CN108185140A - Pueraria lobata composite fermentation object and its preparation method and application - Google Patents
Pueraria lobata composite fermentation object and its preparation method and application Download PDFInfo
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- CN108185140A CN108185140A CN201810086336.2A CN201810086336A CN108185140A CN 108185140 A CN108185140 A CN 108185140A CN 201810086336 A CN201810086336 A CN 201810086336A CN 108185140 A CN108185140 A CN 108185140A
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- pueraria lobata
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- 230000001376 precipitating effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 230000003014 reinforcing effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 239000000932 sedative agent Substances 0.000 description 1
- 230000001624 sedative effect Effects 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- 235000014214 soft drink Nutrition 0.000 description 1
- 239000007909 solid dosage form Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/076—Poria
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/481—Astragalus (milkvetch)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/488—Pueraria (kudzu)
-
- 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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- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
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- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/249—Thermophilus
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- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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Abstract
The invention discloses a kind of pueraria lobata composite fermentation objects and its preparation method and application.The fermentation of different condition three times is carried out using the culture medium of the object of complex enzyme hydrolysis containing pueraria lobata, gained fermentate progress separation of solid and liquid is then obtained into fermenation raw liquid, as pueraria lobata composite fermentation object.The preparation method fermentation time of the present invention is short, so as to be more conducive to industrialized production.In addition, the fermentation method of the present invention can also improve the active constituent in gained fermentate, drug effect, particularly Antidiarrheic effect are significantly improved.
Description
Technical field
The present invention relates to a kind of pueraria lobata composite fermentation objects and its preparation method and application, especially a kind of that there is anti diar rhea to make
Pueraria lobata composite fermentation object and its preparation method and application.
Background technology
Pueraria lobata isoflavone-containing ingredient Puerarin, Puerarin xyloside, daizeol, daidzin and cupreol, flower
Raw acid, and containing much starch.Exterior syndrome fever, stiff nape and back, measles without adequate eruption, pyreticosis is thirsty, the deficiency of Yin is quenched one's thirst, heat for treating for pueraria lobata
Purge heat dysentery, splenasthenic diarrhea.
Natural resources of Chinese medicinal materials is utilized in order to deeply develop, has been carried out the research of herb fermenting technology in recent years.For example,
CN104606487A discloses a kind of fermented tcm preparation, wherein Chinese medicine material be rhizoma alismatis, root of Chinese trichosanthes, the coptis, Radix Codonopsis, pueraria lobata,
Sealwort, Cortex Phellodendri, balsam pear, the fruit of Chinese wolfberry, Schisandra chinensis, ganoderma lucidum, Poria cocos, earthworm, Chinese yam, rhizoma atractylodis, stiff silkworm, umbellate pore furgus, Rhizoma Atractylodis Macrocephalae, is known Radix Astragali
It is female.The fermented tcm preparation provided in the document has good effect in terms for the treatment of, healing diabetes.
For another example, CN106306512 A disclose a kind of fermentation of Chinese herbal medicine feed addictive for treating diarrhea of pigs, and Chinese herbal medicine is
Pueraria lobata, wild chrysanthemum, honeysuckle, rhizoma anemarrhenae, the fruit of glossy privet, Radix Astragali, corydalis tuber, Cortex Magnoliae Officinalis, ginkgo leaf, garden burnet, grass-leaved sweetflag, prepared fleece flower root,
Radix Rehmanniae, Chinese mahonia, Chinese violet, hairyvein agrimony, Semen Cuscutae, radix scrophulariae, paniculate swallowwort, Poria cocos, radix bupleuri, cloves and rhizoma alismatis.The addition
Agent has the function of very strong animal disease resistant, promotes growth of animal, improve survival rate height, lower diarrhea rate.
Although above-mentioned document has carried out fermentation research for the compound containing pueraria lobata, in these technologies or raw material into
Divide complicated or anti diar rhea curative effect insufficient.
Therefore, pueraria lobata resource how is further developed and used, it is a subject for being worth research to improve pueraria lobata utilization rate.
Invention content
The purpose of the present invention is to provide the preparation methods of a kind of pueraria lobata composite fermentation object or pueraria lobata composite fermentation dry powder, should
Method can ferment using only three kinds of Chinese medicine materials, and fermentation time is short, and gained fermentate drug effect greatly promotes.This hair
Bright another object is to provide a kind of pueraria lobata composite fermentation product have improved anti-CaM antibody.Another mesh of the present invention
The purposes for being above-mentioned pueraria lobata composite fermentation product.The purpose of the present invention is what is be achieved through the following technical solutions.
The first aspect of the present invention provides a kind of preparation method of pueraria lobata composite fermentation object, includes the following steps：
First fermentation step：10~20h of anaerobic fermentation is carried out in the medium at 26~32 DEG C using saccharomycete, is obtained
First fermentate, wherein the culture medium includes pueraria lobata complex enzyme hydrolysis object；
Second fermentation step：The temperature of first fermentate is risen to 33~40 DEG C, and at such a temperature using lactic acid bacteria into
Row 30~40h of standing for fermentation, obtains the second fermentate；
Third fermentation step：The temperature of second fermentate is risen to 50~70 DEG C, and at such a temperature standing for fermentation 25~
35h obtains third fermentate；
Solid-liquid separation step：Gained third fermentate progress separation of solid and liquid is obtained into fermenation raw liquid, the as compound hair of pueraria lobata
Ferment object.
