;
2, lactic acid bacteria liquid deep layer fermenting:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 50g, dusty yeast 7.5g, sodium glutamate 20g, running water 1L(and 1000g);
Compound method: by formula rate by water-soluble to medicinal fungal substance, dusty yeast, sodium glutamate, adds acetic acid and sodium acetate is mixed with the buffer solution that pH is 5.5, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screened from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (MRS culture medium 28 DEG C of quiescent culture 48h) is inoculated into fermentation medium 1. in 1% ratio, namely the volume added of Lactobacillus plantarum is the zymotic fluid that 1%, 32 DEG C of quiescent culture 48h preparation of fermentation medium volume has develop immunitypty and improving water flood function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; dried particles is obtained through boiling granulating after powder sieves; particle particle pelletizing machine carries out whole grain with 20 mesh sieves; whole grain gained particle adds 0.5% disintegrator carboxymethylstarch sodium relative to whole grain gained particle weight and the abundant mixing in multi-dimensional movement mixer of 0.1% magnesium stearate lubricant relative to whole grain gained particle weight is pressed into tablet in 5 minutes, every sheet 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: by formula rate, each component mixed, pulverize, 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 (common process preparation) of glossy ganoderma ACCC50621 is inoculated into 1. property of traditional Chinese culture matrix by 5% volume mass ratio, the i.e. volume (mL) added of the glossy ganoderma ACCC50621 liquid spawn 5%(mL/g that is the quality (g) of property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 20 days has develop immunitypty function for 28 DEG C;
2, lactic acid bacteria liquid deep layer fermenting:
1. fermented and cultured matrix manufacturing
Formula: medicinal fungal substance 50g, dusty yeast 7.5g, sodium glutamate 20g, running water 1L(and 1000g);
Compound method: by formula rate by water-soluble to medicinal fungal substance, dusty yeast, sodium glutamate, adds acetic acid and sodium acetate is mixed with the buffer solution that pH is 5.5, and 121 DEG C of sterilizing 30min, obtain fermentation medium;
2. inoculation fermentation
By the Lactobacillus plantarum LW106(deposit number screened from Pickles, Sichuan Style be: CGMCCNO.5655) liquid spawn (MRS culture medium 28 DEG C of quiescent culture 48h) is inoculated into fermentation medium 1. in 1% ratio, namely the volume added of Lactobacillus plantarum is the zymotic fluid that 1%, 32 DEG C of quiescent culture 48h preparation of fermentation medium volume has develop immunitypty and improving water flood function;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; dried particles is obtained through boiling granulating after powder sieves; particle particle pelletizing machine carries out whole grain with 20 mesh sieves; whole grain gained particle adds 0.5% disintegrator carboxymethylstarch sodium relative to whole grain gained particle weight and the abundant mixing in multi-dimensional movement mixer of 0.1% magnesium stearate lubricant relative to whole grain gained particle weight dresses up capsule in 5 minutes, every 0.35g.
Test example 1
Toxicological test:
Test objective: whether inspection product has toxicity
Test material: the capsule that embodiment 4 is obtained.
Sample: human oral's recommended intake 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: cleaning grade ICR mouse, SD rat and feed.
Rearing conditions: temperature 20-25 DEG C, the barrier system of relative humidity 40%-70%.
Test method:
Chmice acute Oral toxicity is tested: the capsule 's content that Example 4 is obtained, is mixed with 0.500g/mL with sterilized water, and 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: the capsule 's content that Example 4 is obtained, 0.500g/mL is mixed with sterilized water, for the dosage of 5000ug/ ware, 5 dosage are established in test, 1000,200,40,8ug/ ware 4 dosage sterilized waters are mixed with 10 by certain multiple proportions stepwise dilution, 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 endoxan of 40mg/kgbw dosage for positive control, sterilized water is negative control, test group 3 dosage are respectively 10.0g/kgbw, 5.0g/kgbw, 2.5g/kgbw, the capsule 's content sterilized water that Example 4 obtains is 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, second time puts to death animal after giving sample 6h, get the conventional film-making of breastbone, microscopy, every mouse counts 1000 Polychromatic erythrocyte, observe the polychromatic erythrocyte containing micronucleus and observe microkernel incidence.
