CN105559068A - Composition obtained by fermenting tetrastigma hemsleyanum diels et gilg. through edible and medicinal fungi and preparation method of composition - Google Patents
Composition obtained by fermenting tetrastigma hemsleyanum diels et gilg. through edible and medicinal fungi and preparation method of composition Download PDFInfo
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- CN105559068A CN105559068A CN201511027688.3A CN201511027688A CN105559068A CN 105559068 A CN105559068 A CN 105559068A CN 201511027688 A CN201511027688 A CN 201511027688A CN 105559068 A CN105559068 A CN 105559068A
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- radix tetrastigme
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Classifications
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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)
- A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K2236/10—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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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Zoology (AREA)
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Abstract
The invention discloses a composition obtained by fermenting tetrastigma hemsleyanum diels et gilg. through edible and medicinal fungi and a preparation method of the composition. The composition comprises a tetrastigma hemsleyanum diels et gilg. fermentation subcritical extract I and a streptomycete fermentation product III, and has a favorable function of reducing blood lipid. The preparation method comprises the following steps of inoculating Brazilian mushroom liquid seed fluid into tetrastigma hemsleyanum diels et gilg. solid state fermentation culture mediums for culturing, wherein the tetrastigma hemsleyanum diels et gilg. solid state fermentation culture mediums contain tetrastigma hemsleyanum diels et gilg. and bletilla striata, and then placing the cultured Brazilian mushroom liquid seed fluid in alternating magnetic field environment for culturing once again so as to obtain tetrastigma hemsleyanum diels et gilg. solid state fermentation products; then performing subcritical water extraction so as to obtain the tetrastigma hemsleyanum diels et gilg. fermentation subcritical extract I; inoculating streptomycete seed fluid into streptomycete liquid fermentation culture mediums for culturing so as to obtain the streptomycete fermentation products III, wherein the streptomycete liquid fermentation culture mediums contain extraction remnants II. According to the method disclosed by the invention, nutrient components and effective components in the tetrastigma hemsleyanum diels et gilg. can be effectively biodegraded and utilized, and the increase of the content and the activity of active components in finished products is facilitated.
Description
Technical field
The present invention relates to the processing and utilization field of radix tetrastigme, be specifically related to a kind of composition and method of making the same utilizing food medicine fungi fermentation radix tetrastigme to obtain.
Background technology
Radix tetrastigme (TetrastigmahemsleyanumDielsetGilg) is Vitaceae Tetrastigma plant, another name: gold thread hoist, stone mouse, hemsley rockvine root, genus Rhamnales, Vitaceae herbaceous species.Formal name used at school: tetratigma hemsleyanum is that the regional tradition such as Zhejiang, Fujian commonly uses green herbal medicine, best with the medicinal effects of underground block root and fruit.Among the people comparatively remote by radix tetrastigme treatment tumour in Zhejiang, radix tetrastigme is region medicinal materials, very rare, be distributed in the ground such as Zhejiang, Jiangxi, Fujian, Anhui, Guangdong, Guangxi, Yunnan, radix tetrastigme has heat radiation, regulates the flow of vital energy, invigorates blood circulation, tonifying spleen, detoxify, dispel the wind, the effect of anti-inflammatory, treatment infantile hyperpyrexia, pharyngo-laryngitis chronica, tonsillitis, hepatitis, pneumonia, gastritis, scrofula and the snake bite etc. of being used among the people has special effect.Modern pharmacological research shows that radix tetrastigme has the effects such as anti-inflammatory, analgesia and antipyretic, antitumor, antiviral, immunity moderation.
At present, the traditional extracting and developing method of the many employings of active component in radix tetrastigme obtains, for the Chinese medicine radix tetrastigme of prevention and therapy tumour disclosed in Chinese patent ZL200510050649.5, what it adopted is by radix tetrastigme is that the water of raw material and/or alcohol extract make active component.The method of the radix tetrastigme of supercritical carbon dioxide extracting disclosed in Chinese patent application CN201110264176.4 general flavone, by radix tetrastigme pulverizing medicinal materials 60-80 order, add supercritical carbon dioxide extracting tank, pass into liquid carbon dioxide and entrainer, under pressure 20-30MPa, temperature 40-50 DEG C condition, extraction 2-3 hour, in separator, extract is resolved in decompression, reclaim reagent, dry radix tetrastigme general flavone.Chinese patent ZL200810120750.7 discloses a kind of technique of high efficiency extraction flavone compound from radix tetrastigme, and its technological process is: raw material radix tetrastigme, air-dry, chopping, add water to extract in ultrasonic extractor, extract is concentrated into certain volume, then carry out adsorption bleaching with active carbon.Then cavitation suspension extraction separator is utilized, first be extracted with ethyl acetate, reclaim ethyl acetate, by aqueous solution part chloroform extraction, reclaim chloroform, by aqueous solution partial concentration extremely without organic reagent taste, by macroporous absorbent resin, use water and 50% ethanol elution successively, collect 50% ethanol eluate, recycling design, to dry, namely obtains Lemna paucicostata.