Preparation method according to the present invention, it is preferable that the pueraria lobata complex enzyme hydrolysis object is by using pectase, fiber
Plain enzyme, zytase and trypsase are digested at a temperature of 6.0~7.2 pH value and 45~60 DEG C containing pueraria lobata, Radix Astragali and Fu
2~6h of water slurry of Siberian cocklebur is made.
Preparation method according to the present invention, it is preferable that the dosage of the pectase is water slurry total weight
0.05~0.3wt%, the dosage of the cellulase are 0.05~0.3wt% of water slurry total weight, the use of zytase
It is the 0.05 of water slurry total weight to measure as 0.05~0.3wt% of water slurry total weight and the dosage of the trypsase
~0.15wt%.
Preparation method according to the present invention, it is preferable that pectase, cellulase, zytase and trypsase
Weight ratio is 1:1:1:1.
Preparation method according to the present invention, it is preferable that described containing in the water slurry of pueraria lobata, Radix Astragali and Poria cocos,
Pueraria lobata is 30~55 parts by weight, Radix Astragali is 30~40 parts by weight, Poria cocos is 10~20 parts by weight, and the content of pueraria lobata is more than Radix Astragali
Content.
Preparation method according to the present invention, it is preferable that the culture medium further includes 0.1~0.5wt% yeast
Cream, 1~5wt% ammonium salts, 0.01~0.2wt% magnesium salts, 0.3~0.8wt% phosphate and 0.01~0.2wt% zinc salt；With
Upper weight percent is all based on the total weight of the culture medium.
Preparation method according to the present invention, it is preferable that the inoculum concentration of saccharomycete described in the first fermentation step is every
Culture medium 1 × 10 described in ml7Cfu~9 × 107Cfu, the inoculum concentration of lactic acid bacteria described in the second fermentation step are training described in per ml
Support base 1 × 106Cfu~9 × 106cfu。
The second aspect of the present invention provides a kind of preparation method of pueraria lobata composite fermentation dry powder, including using the present invention
Preparation method described in first aspect obtains pueraria lobata composite fermentation object, is then concentrated and dried the pueraria lobata composite fermentation object
Fermentation dry powder is made.
The third aspect of the present invention provides a kind of pueraria lobata composite fermentation product, is according to first aspect present invention
The pueraria lobata composite fermentation that preparation method described in the pueraria lobata composite fermentation object or second aspect of the present invention that preparation method obtains obtains
Dry powder.
The fourth aspect of the present invention, the pueraria lobata composite fermentation product for providing the present invention are preparing the medicine with anti-CaM antibody
Purposes in object, food or feed.
The present invention combines the mode of forced fermentation three times by using enzymolysis so that fermentation time is reduced, so as to more added with
Conducive to industrialized production.In addition, the forced fermentation of the present invention can also improve the active constituent in gained fermentate, medicine is significantly improved
Effect, particularly Antidiarrheic effect.
Specific embodiment
With reference to specific embodiment, the present invention is further illustrated, but protection scope of the present invention is not limited to
This.
" pueraria lobata composite fermentation object " of the present invention refers to carry out biofermentation system using the herbal mixture comprising pueraria lobata
Standby obtained fermentate is sometimes referred to as used as " pueraria lobata composite enzyme " or referred to as " composite fermentation object " or " compound ferment herein
Element ".
The present invention is directed to the problem of drug effect is not high when utilizing kudzuvine root for treating diarrhea, it is proposed that based on enzymolysis Gegen Compound Chinese medicine
Ingredient, and combine fermentation three times and carry out the scheme of intensive treatment, and the present invention is completed based on the program.Specifically, it is of the invention
Including the following contents.
The first aspect of the present invention provides a kind of preparation method of pueraria lobata composite fermentation object, including fermentation step three times
And solid-liquid separation step.The following detailed description of each step.
<First fermentation step>
First fermentation step be saccharomycete anaerobic fermentation step, including the use of saccharomycete at 26~32 DEG C in the medium
10~20h of anaerobic fermentation is carried out, obtains the first fermentate, wherein the culture medium includes pueraria lobata complex enzyme hydrolysis object.
It ferments to generate to pueraria lobata complex enzyme hydrolysis object under anaerobic using saccharomycete and intestinal beneficial bacterium is proliferated
There is effective metabolite of facilitation.The inoculum concentration of saccharomycete is culture medium 1 × 10 described in per ml in first fermentation7Cfu~9
×107Cfu, preferably 1.5 × 107Cfu~7 × 107Cfu, more preferable 3 × 107Cfu~6 × 107cfu.Ability can be used in saccharomycete
Any strain known to domain, it is especially not specific.It is preferred that the high activity dried yeast produced by Angel Yeast Co., Ltd, the bacterium
Kind is particularly advantageous for the biological conversion of kudzu vine root.
