CN103932181A - Functional food for relieving pressure and improving memory and secondary fermentation preparation method thereof - Google Patents

Functional food for relieving pressure and improving memory and secondary fermentation preparation method thereof Download PDF

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Publication number
CN103932181A
CN103932181A CN201410136197.1A CN201410136197A CN103932181A CN 103932181 A CN103932181 A CN 103932181A CN 201410136197 A CN201410136197 A CN 201410136197A CN 103932181 A CN103932181 A CN 103932181A
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fermentation
functional food
pressure
preparation
memory
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CN103932181B (en
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吴学谦
徐娟
熊科辉
党亚丽
俞盈
许海顺
李青青
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Junliantikang Shandong Pharmaceutical Co ltd
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Zhejiang Academy of Medical Sciences
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L31/00Edible extracts or preparations of fungi; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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  • Mycology (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Environmental Sciences (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention discloses functional food for relieving pressure and improving memory and a secondary fermentation preparation method thereof. The method comprises the following steps: carrying out enoki mushroom solid fermentation, namely, inoculating an enoki mushroom liquid strain by taking a natural component capable of improving memory as a medicinal culture medium, and implementing bidirectional solid fermentation to prepare medicinal mycoplasm having a function of improving memory; carrying out lactobacillus liquid submerged fermentation, namely, grinding the medicinal mycoplasm which serves as a raw material, then adding yeast powder, sodium glutamate and water to prepare a fermentation medium, inoculating lactobacillus and implementing liquid submerged fermentation to produce gamma-aminobutyric acid capable of relieving pressure, thus preparing fermentation liquor simultaneously having functions of relieving pressure and improving memory; preparing functional food for relieving pressure and improving memory in various dosage forms through the fermentation liquor. The prepared functional food disclosed by the invention simultaneously has functions of relieving pressure and improving memory, and is wide in market prospect; the preparation method is stable, and is applicable to large-scale production.

Description

A kind of functional food and two-step fermentation preparation method thereof who releases the pressure and improve memory
Technical field
The present invention relates to functional food and preparation field thereof, be specifically related to a kind ofly release the pressure and improve the functional food of memory and adopt asparagus, lactic acid bacteria solid-liquid two-step fermentation to prepare the method for this functional food.
Background technology
Continuous quickening along with modern life rhythm, people are in life, study, competition in work is more and more fierce, stress is large, the inferior health problems such as failure of memory become puzzlement and comprise student, white collar is in interior mental worker's major hidden danger, therefore increasing people starts to pay close attention to " brain is healthy ", release the pressure, improving food and the health food of memory is more and more favored, the current health food for brain health is to strengthen immunity, antifatigue, it is main improving memory, and be mostly the product of simple function, there is no the health food that can release the pressure simultaneously and improve memory.
Asparagus is described as " increasing intelligence mushroom ", and research shows that flammulina velutipes extract has the effect of improving memory.At present mostly asparagus health food be take asparagus fructification and is prepared as raw material, as mono-kind of patent 201310071534.9 < < has the health food > > gold needle mushroom extract of eyesight-improving intelligence-developing effect, Europe blueberry extract, ginkgo biloba p.e, the compatibilities such as soybean lecithin form, patent 201010622493.4 < < composition > > for alleviating asthenopia and auxiliary improvement of memory is by whortle extract, gold needle mushroom extract, the compatibilities such as taurine form, but use asparagus fructification for existing cost of material high, the problems such as unstable product quality, the golden mushroom mycelium of take can be avoided these problems as raw material, as patent 93110333.9 < < " Jinzhengu " edible fungus beverages and preparation method thereof > > is mainly used golden mushroom mycelium formulated.But golden mushroom mycelium is all with conventional solid or liquid fermentation preparation, there is no the report that utilizes asparagus and the property of medicine matrix with certain effect to prepare by two-way fermentation.