Containing polysaccharide, flavones, polyphenol isoreactivity composition and the nutritional labeling such as starch, protein in radix tetrastigme, in its active component technique of traditional extracting and developing, starch, protein, stimulating component Deng Cheng branch extract together, add the treatment step in separation, cause the active component content in final products on the low side, active on the low side.
Summary of the invention
The invention provides a kind of composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, in said composition the content of active component and activity high, there is good hypolipemic function.
Present invention also offers a kind of preparation method of the composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, utilize multiple fungi fermentation radix tetrastigme, contribute to the content and the activity that improve active component in final products, obtain the fermentation composition with hypolipemic function.
The present invention finds, according to radix tetrastigme nutritional labeling and active ingredient feature, by the specific fungi fermentation of the present invention and specific technique, and the bletilla striata is added in culture medium, effectively the nutritional labeling in radix tetrastigme and active ingredient can be carried out biodegradation and utilization, effectively the stimulating component in radix tetrastigme be removed and conversion, radix tetrastigme resource is fully utilized, contributes to the content and the activity that improve active component in final products.
The technical solution used in the present invention is:
A preparation method for the composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, comprises step:
(1) mushroom Agaricus blazei Murill got after activation is inoculated in liquid MRS culture medium and cultivates 2 days-8 days, collected by centrifugation first thalline; First thalline is suspended in the liquid MRS culture medium containing cholate and hatches 1h-20h in 18 DEG C-30 DEG C, collected by centrifugation second thalline; By the second thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the second thalline liquid, second thalline liquid is inoculated in the liquid MRS culture medium containing cholate and cultivates 1 day-10 days again in 18 DEG C-30 DEG C, obtain Brazilian mushroom liquid seeds liquid;
(2) streptomyces species got after activation is inoculated in liquid MRS culture medium and cultivates 3h-30h, collected by centrifugation the 3rd thalline, is suspended in by the 3rd thalline in the liquid MRS culture medium containing NaCl and hatches 0.5h-15h in 20 DEG C-30 DEG C, collected by centrifugation the 4th thalline; By the 4th thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the 4th thalline liquid, 4th thalline liquid is inoculated in the liquid MRS culture medium containing NaCl and cultivates 3h-30h in 20 DEG C-30 DEG C, obtain streptomycete seed liquor;
(3) by the Brazilian mushroom liquid seeds liquid of step (1) access radix tetrastigme solid-state fermentation culture medium, cultivate 2 days-20 days at 18 DEG C-30 DEG C, then be placed in alternating magnetic field environment and cultivate 5 days-30 days again, cultured products obtains radix tetrastigme product by solid-state fermentation after vacuum freeze drying, pulverizing; Containing radix tetrastigme and the bletilla striata in described radix tetrastigme solid-state fermentation culture medium;
(4) by the radix tetrastigme product by solid-state fermentation of step (3) through Subcritical Water Extraction, obtain radix tetrastigme fermentation subcritical abstraction thing I, after extraction, remaining residue is extracted residue II after vacuum freeze drying.
(5) by the streptomycete seed liquor of step (2) access streptomycete liquid fermentation medium, 10h-120h is cultivated at 20 DEG C-30 DEG C, centrifugal, obtain streptomycete fermentation product III; Containing extraction residue II in described streptomycete liquid fermentation medium;
The described composition utilizing food medicine fungi fermentation radix tetrastigme to obtain is radix tetrastigme fermentation subcritical abstraction thing I and streptomycete fermentation product III.
When the present invention carries out radix tetrastigme solid state fermentation, the direct Brazilian mushroom liquid seeds liquid adopted through hatching process containing liquid MRS culture medium two step of cholate, and the bletilla striata is added in culture medium, effectively the nutritional labeling in radix tetrastigme and active ingredient can be carried out biodegradation and utilization, effectively the stimulating component in radix tetrastigme be removed and conversion, contribute to the content and the activity that improve active component in final products.In step (4) and step (5), radix tetrastigme product by solid-state fermentation is carried out Subcritical Water Extraction, make that the activity of active ingredient in extract is at utmost retained, content is greatly improved; Again using the residue after extraction as the main culture medium of streptomycete liquid fermentation, and adopt the streptomycete seed liquor through hatching process containing liquid MRS culture medium two step of NaCl, recycled by integration, radix tetrastigme resource is fully utilized, further increases content and the activity of active component in final products.