The fermentation temperature of first fermentation is 26~32 DEG C, preferably 27~31 DEG C, more preferably 28 DEG C, this temperature range one
Aspect is conducive to the fermentation of saccharomycete, and also helps the biological conversion of kudzu vine root.If the first fermentation temperature
It is excessively high, then it can influence the activity of saccharomycete.
The fermentation time of first fermentation is 10~20h, more preferably preferably 11~18h, 16h.This fermentation time can be with
So that the effect of saccharomycete gives full play to, high active ingredient content is high in gained fermentate.
Culture medium in first fermentation step need to include pueraria lobata complex enzyme hydrolysis object.Pueraria lobata complex enzyme hydrolysis object is passes through enzymatic treatment
Make to be conducive to the ingredient release of fermentation or the enzymatic treatment object being unfavorable for after the ingredient breakdown of fermentation.Preferably, pueraria lobata complex enzyme hydrolysis
Object is to be mixed by using what the combined treatment Gegen Compound medicinal material of pectase, cellulase, zytase and trypsase obtained
Close object.Four kinds enzyme can efficiently effect, promotion digest simultaneously under the conditions of herein described, and unfavorable shadow is not present between enzymatic activity
It rings.
Gegen Compound medicinal material includes pueraria lobata and other Chinese medicines, other Chinese medicines are preferably Radix Astragali and/or Poria cocos.Gegen Compound medicine
Material is the combination of pueraria lobata, Radix Astragali and Poria cocos.According to embodiment of the present invention, Gegen Compound medicinal material is only by pueraria lobata, Radix Astragali
It is formed with Poria cocos.
In the present invention, pueraria lobata (Pueraria lobata (Willd.) ohwi) isoflavone-containing ingredient Puerarin, Puerarin
Xyloside, daizeol, daidzin and cupreol, arachidic acid, and containing much starch.Table is used it in Traditional Chinese Medicine
Demonstrate,prove fever, stiff nape and back, measles without adequate eruption, pyreticosis is thirsty, the deficiency of Yin is quenched one's thirst, heat is purged heat dysentery, splenasthenic diarrhea.The present invention passes through with pueraria lobata
For monarch drug in a prescription, reasonable compatibility enhances its splenasthenic diarrhea.
In the present invention, Radix Astragali (Astragalus membranaceus (Fisch.) Bunge.) is containing saponin, sucrose, more
The various trace elements such as sugared, a variety of amino acid, folic acid and selenium, zinc, copper.Traditional Chinese Medicine think the effect of Radix Astragali have invigorating qi for consolidating superficies,
The effect of inducing diuresis for removing edema, detoxification, myogenic, suitable for spontaneous perspiration, night sweat, blood-arthralgia, edema, un-festering of ulcer or bursts and does not hold back etc. diseases long.This
Radix Astragali can be selected arbitrarily in invention, be not particularly limited.It is preferred that preparing astragalus membranaceus because its with pueraria lobata more can gain it is complementary, improve medicine
Effect.
In the present invention, Poria cocos (Poria cocos (Schw.) Wolf) containing pachymaran, B- pachyman, pachymic acid,
Lecithin and sterol etc..Traditional Chinese Medicine thinks that it has effects that internal heat anticance, sharp aquation drink, strengthening the spleen and reducing phlegm, antitoxic heart-soothing and sedative.
Using Radix Astragali as minister in three kinds of medicinal materials, monarch's pueraria lobata is helped, then be aided with Poria cocos Ying Chen, monarch reasonable compatibility, Xiang Yuxuan
It takes the photograph, imperial spleen deficiency, stopping leak rush down.Preferably, pueraria lobata be 30~55 parts by weight, preferably 35~50 parts by weight；Radix Astragali is 30~40 weight
Part, preferably 32~48 parts by weight；Poria cocos be 10~20 parts by weight, preferably 12~18 parts by weight.Preferred pueraria lobata in the present invention
Content is more than the content of Radix Astragali.In fermentation, the combination of three kinds of medicinal materials can act synergistically, and Antidiarrheic effect is more excellent.
The condition of enzymatic treatment (being also referred to as sometimes herein " enzymolysis ") of the present invention is not particularly limited, for example, can be
2~6h of water slurry containing the compound medicine is digested at a temperature of 6.0~7.2 pH value and 45~60 DEG C.Preferably, enzymatic treatment
PH value for 6.5~6.9, more preferable 6.8, this pH value range can ensure that the various enzymes of the present invention keep high activity simultaneously.It is preferred that
Ground, the temperature of enzymatic treatment is 50~60 DEG C, more preferable 50 DEG C.Enzyme in the present invention need to be resistant to the enzyme of higher temperature, because too low
Hydrolysis temperature be unfavorable for the release of anti diar rhea ingredient.Enzyme processing time is preferably 3~5 hours, such as 4.5 hours.