Zhuan Yi etc. propose the concept of two-way fermentation first, point out that edible and medicinal fungi and the Chinese medicine that contains active component ferment and have amphicheirality, can produce the physiologic effect of 1+1>2, as a patent 201110271923.7 < < fungi medicinal fungal substance with effect of lowering blood sugar and preparation method thereof > > selects, there is the property of medicine matrix of effect of lowering blood sugar and fungi strain and obtain and there is obvious reduction blood sugar and do not increase again the medicinal fungal substance of the effect of insulin concentration as two-way fermentation, the Zymocyte mixture that patent 200910058889.8 < < contain glossy ganoderma, preparation method and application > > thereof carry out two-way fermentation with glossy ganoderma and the property of medicine matrix that contains the red sage root, obtain having concurrently the mycoplasma fermentation end-product of red rooted salvia and lucid ganoderma fungus effect.Select asparagus and have and improve the property of medicine matrix of memory effect and carry out solid fermentation, by property of medicine matrix, the impact of golden mushroom mycelium growth metabolism process and asparagus fungal enzyme system are transformed after interior amphicheirality's solid fermentation property of medicine matrix, obtain having concurrently the medicinal fungal substance of property of medicine matrix and asparagus improvement memory effect, can be used as healthy food material, to failure of memory person, can produce the effect of 1+1>2.
GABA is a kind of important inhibitory neurotransmitter, the effect that modern medicine study proof GABA has calm nerve, releases the pressure, 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, the lactic acid bacteria > > that can produce GABA as patent 200780006225.6 < < is separated to the renin Bacillus acidi lactici that can produce GABA (GABA) from unpurified soy sauce, patent 200910183478.1 < < mono-strain Efficient Conversion Pidolidones are that the seed selection > > of GABA lactic acid bacteria filters out the Lactobacillus plantarum that a strain can utilize Pidolidone production GABA from self-control sauerkraut.But lactic acid bacteria can only utilize the little molecule carbon sources such as monose, disaccharides, can not directly utilize starch-containing and cellulosic natural substrates, and fungi 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, fungi fermentation and lactobacillus-fermented are coupled and carry out step fermentation and can address this problem.
Summary of the invention
For there is no at present the functional food of releasing the pressure and improving memory, the invention provides and a kind ofly release the pressure and improve the functional food of memory and utilize asparagus, lactic acid bacteria solid-liquid two-step fermentation to prepare the method for this functional food.
Release the pressure and improve the two-step fermentation preparation method of the functional food of memory, comprising the steps:
(1) asparagus solid fermentation: take containing the natural constituents that is improved memory effect is property of medicine culture matrix, inoculation flammulina velutipes liquid strains carries out two-way solid fermentation preparation and has the medicinal fungal substance that improves memory effect;
(2) lactic acid bacteria liquid submerged fermentation: the medicinal fungal substance of step (1) of take is 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 of releasing the pressure, and preparation has releasing the pressure simultaneously and improves the zymotic fluid of memory effect;
(3) adopt the zymotic fluid of step (2), be prepared into releasing the pressure and improving the functional food of memory of different dosage form.
The first step of the present invention is usingd containing the natural constituents that is improved memory effect as property of medicine culture matrix, inoculation asparagus carries out two-way solid fermentation, by property of medicine culture matrix, the impact of golden mushroom mycelium growth metabolism process and asparagus fungal enzyme system are transformed after interior amphicheirality's solid fermentation property of medicine matrix, obtain having concurrently the medicinal fungal substance of property of medicine matrix and asparagus improvement memory effect; It is raw material that second step be take the medicinal fungal substance that contains the nutriments such as little molecule carbon source after asparagus degraded, add dusty yeast and sodium glutamate and make fermentation medium, the lactic acid bacteria that inoculation can produce GABA is fermented, and preparation has releasing the pressure and improves the zymotic fluid of memory effect; Finally, zymotic fluid through filtering, the technique such as concentrated, preparation prepares health food.