In order to reach better effect, preferably:
In step (1), described is 0.05%-0.5% containing the mass percentage of cholate in the liquid MRS culture medium of cholate.
Described first thalline is 1:1-3 with the volume ratio of the liquid MRS culture medium containing cholate for first thalline that suspends, and the second thalline is 1:1-3 with the volume ratio for the liquid MRS culture medium of second thalline that suspends, and the inoculum concentration of the second thalline liquid is 5%.
In step (2), the mass percentage of the described liquid MRS NaCl in medium containing NaCl is 0.5%-8%.
Described 3rd thalline is 1:1-3 with the volume ratio of the liquid MRS culture medium containing NaCl for the 3rd thalline that suspends, and the 4th thalline is 1:1-3 with the volume ratio for the liquid MRS culture medium of the 4th thalline that suspends, and the inoculum concentration of the 4th thalline liquid is 3%.
In step (3), described radix tetrastigme adopts radix tetrastigme block root; The described bletilla striata adopts bletilla striata block root.Containing 6.5g-9.5g radix tetrastigme block root and 0.5g-3.5g bletilla striata block root in the every 1L of described radix tetrastigme solid-state fermentation culture medium.Described radix tetrastigme solid-state fermentation culture medium is made up of radix tetrastigme block root, bletilla striata block root and fermentation basal medium, and described fermentation basal medium adopts the fermentation basal medium of this area routine, can adopt commercially available prod, and existing compound method also can be adopted to prepare.
The preparation method of described radix tetrastigme solid-state fermentation culture medium, comprising: fresh radix tetrastigme block root, bletilla striata block root are rinsed well under running water, dry, cutting is 1cm-2cm lengthy motion picture shape or fritter; Again 6.5g-9.5g radix tetrastigme block root and 0.5g-3.5g bletilla striata block root fermentation basal medium are settled to 1L, mix, pH value nature, obtains radix tetrastigme solid-state fermentation culture medium.
The access amount of described Brazilian mushroom liquid seeds liquid is the 5%-25% of radix tetrastigme solid-state fermentation culture medium volume.
Current fungi solid-state fermentation technology ubiquity fermentation period is long, mycelial growth sprouts the problems such as slow, feature secondary metabolism metabolite content is low.The present invention finds alternating magnetic field treatment technology to be applied in solid state fermentation, by extraneous factor effective stimulus such as the suitable process of specific cultivation period, improve the growth metabolism of hypha,hyphae on radix tetrastigme culture base-material, shorten fermentation time, effectively promote the generation of secondary metabolite.In described alternating magnetic field environment, magnetic field intensity is 0.1mT-5mT, and field frequency is 2Hz-30Hz.
In step (4), the condition of described Subcritical Water Extraction is: water material mass ratio is 10-50:1, and extracting pressure is at 4MPa-25MPa, and extraction temperature is 100 DEG C-180 DEG C, and extraction time is 10min-90min.
In step (5), in the every 1L of described streptomycete liquid fermentation medium, extract residue II containing 0.2g-1g.Described streptomycete liquid fermentation medium forms with fermentation basal medium by extracting residue II, and described fermentation basal medium adopts the fermentation basal medium of this area routine, can adopt commercially available prod, and existing compound method also can be adopted to prepare.
General fermentation basal medium is made up of the component of following mass percent: glucose 1%-3%, K
2hPO
40.1%-0.2%, MgSO
4the water of 0.05%-0.1% and surplus.
The preparation method of described streptomycete liquid fermentation medium, comprising: 0.2g-1g is extracted residue II fermentation basal medium and be settled to 1L, mix, and pH value nature, obtains streptomycete liquid fermentation medium.
The access amount of described streptomycete seed liquor is the 3%-15% of streptomycete liquid fermentation medium volume.
Described mushroom Agaricus blazei Murill can adopt any one mushroom Agaricus blazei Murill existing, can adopt commercially available prod, such as: Brazilian mushroom (AgaricusblazeiMurill) ACCC50645 bacterial classification, is purchased from Chinese agriculture Microbiological Culture Collection administrative center.
Described streptomyces species can adopt any one streptomyces species existing, can adopt commercially available prod, such as: streptomycete (Streptomyces) ACCC40462 bacterial classification, is purchased from Chinese agriculture Microbiological Culture Collection administrative center.