Pectase during enzymatic treatment, cellulase, zytase and trypsase dosage, the total weight based on suspension,
It may respectively be 0.05~0.3wt%.It is preferred that be respectively 0.06~0.2wt%, respectively more preferably 0.08~0.15wt%, it is also excellent
Choosing is respectively 0.1~0.12wt%, and the dosage of trypsase can be 0.03~0.2wt%, preferably 0.05~0.15wt%,
More preferably 0.1~0.15wt%.Four kinds of enzymes are combined as that fermented and cultured provides amino acid, monosaccharide, oligosaccharides and other are beneficial
The raw materials such as ingredient.The weight ratio of enzyme is not especially specific, for example, pectase, cellulase, zytase and trypsase weight
Than that can be 1:1:1:1.
Herb or root are may respectively be for each medicinal material in the Gegen Compound medicinal material of enzymatic treatment, this is not particularly limited.
Preferably, it is powder for the pueraria lobata of enzymatic treatment, Radix Astragali and Poria cocos, the fineness of powder is not particularly limited, preferably thinner powder
End, so as to be more advantageous to the progress of enzymolysis.Preferably, above-mentioned three kinds of powder and water are mixed with water slurry, used it for
Enzymatic treatment.The total weight of wherein three kinds powder and the weight ratio of water are not particularly limited, and preferably 1:5~15, for example, 1:10.Water can
Use pure water, deionized water and distilled water.The pH of water is preferably 6.0~6.5, the water in the range of this pH be more advantageous to enzymolysis into
Row.
Culture medium in first fermentation step also may include nutriment.Preferably, nutriment may be selected from yeast extract, ammonium
At least one of salt, zinc salt, magnesium salts and phosphate ingredient.Total weight based on culture medium, culture medium may include 0.1~
0.5wt% yeast extracts, 1~5wt% ammonium salts, 0.01~0.2wt% magnesium salts, 0.3~0.8wt% phosphate and 0.01~
The zinc salt of 0.2wt%.Commercially available yeast extract may be used.The dosage of yeast extract is preferably 0.2~0.3wt%.Ammonium salt can select
From at least one of ammonium sulfate, ammonium nitrate or ammonium carbonate, preferably ammonium sulfate.The dosage of ammonium salt is preferably 1.5~
3.5wt%, more preferably 1.5~2.5wt%.Magnesium salts can be in magnesium sulfate, magnesium chloride, magnesium nitrate, magnesium carbonate at least
One kind, preferably magnesium sulfate.The dosage of magnesium salts is preferably 0.03~0.15wt%, more preferably 0.05~0.1wt%.Phosphate
Sodium phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, calcium phosphate, phosphoric acid can be selected from
At least one of magnesium, preferably potassium dihydrogen phosphate.Phosphatic dosage is preferably 0.35~0.65wt%, and more preferably 0.35
~0.5wt%.Zinc salt can be selected from least one of zinc sulfate, zinc chloride, zinc nitrate, and preferably zinc chloride.The battalion
Foster substance can include the zinc salt of 0.03~0.2wt%, preferably 0.05~0.15wt%, more preferably 0.08~0.1wt%.
Nutriment and ratio in above range, be more advantageous to meeting saccharomycete, lactic acid bacteria fermented and cultured nutrient demand,
And fermentation time can be reduced.
The nutriment of the present invention can also contain sylvite.The sylvite and phosphate are preferably same substance, such as phosphoric acid
Potassium, dipotassium hydrogen phosphate or potassium dihydrogen phosphate, preferably potassium dihydrogen phosphate.
<Second fermentation step>
The step of second fermentation step carries out forced fermentation for addition lactic acid bacteria, including the temperature of the first fermentate is risen to
33~40 DEG C, and 30~40h of standing for fermentation is carried out using lactic acid bacteria at such a temperature, obtain the second fermentate.Second fermentate
For forced fermentation object or strengthen ferment.It not only has the general utility functions of common ferment, but also effect is especially for diarrhea
Effect is stronger.
The lactic acid bacteria of the present invention can be selected from the one or more of lactobacillus bulgaricus and streptococcus thermophilus, preferably protect
Add the mixture of Leah lactobacillus and streptococcus thermophilus.The weight ratio of lactobacillus bulgaricus and streptococcus thermophilus can be 1:10
~10:1, preferably 1:5~5:1, more preferably 1:1.The inoculum concentration of lactic acid bacteria is culture medium 1 × 10 described in per ml6Cfu~9
×106Cfu, preferably 2 × 106Cfu~8 × 106Cfu, more preferable 5 × 106Cfu~8 × 106cfu.The inoculum concentration of lactic acid bacteria needs big
The inoculum concentration of saccharomycete when first ferments.If the inoculum concentration of lactic acid bacteria is too low, first fermentation when saccharomycete amount it is opposite
Excessively, the proliferation of lactic acid bacteria is influenced, and then influences the reinforcing effect of fermentate.
The temperature of second fermentation step need to be higher than the temperature of the first fermentation step, more excellent preferably in the range of 33~40 DEG C
Select 35~39 DEG C, even more preferably 37 DEG C.Such temperature simulates work of the lactic acid bacteria in human body close to human body environment's temperature
With temperature, so as to which the active constituent for being converted into or being discharged from Chinese medicine is more advantageous to absorption of human body.