In step (1), described asparagus can adopt prior art, and flammulina velutipes liquid strains adopts existing common process to make, as specifically selected asparagus ACCC50424, i.e. and the bacterial classification of Chinese agriculture microorganism fungus kind preservation administrative center (ACCC) preservation.
In step (1), the inoculum concentration of described flammulina velutipes liquid strains is 2%-10%, the volume adding (mL) of described flammulina velutipes liquid strains is the 2%-10%(mL/g of the quality (g) of property of medicine culture matrix), the 100g property of medicine culture matrix of take is benchmark, adds the flammulina velutipes liquid strains 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%, soybean 10%-20%, walnut kernel 1%-3%, fructus alpiniae oxyphyllae 1%-3%, ginkgo leaf 1%-3%, land plaster 0.5%-1.5%, each component is mixed, pulverized, then adding water to moisture weight content is 60%-65%, 116 ℃ of-126 ℃ 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%, soybean 15%, walnut kernel 3%, fructus alpiniae oxyphyllae 3%, ginkgo leaf 3%, land plaster 1%, each component is mixed, pulverized, then adding water to moisture weight content is 60%-65%, 121 ℃ of sterilizing 1h.
In step (1), the condition of described two-way solid fermentation is: fermentation temperature is at 20-25 ℃, and fermentation time, at 20-30 days, is advisable with this asparagus 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 ℃ of standing cultivation 48h of MRS culture medium to obtain.
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, medicinal fungal substance, dusty yeast and sodium glutamate is water-soluble, add acetic acid and sodium acetate and be mixed with the buffer solution that pH is 5-6,116 ℃ of-126 ℃ 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, medicinal fungal substance, dusty yeast and sodium glutamate is water-soluble, adding acetic acid and sodium acetate, to be mixed with pH be 5.5 buffer solution, 121 ℃ 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 ℃, 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, preparation is prepared into the functional food of different dosage form.
The described formulation of releasing the pressure and improving the functional food of memory 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 filter, concentrate, be spray dried to powder; powder sieves and obtains dried particles by 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 with respect to weight 0.1% magnesium stearate lubricant 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.
That by above-mentioned two-step fermentation preparation method, prepares releases the pressure and improves the functional food of memory, and this functional food has releasing the pressure simultaneously and improves the function of memory.
Asparagus can be prepared and improve the significant medicinal fungal substance of memory effect with the two-way fermentation of property of medicine culture matrix that improves memory, and property of medicine culture matrix is by the degradation of asparagus, medicinal fungal substance can be lactobacillus-fermented product GABA nutrition and carbon source is provided, by degraded and the conversion of lactic acid bacteria, medicinal fungal substance more easily absorbs and utilizes simultaneously.Therefore adopt asparagus and lactic acid bacteria two-step fermentation can prepare the functional food that can release the pressure simultaneously and improve memory.
Compared with prior art, the present invention has following beneficial effect:
(1) the present invention has the memory of improvement effect according to asparagus, GABA has the effect of releasing the pressure, the two compatibility has releasing the pressure and improves memory effect, provide a kind of with function intensified asparagus medicinal fungal substance and the preparation method of functional food of GABA compatibility, the method organically combines with lactobacillus-fermented edible fungus fermented, adopt two-step fermenting preparation can release the pressure simultaneously and improve the functional food of memory, wherein the first step is utilized asparagus and is possessed improvement and remembers the medicinal fungal substance that the two-way solid fermentation preparation of effect property of medicine matrix has asparagus and property of medicine matrix improvement memory effect concurrently, this process can not only produce the physiological function of 1+1 > 2, also the large molecule carbon source in property of medicine matrix can be degraded to lactic acid bacteria and be easy to the little molecule carbon source of utilizing, the GABA that possesses the effect of releasing the pressure for second step lactic acid bacteria liquid submerged fermentation generation provides necessary nutrition and carbon source, simultaneously by degraded and the conversion of lactic acid bacteria, medicinal fungal substance more easily absorbs and utilizes.