Described liquid MRS culture medium adopts the liquid MRS culture medium of this area routine, can adopt commercially available prod, and existing compound method also can be adopted to prepare.Consisting of of general liquid MRS culture medium: peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, glucose 20.0g, sodium acetate 5.0g, diammonium hydrogen citrate 2.0g, Tween-80 1.0ml, dipotassium hydrogen phosphate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g and distilled water 1.0 liters.
Described PBS buffer solution and PBS, adopt the PBS buffer solution of this area routine, can adopt commercially available prod, existing compound method also can be adopted to prepare.General 1LPBS buffer solution: potassium dihydrogen phosphate 0.27g, sodium hydrogen phosphate 1.42g, sodium chloride 8g and potassium chloride 0.2g, adds the abundant stirring and dissolving of appropriate amount of deionized water, and then add concentrated hydrochloric acid and adjust pH to 7.2-7.4, last constant volume is to 1L.
The temperature that streptomyces species after mushroom Agaricus blazei Murill after described activation, activation is cultivated in liquid MRS culture medium is natural environment temperature, is preferably 20 DEG C-30 DEG C.1cm is accessed in general 1L liquid MRS culture medium
2-4cm
2streptomyces species bacterium block after mushroom Agaricus blazei Murill bacterium block after the activation of size, activation.The activation method of described mushroom Agaricus blazei Murill or streptomyces species adopts the actication of culture method of this area routine, comprise: the mushroom Agaricus blazei Murill of slant preservation or streptomyces species are inoculated on PDA plating medium, carry out activation culture, cultivation temperature 20 DEG C-30 DEG C, incubation time 3 days-25 days, obtains the streptomyces species after the mushroom Agaricus blazei Murill after activating or activation.The culture medium that described PDA plating medium adopts this area seed culture conventional, can adopt commercially available prod.Further preferably, described PDA plating medium: potato 200g, glucose 20g and agar 15g-20g, be settled to 1000mL with water.
The percentage of the volume of the thalline liquid that described inoculum concentration refers to an access and the ratio of culture volume.
Use after the present invention's culture medium used all needs sterilizing, the condition of sterilizing adopts the normal condition of this area, such as can at 120 DEG C-125 DEG C sterilizing 20min-30min.
The described composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, is preferably made up of 40 weight portion-55 weight portion radix tetrastigmes fermentation subcritical abstraction things I and 20 weight portion-35 weight portion streptomycete fermentation products III.
The composition that the present invention utilizes food medicine fungi fermentation radix tetrastigme to obtain has good hypolipemic function, can be used to prepare the health products or medicine with hypolipemic function.Described health products or medicine directly can be prepared by described composition and also can be prepared together with the auxiliary material of this area by described composition, product forms can be varied, such as comprise a soft capsule for the composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, its content is made up of 40 weight portion-55 weight portion radix tetrastigmes fermentation subcritical abstraction thing I, 20 weight portion-35 weight portion streptomycete fermentation products III and 10 weight portion-20 part by weight of vitamin E.
The bletilla striata (Bletillastriata) has another name called Lian Jicao, Gan Gen, Bai Gen, indocalamus orchid, Zhu Lan, purple orchid, purple a species of orchid, hundred large bamboo hats with a conical crown and broad brim, for the block root of the orchid family bletilla striata platymiscium bletilla striata Bletillastriata (Thunb.) Reichb.f. or narrow leaf bletilla striata B.ochraccaSchltr., underground block root oblateness or irregular rhombus, meat, yellow-white.Mainly be distributed in China, Japan and Upper Myanmar.The bletilla striata has medical value and Ornamental value widely.Medical value is mainly for astringing to arrest bleeding, detumescence and promoting granulation.
Brazilian mushroom (AgaricusblazeiMurill), have another name called Agricus blazei, originate in the South American region such as North American southern and Brazil, the composition of Brazilian mushroom comprises protein, fat, vitamin, ash content and carbohydrate etc., compared with other bacterium, the protein and sugar matter content of Brazilian mushroom is abundanter, and in fructification, polysaccharide body proteglycan matter compound content is more.
Streptomycete (Streptomyces) belongs to Gram positive actinomycetes, and substrate mycelium is not ruptured, and aerial hyphae physically well develops usually, forms the fibrillae of spores of long (sometimes short).Spore can not move, and epitheca often has the shape jewelrys such as wart, thorn or hair.
The present invention's raw material used all can adopt commercially available prod.