The time of second fermentation is not particularly limited, and is under normal conditions 30~40h, preferably 35~39h, more preferably
32h.Fermentation time is long, and the rate of bioconversion can become to decline, while can generate other by-products, these by-products are unfavorable
In the performance of drug effect.On the other hand, if fermentation time is too short, react insufficient, some drugs active ingredient not yet generates,
So as to which the drug effect for making gained fermentate reduces, and can cause the wasting of resources.
Second fermentation needs to be left to ferment.Standing for fermentation is more closely similar to the environment of internal beneficial flora, thus is preferred.
<Third fermentation step>
The step of third fermentation step is saccharomycete and lactic acid bacteria double bacterium hot fermentations, including by the temperature of the second fermentate
50~70 DEG C are risen to, and is left to ferment 25~35h at such a temperature, obtains third fermentate.
By the way that saccharomycete and lactic acid bacteria is made to be carried out at the same time and continue to ferment at 50~70 DEG C, can more effectively promote effectively into
The content divided quickly generates in a short time.Preferably 55~65 DEG C, more preferable 65 DEG C of the range of second fermentation temperature.In the temperature
Under, 20~35h is only left to ferment, preferably 21~32h, more preferable 26~30h can obtain having the third of required performance to ferment
Object substantially reduces the fermentation time in conventional method.
<Solid-liquid separation step>
Produced object solid residue when separation of solid and liquid is for removing fermentation, and recycle fermenation raw liquid.Separation of solid and liquid can be used
Any known method, including but not limited to centrifuge, filter, precipitating etc..Preferably, centrifugation mode can be used in separation of solid and liquid,
For example, it is detached using centrifuge.The actual conditions of centrifugation are not particularly limited, and can be adjusted as needed and suitably.Pass through
Isolated fermenation raw liquid, as pueraria lobata composite fermentation object.
The second aspect of the present invention provides a kind of preparation method of pueraria lobata composite fermentation dry powder, using above-mentioned preparation method
Pueraria lobata composite fermentation object is obtained, then carries out being concentrated and dried obtained fermentation dry powder by the pueraria lobata composite fermentation object.It is dry and/or
Concentration step can be used for removing influence of the medium molecular chaperones for drug effect.Preferably, include before the drying step
Concentration step.Yeast powder is can obtain by concentrating and drying.The method that decompression may be used is concentrated and is dried, here no longer
It repeats.
The third aspect of the present invention provides a kind of pueraria lobata composite fermentation product, to pass through preparation side of the present invention
The pueraria lobata composite fermentation object or pueraria lobata composite fermentation dry powder that method is prepared.According to embodiment of the present invention, the hair
The water content of ferment dry powder is less than 5wt%, and the vigor of preferably shorter than 4.5wt%, superoxide dismutase SOD is more than or equal to 420 ±
5U/g, viable count of lactobacillus are more than or equal to 3.3 ± 0.5 × 108CFU/g, puerarin content are more than 5wt%.
The fourth aspect of the present invention provides pueraria lobata composite fermentation product and is preparing drug, food with anti-CaM antibody
Or the purposes in feed addictive.Wherein additive amount of the pueraria lobata composite fermentation object in drug, food or feed addictive be not special
It does not limit.
Example as drug includes but not limited to pulvis, pill solid dosage forms, further includes suspending agent, intermixture, emulsification
The liquid dosage forms such as agent.Example as food includes but not limited to candy, beverage (such as all kinds of soft drinks), bread, biscuit, wine
And all kinds of health products etc..Example as feed includes but not limited to feed addictive.Feed addictive refers to give birth in feed
The a small amount of or micro substance added during production processing, use, dosage is seldom in feed but effect is notable.
The preparation method of embodiment 1- pueraria lobata composite fermentation objects
1) it crushes：Pueraria lobata, Radix Astragali and Poria cocos be crushed into No. 2 sieves of pharmacopeia respectively, by 50 parts by weight of pueraria lobata, 35 weight of Radix Astragali
Part and the ratios of 15 parts by weight of Poria cocos raw material is mixed, then add in 10 times of material qualities than pure water, suspension is made.
2) preparation of enzymolysis liquid：Total weight based on suspension adds pectase (500,000 U/ of unit of activity of 0.1wt%
G), the cellulase (500,000 U/g of unit of activity) of 0.1wt%, the zytase (500,000 U/g of unit of activity) of 0.1wt% and pancreas
The dosage of protease is digested for 0.1wt% (500,000 U/g of unit of activity).PH is 6.8, and temperature is 52 DEG C, and enzymolysis time is
4.5h。
3) nutriment is added in right amount：Yeast extract 0.3wt%, ammonium sulfate 1.5wt%, zinc chloride 0.05wt%, magnesium sulfate
0.05wt%, potassium dihydrogen phosphate 0.4wt%, fermentation substrate sterilizing.
4) it is inoculated with and ferments：Inoculation yeast bacterium 3 × 107Cfu/mL, under the conditions of 28 DEG C, anaerobic fermentation 16h, inoculating lactic acid bacterium
Leavening 5 × 106Cfu/mL, then under the conditions of 37 DEG C, is left to ferment 32h, and it is 65 DEG C finally to maintain temperature, stands 30h, into
Row fermentation.