(2) functional food that prepared by the present invention the experiment proved that to have releasing the pressure and improve memory function.
(3) production 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) functional food that prepared by the present invention adopts the formulations such as tablet, capsule, is convenient for carrying and takes.
Accompanying drawing explanation
Fig. 1 is that the present invention releases the pressure and improves two-step fermentation preparation method's the flow chart of the functional food of memory;
Fig. 2 is the rate of change (starting point 100%) of experimenter's saliva cortisol in test example 2.
The specific embodiment
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.Below do not have in the situation of special instruction, percentage all refers to percetage by weight.
Embodiment 1
1, asparagus solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 80%, soybean 16.5%, walnut kernel 1%, fructus alpiniae oxyphyllae 1%, ginkgo leaf 1%, land plaster 0.5%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 65%, 121 ℃ of sterilizing 1h, obtains property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of asparagus ACCC50424 (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 asparagus ACCC50424 liquid spawn is property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 30 days has improvement memory effect for 20 ℃;
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 ℃ of sterilizing 30min, obtain fermentation medium;
2. by the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style, be: CGMCCNO.5655) liquid spawn (28 ℃ of standing cultivation 48h of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that the standing cultivation of 1%, 28 ℃ of fermentation medium volume 72h preparation has releasing the pressure and improves the zymotic fluid of remembering effect;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; powder sieves and obtains dried particles by 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 with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes; be pressed into tablet, every 0.35g.
Embodiment 2
1, asparagus solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 80%, soybean 16.5%, walnut kernel 1%, fructus alpiniae oxyphyllae 1%, ginkgo leaf 1%, land plaster 0.5%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 60%; 121 ℃ of sterilizing 1h, obtain property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of asparagus ACCC50424 (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 asparagus ACCC50424 liquid spawn is property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 30 days has improvement memory effect for 20 ℃;
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 ℃ of sterilizing 30min, obtain fermentation medium;
2. by the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style, be: CGMCCNO.5655) liquid spawn (28 ℃ of standing cultivation 48h of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that the standing cultivation of 1%, 28 ℃ of fermentation medium volume 72h preparation has releasing the pressure and improves the zymotic fluid of remembering effect;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; powder sieves and obtains dried particles by 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 with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes; with capsule filling machine encapsulating capsule, every 0.35g.
Embodiment 3
1, asparagus solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 75%, soybean 15%, walnut kernel 3%, fructus alpiniae oxyphyllae 3%, ginkgo leaf 3%, land plaster 1%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 63%; 121 ℃ of sterilizing 1h, obtain property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of asparagus ACCC50424 (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 asparagus ACCC50424 liquid spawn is property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 20 days has improvement memory effect for 25 ℃;
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 ℃ of sterilizing 30min, obtain fermentation medium;
2. by the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style, be: CGMCCNO.5655) liquid spawn (28 ℃ of standing cultivation 48h of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that the standing cultivation of 1%, 32 ℃ of fermentation medium volume 48h preparation has releasing the pressure and improves the zymotic fluid of remembering effect;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; powder sieves and obtains dried particles by 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 with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes; be pressed into tablet, every 0.35g.