Mouse inbred strain: with the endoxan of 40mg/kgbw dosage for positive control, sterilized water is negative control, test group 3 dosage are respectively 10.0g/kgbw, 5.0g/kgbw, 2.5g/kgbw, the obtained capsule 's content sterilized water of Example 4 is mixed with 0.5,0.25,0.125g/mL, with 20mL/kgbw gavage, every day 1 time, continuous 5d.Within the 35th day after giving sample first, put to death animal, get the film-making routinely of both sides epididymal sperm filtrate, microscopy, every mouse counts 1000 complete sperms, calculates Sperm malformation rate.
30 days feeding trials: capsules in healthy volunteers recommended amounts 1.4g/ people/sky that embodiment 4 is obtained, 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 in basal feed rat of feeding, continuous nursing 30 days, claim 1 body weight and 2 food intake doses weekly, calculate weekly and total food utilization, every day observes with or without 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 inbred strain result are feminine gender.
3,30 days feeding trial results: experimental session, animal growth is good, each dosage group body weight, food utilization, organ weights, organ coefficient change all without exception, hematological indices and biochemical indices are all in normal range (NR), and each internal organs pathological examination is showed no does not observe illeffects to by the relevant pathological change of test product, the tested material that gives human body recommended amounts 100 times.
Test example 2
Develop immunitypty function test:
Test objective: whether inspection product has the function of develop immunitypty
Test material: the capsule that embodiment 4 is obtained.
Sample: human oral's recommended intake 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: cleaning grade ICR mouse and feed.
Rearing conditions: 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 technical specification " (2003 editions) evaluation criterion, if basic, normal, high 3 dosage and negative control group (sterilized water), basic, normal, high 3 dosage component are not equivalent to people with 5 times, 10 times, 30 times of maximum recommended dosage, i.e. 0.15g/kgbw/d, 0.30g/kgbw/d, 0.90g/kgbw/d, basic, normal, high 3 dosage formulation concentration 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 continuous gavage measures each develop immunitypty functional parameter after one month.
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 of develop immunitypty.
Specific experiment result data is as follows:
Table 1 is on the impact (n=10, x ± s) of the multiplication capacity of the mouse spleen lymphocyte that ConA induces
| Group | Optical density difference DELTA 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
Table 2 is on the impact (n=10, x ± s) of mouse antibodies cellulation
| 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
Table 3 is on the impact (n=10, x ± s) of NK cells in mice activity
| 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
Improving water flood function test:
Test objective: whether inspection product has the function of improving water flood
Test material: the capsule that embodiment 4 is obtained.
Human oral's recommended intake is every day twice, each two, and every 0.35g/ grain (body weight is by 60kg), 0.03g/kgbw, gets capsule 's content and test.
Animal used as test: cleaning grade ICR mouse and feed.
Rearing conditions: 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 technical specification " (2003 editions) evaluation criterion, extend the barbital sodium experiment length of one's sleep, the hypnosis of barbital sodium sub-threshold dose is tested, barbital sodium Sleep Latency Test, if low, in, high 3 dosage and negative control group (sterilized water), low, in, high 3 dosage component are not equivalent to people with 5 times of maximum recommended dosage, 10 times, 30 times, i.e. 0.15g/kgbw/d, 0.30g/kgbw/d, 0.90g/kgbw/d, low, in, high 3 dosage formulation concentration 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 after continuous 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 obviously can shorten barbital sodium induced hypnotic (P<0.05 in incubation period, 0.01, 0.01), equal incidence (the P<0.05 that can significantly improve the sleep of sub-threshold dose barbital sodium inducing mouse, 0.05, 0.01), middle high dose group obviously can extend (the P<0.05 length of one's sleep of barbital sodium induction, 0.01), illustrate that the obtained capsule of embodiment 4 is improved the effect of sleep, and improving water flood act as dose-effect relationship.Specific experiment result data is as follows:
Table 4 is on the impact (n=15, x ± s) of the barbital sodium inducing mouse length of one's sleep
| 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
Table 5 is on the impact (n=15, x ± s) of sub-threshold dose barbital sodium inducing mouse sleep incidence
| 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
Table 6 is on the impact (n=15, x ± s) of barbital sodium inducing mouse Sleep latency
| 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 (9)
1. a two-step fermentation preparation method for the health food of develop immunitypty and improving water flood, is characterized in that, comprise the steps:
(1) glossy ganoderma solid fermentation: with the natural constituents containing develop immunitypty function for property of medicine culture matrix, inoculation Liquid Strain of Ganoderma Lucidum carries out the medicinal fungal substance that the preparation of two-way solid fermentation has develop immunitypty function;
The preparation of described property of medicine culture matrix: the raw material first taking 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%, by each component mixing, pulverize, then adding water to moisture is 60%-65%, 116 DEG C-126 DEG C sterilizing 0.5h-1.5h;
(2) lactic acid bacteria liquid deep layer fermenting; With the medicinal fungal substance of step (1) for raw material, add dusty yeast after pulverizing, sodium glutamate, water makes fermentation medium, inoculating lactic acid bacterium carries out liquid deep layer fermenting and produces the GABA promoting sleep, and preparation has the zymotic fluid of develop immunitypty and improving water flood function simultaneously;
(3) adopt the zymotic fluid of step (2), be prepared into the develop immunitypty of different dosage form and the health food of improving water flood.