5) it centrifuges：Centrifugal treating, isolated fermentate stoste and solid residue are carried out after fermentation.Fermentate stoste
As pueraria lobata composite fermentation object.
Embodiment 2- pueraria lobata composite fermentation powder
1) it crushes：Pueraria lobata, Radix Astragali and Poria cocos be crushed into No. 2 sieves of pharmacopeia respectively, by 50 parts by weight of pueraria lobata, 35 weight of Radix Astragali
Part and the ratios of 15 parts by weight of Poria cocos raw material is mixed, then add in 10 times of material qualities than pure water, suspension is made.
2) preparation of enzymolysis liquid：Add the pectase (500,000 U/g of unit of activity) of 0.1wt%, the cellulase of 0.1wt%
The zytase (500,000 U/g of unit of activity) of (500,000 U/g of unit of activity), 0.1wt% and the trypsase (vigor of 0.15wt%
500,000 U/g of unit) it is digested.PH is 6.8, and temperature is 50 DEG C, enzymolysis time 4.5h.
3) nutriment is added in right amount：Yeast extract 0.3wt%, ammonium sulfate 1.5wt%, zinc chloride 0.05wt%, magnesium sulfate
0.05wt%, potassium dihydrogen phosphate 0.4wt%, fermentation substrate sterilizing.
4) it is inoculated with and ferments：Inoculation yeast bacterium 6 × 107Cfu/mL, under the conditions of 28 DEG C, anaerobic fermentation 16h, inoculating lactic acid bacterium
Leavening 8 × 106Cfu/mL, then under the conditions of 37 DEG C, is left to ferment 32h, and it is 65 DEG C finally to maintain temperature, stands 30h, into
Row fermentation.
5) it centrifuges：Centrifugal treating, isolated fermentate stoste and solid residue are carried out after fermentation；
6) it concentrates：Fermentate stoste after separation is concentrated under reduced pressure, by being dried under reduced pressure to obtain yeast powder, wherein water contains
It measures as 4wt%.
Embodiment 3- pueraria lobata composite fermentation object beverages
By pueraria lobata composite fermentation object made from embodiment 1 and water with 1:6 weight ratio mixing, addition 15wt% sucrose,
0.5%wt citric acids, and-filling-cooling that carries out homogeneous-degassing-pasteurize and etc., it can obtain finished beverage.
Embodiment 4- pueraria lobata composite fermentation object wine
By pueraria lobata composite fermentation object made from embodiment 1 and 45 degree of high quality liquors, drinking water with 1:8:4 weight ratio is mixed
- filling-cooling is closed, and carry out homogeneous-degassing-pasteurize, can obtain pueraria lobata composite fermentation object blended liquor.
The variation of puerarin content before and after embodiment 5- fermentations
The preparation of reference substance solution：Precision weighs that Puerarin reference substance is appropriate, adds 30% ethyl alcohol that every 1ml is made containing 80 μ g
Solution to get.
The preparation of test solution：
Fermentate sample：The yeast powder 0.35g for weighing embodiment 2 (is equivalent to pueraria lobata 2.0g, Radix Astragali 1.4g and Poria cocos
0.6g), it is accurately weighed, it puts in conical flask with cover, precision adds in 30% ethyl alcohol 50ml, and weighed weight is heated to reflux 30 minutes, puts
It is cold, then weighed weight, the weight of less loss is supplied with 30% ethyl alcohol, is shaken up, filters, takes subsequent filtrate 1mL in 10mL volumetric flasks, is used
30% ethyl alcohol be settled to graduation mark to get.
Aqueous extracts sample：Pueraria lobata 2.0g, Radix Astragali 1.4g and Poria cocos 0.6g are taken, water impregnates 30min, decocts extraction 2 times, every time
10 times of amounts of amount of water, each 2 hours extraction times, filtering, filtrate merge, are concentrated into dry powder, put in conical flask with cover, and precision adds
Enter 30% ethyl alcohol 50ml, weighed weight is heated to reflux 30 minutes, lets cool, then weighed weight, and the weight of less loss is supplied with 30% ethyl alcohol
Amount, shake up, filter, take subsequent filtrate 1mL in 10mL volumetric flasks, with 30% ethyl alcohol be settled to graduation mark to get.
Assay method：
It is accurate respectively to draw reference substance solution and each 10 μ l of test solution, inject liquid chromatograph, measure to get.Color
Spectral condition：Mobile phase is first alcohol and water (25:75), Detection wavelength 250nm dissolves reference substance Puerarin with 30% ethanol solution.
Chromatographic column：Diamonsil C18 columns (250mm × 4.6mm, 5 μm), flow velocity 1.0mL/min, run time 30min, sample size 20
μL。
The measurement result of 1 fermentate sample of table and Aqueous extracts sample puerarin content
| Sample | For test liquid puerarin content mg/mL |
| Fermentate sample | 0.0452 |
| Aqueous extracts sample | 0.0286 |
Embodiment 6- anti-CaM antibody effect experiments
The preparation of drug：
Yeast powder：Example 2 is made yeast powder 3.5g and (is equivalent to pueraria lobata 20g crude drug in whole, Radix Astragali 14g crude drug in whole and Fu
Siberian cocklebur 6g crude drug in whole), it adds in distilled water and is settled to 120ml to get (calculating volume according to intragastric administration on mice volume 20ml/kg).