Embodiment 4
1, asparagus solid fermentation:
1. the making of property of medicine culture matrix
Formula: wholegrain barley 75%, soybean 15%, walnut kernel 3%, fructus alpiniae oxyphyllae 3%, ginkgo leaf 3%, land plaster 1%;
Compound method: press formula rate each component is mixed, pulverized, adding water to moisture weight content is 62%, 121 ℃ of sterilizing 1h, obtains property of medicine culture matrix;
2. inoculation fermentation
The liquid spawn of asparagus ACCC50424 (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 asparagus ACCC50424 liquid spawn is property of medicine culture matrix), cultivate the medicinal fungal substance that preparation in 20 days has improvement memory effect for 25 ℃;
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 ℃ of sterilizing 30min, obtain fermentation medium;
2. by the Lactobacillus plantarum LW106(deposit number screening from Pickles, Sichuan Style, be: CGMCCNO.5655) liquid spawn (28 ℃ of standing cultivation 48h of MRS culture medium) is inoculated into 1. middle fermentation medium in 1% ratio, the volume adding that is Lactobacillus plantarum is that the standing cultivation of 1%, 32 ℃ of fermentation medium volume 48h preparation has releasing the pressure and improves the zymotic fluid of remembering effect;
3, first the zymotic fluid that prepared by the 2nd step filters, concentrates, is spray dried to powder; powder sieves and obtains dried particles by 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 with respect to 0.1% magnesium stearate lubricant of whole grain gained particle weight and in multi-dimensional movement mixer, fully mixes 5 minutes; with capsule filling machine encapsulating capsule, every 0.35g.
By toxicity and function test, further illustrate the present invention below.Due to capsule of the present invention and tablet place formulation, auxiliary material different outside, both primary raw materials, every day, dose, functional component were all identical, did not affect safety and the function of product.Therefore following, for capsule, carry out toxicity and function test.
Test example 1
Toxicological test:
Test objective: whether check 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 of 0.35g (body weight is by 60kg), amounts to dosage 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 ℃, 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, with sterilized water, be mixed with 0.500g/mL, dosage for 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 that embodiment 4 makes and be mixed with 0.5 with sterilized water, 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 capsule 's content that embodiment 4 makes with sterilized water be 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: given the test agent is all greater than 20.0g/kgbw to the maximal tolerance dose of female male chmice acute per os (MTD), 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
The function test of releasing the pressure:
Test objective: whether check product has the function of releasing the pressure to human body
The capsule that test material: embodiment 4 makes.
Sample: the oral recommended amounts of human body is every day 2 times, each 2, capsule No.1, No. 2, the packing of the two, outward appearance are basic identical, but one of them capsule making for embodiment 4, another is placebo.
Experimenter selects: experimenter is the scientific research personnel that operating pressure is large, and the age is in 35-55 year, and male or female allly meets above-mentioned condition and voluntary participation guarantee that cooperation person all can include test in.Get rid of following personnel: to health food allergy sufferers; Take in a short time the article relevant to tested function, have influence on the judgement to result.
Test method: when human body is in the state of pressure, will secrete cortisol in saliva, therefore can evaluate the effect that tested material is released the pressure as pressure index by take saliva cortisol.Experimenter is divided into test-meal group and placebo group at random, and take every day by RD, takes continuously 4 weeks.Duration of test does not change original eating habit, normal diet.As shown in Figure 2, Fig. 2 is the rate of change (starting point 100%) of experimenter's saliva cortisol to result.As shown in Figure 2, compare with control group, test-meal group experimenter saliva cortisol increases and significantly reduces (p<0.05), illustrates that the capsule that embodiment 4 makes has the effect of releasing the pressure.
Test example 3
Improve memory function test:
Test objective: whether check product has the function of improving memory to human body
The capsule that test material: embodiment 4 makes.
Sample: the oral recommended amounts of human body is every day 2 times, each 2, capsule No.1, No. 2, the packing of the two, outward appearance are basic identical, but one of them capsule making for embodiment 4, another is placebo.
Experimenter selects: failure of memory person, and age 25-50 year, select the volunteer who meets experimental condition to take tested material 45 days, 106 of experimenters, are divided into each 53 of test-meal group, control groups at random, and when test-meal finishes, test-meal group 1 people and control group 2 people do not participate in inspection.Before and after test-meal, experimenter's spirit diet, stool and urine situation no abnormality seen.
Test method: according to the check of < < health food and assessment technique standard > > (2003 editions) evaluation criterion, selecting memory quotient is the effect that index is evaluated the improvement memory of tested material.Adopt counter point between Self-control method and group, duration of test does not change original eating habit, normal diet.Result is as shown in table 1.