2. the two-step fermentation preparation method of the health food of develop immunitypty according to claim 1 and improving water flood, 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 develop immunitypty according to claim 1 and improving water flood, it is characterized in that, in step (1), the preparation of described property of medicine culture matrix: the raw material first taking 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%; By each component mixing, pulverize, then adding water to moisture is 60%-65%, 121 DEG C of sterilizing 1h.
4. the two-step fermentation preparation method of the health food of develop immunitypty according to claim 1 and improving water flood, it 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.
5. the two-step fermentation preparation method of the health food of develop immunitypty according to claim 1 and improving water flood, it is characterized in that, in step (2), described fermentation medium adopts the raw material of following weight portion: water 1000 weight portion, 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, interpolation acetic acid and sodium acetate are mixed with the buffer solution that pH is 5-6,116 DEG C-126 DEG C sterilizing 20min-40min.
6. the two-step fermentation preparation method of the health food of develop immunitypty according to claim 1 and improving water flood, is characterized in that, in step (2), the inoculum concentration of described lactic acid bacteria is 1%-5%.
7. the two-step fermentation preparation method of the health food of develop immunitypty according to claim 1 and improving water flood, it 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.
8. the two-step fermentation preparation method of the health food of develop immunitypty according to claim 1 and improving water flood, is characterized in that, in step (3), described develop immunitypty and the health food of improving water flood are tablet or capsule.
9. the health food of develop immunitypty prepared by the preparation method according to any one of claim 1 ~ 8 and improving water flood.
Priority Applications (1)
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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 |
| CN105145120B (en) * | 2015-09-29 | 2017-11-07 | 古田县聚珍堂食品有限公司 | A kind of cultivating white fungus method |
| CN106235338A (en) * | 2016-08-23 | 2016-12-21 | 广西创瑞生物科技开发有限公司 | A kind of enzyme nutrient solution and preparation method thereof |
| 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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| CN1511193A (en) * | 2001-05-07 | 2004-07-07 | ¹��������ʽ���� | Method for preparing lactic acid fermented solution of mushroom and lactic acid fermented solution of mushroom produced thereby |
| CN103316058A (en) * | 2013-05-22 | 2013-09-25 | 甘肃农业大学 | Technology for improving drug effects of Mythic Fungus and Chinese angelica through utilizing Mythic Fungus-Chinese angelica bidirectional fermentation |
| CN103540624A (en) * | 2012-07-17 | 2014-01-29 | 蔡国珍 | Method for preparing gamma-aminobutyric acid through fermenting cordyceps fungus in rice medium and application of method |
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| JP2004277391A (en) * | 2003-03-14 | 2004-10-07 | Internatl Scient:Kk | Treatment of cancer, diabetes, leukemia and insomnia with sake brewed using deuterium-reduced water and its use for rejuvenation, activation of cells, and acceleration of increase of immunocyte, and production method thereof |
| KR101247935B1 (en) * | 2010-10-15 | 2013-03-29 | 농업회사법인 제주홍암가 주식회사 | Method for Preparing Raw Material for Functional Food |
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| CN1511193A (en) * | 2001-05-07 | 2004-07-07 | ¹��������ʽ���� | Method for preparing lactic acid fermented solution of mushroom and lactic acid fermented solution of mushroom produced thereby |
| CN103540624A (en) * | 2012-07-17 | 2014-01-29 | 蔡国珍 | Method for preparing gamma-aminobutyric acid through fermenting cordyceps fungus in rice medium and application of method |
| CN103316058A (en) * | 2013-05-22 | 2013-09-25 | 甘肃农业大学 | Technology for improving drug effects of Mythic Fungus and Chinese angelica through utilizing Mythic Fungus-Chinese angelica bidirectional fermentation |
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