It is prepared by Aqueous extracts：Pueraria lobata 20g, Radix Astragali 14g and Poria cocos 6g are taken, water impregnates 30min, decocts extraction 2 times, adds water every time
10 times of amounts of amount, each 2 hours extraction times, filtering, filtrate merge, and rotation rotary evaporation concentration is settled to 120ml with distilled water,
To obtain the final product.
Root of kudzu vine extract：Pueraria lobata 40g is taken, water impregnates 30min, decocts extraction 2 times, and each 10 times of amounts of amount of water carry every time
Take time 2 h, filter, filtrate merge, rotation rotary evaporation concentration, with distilled water be settled to 120ml to get.
Radix Astragali extractive solution：Radix Astragali 40g is taken, water impregnates 30min, decocts extraction 2 times, and each 10 times of amounts of amount of water carry every time
Take time 2 h, filter, filtrate merge, rotation rotary evaporation concentration, with distilled water be settled to 120ml to get.
Poria cocos extracting solution：Poria cocos 40g is taken, water impregnates 30min, decocts extraction 2 times, and each 10 times of amounts of amount of water carry every time
Take time 2 h, filter, filtrate merge, rotation rotary evaporation concentration, with distilled water be settled to 120ml to get.
Folium sennae suspension：Before use by dry blade (removing vein), add in boiling distilled water and grind and added after uniform cold
Distilled water is configured to 8% folium sennae suspension.
Experimental method：
Mouse 60, half male and half female are taken, weight (20 ± 2.0) g, every group 10, distinguishes blank control group by random point 6 groups,
Yeast powder, Aqueous extracts group, pueraria lobata group, Radix Astragali group and Poria cocos group, successive administration 5d, respectively gavage give corresponding liquid, blank pair
It gives the physiological saline of same volume according to group, before last dose after fasting 12h, each group after 1h is administered distinguishes 8% folium sennae of gavage and mix
Suspension 0.3ml/ only, divides cage, and per one cage of mouse, blotting paper (filter paper) is laid under cage, records each animal defecation frequency, observation is following
Indices, and statistical analysis.
Loose stool rate：Every animal passage of loose stools number and total just the ratio between number, using every on filter paper or every heap excrement is primary as defecation.
Loose stools grade：It is defined the level with loose stools pollution filter paper area diameter (cm), is divided into level Four：1 grade (<10), 2 grades (1~
1.9), 3 grades (2~3), 4 grades (>3) series per a pile loose stools, is first counted during statistics one by one, then by all loose stools grades of the mouse
Number addition divided by loose stools number obtain the average series of loose stools
Scours index：The product of loose stool rate and loose stools average rank.
Table 2, anti diar rhea experimental result
| Group | Animal number of elements (n) | Dosage (g/kg) | Scours index |
| Blank control group | 10 | -- | 1.00±0.17△△ |
| Yeast powder group | 10 | 6.7 | 0.44±0.15** |
| Aqueous extracts group | 10 | 6.7 | 0.60±0.10**△ |
| Pueraria lobata group | 10 | 6.7 | 0.75±0.12**△△ |
| Radix Astragali group | 10 | 6.7 | 0.98±0.15△△ |
| Poria cocos group | 10 | 6.7 | 0.71±0.15**△△ |
Note：Compared with blank group, * P ＜ 0.05**P ＜ 0.01；Compared with yeast powder group, △ P ＜ 0.05, △ △ P ＜
0.01。
By experimental data statistical result it is found that compared with blank group, yeast powder group, Aqueous extracts group, pueraria lobata group, Poria cocos group
The conspicuousness of diarrhea of mouse index reduces (P ＜ 0.01).Compared with yeast powder, Aqueous extracts group, pueraria lobata group, Poria cocos group and Radix Astragali group
Spleen Scours index conspicuousness reduces (P ＜ 0.05).This show yeast powder group reduce the effect of diarrhea of mouse index better than Aqueous extracts group,
Pueraria lobata group, Poria cocos group and Radix Astragali group.
Present invention is not limited to the embodiments described above, in the case of without departing substantially from the substantive content of the present invention, this field skill
Any deformation, improvement, the replacement that art personnel are contemplated that each fall within the scope of the present invention.
Claims (10)
1. a kind of preparation method of pueraria lobata composite fermentation object, which is characterized in that include the following steps：
First fermentation step：10~20h of anaerobic fermentation is carried out in the medium at 26~32 DEG C using saccharomycete, obtains first
Fermentate, wherein the culture medium includes pueraria lobata complex enzyme hydrolysis object；
Second fermentation step：The temperature of first fermentate is risen to 33~40 DEG C, and quiet using lactic acid bacteria progress at such a temperature
30~40h of fermentation is put, obtains the second fermentate；
Third fermentation step：The temperature of second fermentate is risen to 50~70 DEG C, and be left to ferment 25~35h at such a temperature,
Obtain third fermentate；
Solid-liquid separation step：Gained third fermentate progress separation of solid and liquid is obtained into fermenation raw liquid, as pueraria lobata composite fermentation object.