The impact of table 1 product on memory quotient
? Before test After test
Control group 87.13±19.82 91.16±20.93
Test group 87.09±21.02 99.33±18.68
As shown in Table 1, memory quotient and the control group comparison of test group before test-meal, no significant difference (P>0.05), the memory quotient of test group and control group relatively increase significantly (P<0.05) after test-meal, and during test-meal, do not find obvious adverse reaction, the effect that capsule that example 4 makes has auxiliary improvement of memory is executed in result judgement thus.

Claims (10)

1. release the pressure and improve the two-step fermentation preparation method of the functional food of memory, it is characterized in that, comprising the steps:
(1) asparagus solid fermentation: take containing the natural constituents that is improved memory effect is property of medicine culture matrix, inoculation flammulina velutipes liquid strains carries out two-way solid fermentation preparation and has the medicinal fungal substance that improves memory effect;
(2) lactic acid bacteria liquid submerged fermentation: the medicinal fungal substance of step (1) of take is 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 of releasing the pressure, and preparation has releasing the pressure simultaneously and improves the zymotic fluid of memory effect;
(3) adopt the zymotic fluid of step (2), be prepared into releasing the pressure and improving the functional food of memory of different dosage form.
2. the two-step fermentation preparation method who releases the pressure and improve the functional food of memory according to claim 1, is characterized in that, in step (1), the inoculum concentration of described flammulina velutipes liquid strains is 2%-10%.
3. the two-step fermentation preparation method who releases the pressure and improve the functional food of memory according to claim 1, 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%, soybean 10%-20%, walnut kernel 1%-3%, fructus alpiniae oxyphyllae 1%-3%, ginkgo leaf 1%-3%, land plaster 0.5%-1.5%, each component is mixed, pulverized, then adding water to moisture weight content is 60%-65%, 116 ℃ of-126 ℃ of sterilizing 0.5h-1.5h.
4. the two-step fermentation preparation method who releases the pressure and improve the functional food of memory according to claim 3, 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%, soybean 15%, walnut kernel 3%, fructus alpiniae oxyphyllae 3%, ginkgo leaf 3%, land plaster 1%, each component is mixed, pulverized, then adding water to moisture weight content is 60%-65%, 121 ℃ of sterilizing 1h.
5. the two-step fermentation preparation method who releases the pressure and improve the functional food of memory according to claim 1, is characterized in that, in step (1), the condition of described two-way solid fermentation is: fermentation temperature is at 20-25 ℃, and fermentation time is at 20-30 days.
6. the two-step fermentation preparation method who releases the pressure and improve the functional food of memory according to claim 1, 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, medicinal fungal substance, dusty yeast and sodium glutamate is water-soluble, add acetic acid and sodium acetate and be mixed with the buffer solution that pH is 5-6,116 ℃ of-126 ℃ of sterilizing 20min-40min.
7. the two-step fermentation preparation method who releases the pressure and improve the functional food of memory according to claim 1, 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 who releases the pressure and improve the functional food of memory according to claim 1, is characterized in that, in step (2), the condition of described liquid deep layer fermenting is: fermentation temperature is at 28-32 ℃, and fermentation time is at 48-72h.
9. the two-step fermentation preparation method who releases the pressure and improve the functional food of memory according to claim 1, is characterized in that, in step (2), the described formulation of releasing the pressure and improving the functional food of memory is tablet or capsule.
10. that according to the preparation method described in claim 1~9 any one, prepares releases the pressure and improves the functional food of memory.
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CN109223668A (en) * 2018-09-30 2019-01-18 上海清轩生物科技有限公司 A kind of preparation method and application of the two-step fermentation product with skin repair
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CN113229494A (en) * 2021-04-14 2021-08-10 四川省农业科学院土壤肥料研究所 Needle mushroom water extract with high gamma-aminobutyric acid content and preparation method thereof

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