2. preparation method according to claim 1, which is characterized in that the pueraria lobata complex enzyme hydrolysis object is by using pectin
Enzyme, cellulase, zytase and trypsase are digested at a temperature of 6.0~7.2 pH value and 45~60 DEG C containing pueraria lobata, Huang
2~6h of water slurry of stilbene and Poria cocos is made.
3. preparation method according to claim 2, which is characterized in that the dosage of the pectase is water slurry total weight
0.05~0.3wt%, the dosage of the cellulase is 0.05~0.3wt% of water slurry total weight, zytase
Dosage is 0.05~0.3wt% of water slurry total weight and the dosage of the trypsase is the 0.05 of water slurry total weight
~0.15wt%.
4. preparation method according to claim 3, which is characterized in that pectase, cellulase, zytase and tryptose
The weight ratio of enzyme is 1:1:1:1.
5. preparation method according to claim 1, which is characterized in that in the aqueous suspension containing pueraria lobata, Radix Astragali and Poria cocos
In liquid, pueraria lobata is 30~55 parts by weight, Radix Astragali is 30~40 parts by weight, Poria cocos is 10~20 parts by weight, and the content of pueraria lobata is big
In the content of Radix Astragali.
6. preparation method according to claim 1, which is characterized in that the culture medium further includes 0.1~0.5wt%
Yeast extract, 1~5wt% ammonium salts, 0.01~0.2wt% magnesium salts, 0.3~0.8wt% phosphate and 0.01~0.2wt% zinc
Salt；More than weight percent is all based on the total weight of the culture medium.
7. preparation method according to claim 1, which is characterized in that the inoculum concentration of saccharomycete described in the first fermentation step
For culture medium 1 × 10 described in every ml7Cfu~9 × 107Cfu, the inoculum concentration of lactic acid bacteria described in the second fermentation step is per ml institute
State culture medium 1 × 106Cfu~9 × 106cfu。
8. a kind of preparation method of pueraria lobata composite fermentation dry powder, which is characterized in that include the following steps：Using according to claim
1~7 any one of them preparation method obtains pueraria lobata composite fermentation object, and it is dry that the pueraria lobata composite fermentation object then is carried out concentration
Dry obtained fermentation dry powder.
9. a kind of pueraria lobata composite fermentation product, which is characterized in that it is according to claim 1~7 any one of them preparation side
The pueraria lobata composite fermentation dry powder that the pueraria lobata composite fermentation object or preparation method according to claim 8 that method obtains obtain.
10. pueraria lobata composite fermentation product according to claim 9 is preparing drug, food or feeding with anti-CaM antibody
Purposes in material.
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| CN115868630A (en) * | 2022-12-26 | 2023-03-31 | 山东金源中草药有限公司 | Plant functional beverage and preparation method thereof |
| CN116616366A (en) * | 2023-05-31 | 2023-08-22 | 广州大台农饲料有限公司 | Fermented nutritional composition for improving lactation capacity of sows as well as preparation method and application thereof |
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| CN104664270A (en) * | 2014-07-09 | 2015-06-03 | 普正药业股份有限公司 | Fresh and alive traditional Chinese medicine fruit-vegetable enzyme and preparation method thereof |
| CN105962360A (en) * | 2016-05-09 | 2016-09-28 | 山东祥维斯生物科技股份有限公司 | Method for producing microorganism ferment |
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| CN105962360A (en) * | 2016-05-09 | 2016-09-28 | 山东祥维斯生物科技股份有限公司 | Method for producing microorganism ferment |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111676168A (en) * | 2020-06-30 | 2020-09-18 | 大连澎立生物科技有限公司 | Nano-selenium detoxicant and preparation method thereof, detoxified biological protein selenium and preparation method and application thereof |
| CN111676168B (en) * | 2020-06-30 | 2022-01-25 | 大连澎立生物科技有限公司 | Nano-selenium detoxicant and preparation method thereof, detoxified biological protein selenium and preparation method and application thereof |
| CN112450345A (en) * | 2020-12-10 | 2021-03-09 | 重庆伍六奇农业科技开发有限公司 | Radix puerariae fermented beverage capable of improving intestinal function and preparation method thereof |
| CN115868630A (en) * | 2022-12-26 | 2023-03-31 | 山东金源中草药有限公司 | Plant functional beverage and preparation method thereof |
| CN116616366A (en) * | 2023-05-31 | 2023-08-22 | 广州大台农饲料有限公司 | Fermented nutritional composition for improving lactation capacity of sows as well as preparation method and application thereof |
| CN116616366B (en) * | 2023-05-31 | 2023-11-07 | 广州大台农饲料有限公司 | Fermented nutritional composition for improving lactation capacity of sows as well as preparation method and application thereof |
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