Beneficial effect of the present invention:
1, when the present invention carries out radix tetrastigme solid state fermentation, the direct Brazilian mushroom liquid seeds liquid adopted through hatching process containing liquid MRS culture medium two step of cholate, and the bletilla striata is added in culture medium, effectively the nutritional labeling in radix tetrastigme and active ingredient can be carried out biodegradation and utilization, effectively the stimulating component in radix tetrastigme be removed and conversion, contribute to the content and the activity that improve active component in final products.In step (4) and step (5), radix tetrastigme product by solid-state fermentation is carried out Subcritical Water Extraction, make that the activity of active ingredient in extract is at utmost retained, content is greatly improved; Again using the residue after extraction as the main culture medium of streptomycete liquid fermentation, and adopt the streptomycete seed liquor through hatching process containing liquid MRS culture medium two step of NaCl, recycled by integration, radix tetrastigme resource is fully utilized, further increases content and the activity of active component in final products.
2, the inventive method raw material sources are natural, quality controllable, and environmentally safe is suitable for suitability for industrialized production.
3, to compare the reduction of T-CHOL in serum and content of triglyceride more remarkable for the radix tetrastigme tunning of the present invention's composition of utilizing food medicine fungi fermentation radix tetrastigme to obtain and common fermentation, Lemna paucicostata, the composition that the present invention utilizes food medicine fungi fermentation radix tetrastigme to obtain and soft gel products thereof are described, significantly can reduce the T-CHOL in serum and content of triglyceride, there is significant hypolipemic function, can be used to prepare the health products or medicine with hypolipemic function.
Detailed description of the invention
Below in conjunction with some embodiments, content of the present invention is illustrated further, but content of the present invention is not limited in the following examples.
Brazilian mushroom (AgaricusblazeiMurill) ACCC50645 bacterial classification, is purchased from Chinese agriculture Microbiological Culture Collection administrative center.
Streptomycete (Streptomyces) ACCC40462 bacterial classification, is purchased from Chinese agriculture Microbiological Culture Collection administrative center.
Cholate: No. 3 cholate, i.e. sodium taurocholate, cholic acid-NaTDC salt mixture (in the permanent industry in Beijing chemical industry far away).
Embodiment 1
One, material prepares
PDA plating medium: potato 200g, glucose 20g and agar 15g, be settled to 1000ml with water, natural pH, sterilizing 20min at 121 DEG C.
The streptomyces species of the mushroom Agaricus blazei Murill of slant preservation, slant preservation is inoculated into respectively on PDA plating medium, carry out activation culture, cultivation temperature 22 DEG C, incubation time 10 days, obtains the streptomyces species after the mushroom Agaricus blazei Murill after activating, activation respectively.
Consisting of of liquid MRS culture medium: peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, glucose 20.0g, sodium acetate 5.0g, diammonium hydrogen citrate 2.0g, Tween-80 1.0ml, dipotassium hydrogen phosphate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g and distilled water 1.0 liters.
Fermentation basal medium is made up of the component of following mass percent: glucose 1%, K
2hPO
40.1%, MgSO
40.05% and the water of surplus.
1LPBS buffer solution: potassium dihydrogen phosphate 0.27g, sodium hydrogen phosphate 1.42g, sodium chloride 8g and potassium chloride 0.2g, adds the abundant stirring and dissolving of appropriate amount of deionized water, and then add concentrated hydrochloric acid and adjust pH to 7.4, last constant volume is to 1L.
Two, the preparation of radix tetrastigme fermented product
(1) 2cm is got
2mushroom Agaricus blazei Murill after size activation is inoculated in 1L liquid MRS culture medium to be cultivated 5 days in 25 DEG C, and nutrient solution collects the first thalline through the centrifugal 10min of 6000rpm; First thalline is suspended in cholate mass percentage be 0.2% containing cholate liquid MRS culture medium in hatch 10h in 25 DEG C, collect the second thalline through the centrifugal 10min of 6000rpm; By the second thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the second thalline liquid, by the second thalline liquid by 5% inoculum concentration be inoculated into cholate mass percentage be 0.2% containing cholate liquid MRS culture medium in cultivate 5 days again in 22 DEG C, obtain Brazilian mushroom liquid seeds liquid;
Wherein, the first thalline is 1:1 with the volume ratio of the liquid MRS culture medium containing cholate for first thalline that suspends, and the second thalline is 1:1 with the volume ratio for the liquid MRS culture medium of second thalline that suspends.
(2) 1cm is got
2streptomyces species after size activation is inoculated in 1L liquid MRS culture medium cultivates 15h in 25 DEG C, nutrient solution collects the 3rd thalline through the centrifugal 10min of 6000rpm, 3rd thalline is suspended in NaCl mass percentage be 5% hatch 8h containing in the liquid MRS culture medium of NaCl in 25 DEG C, collect the 4th thalline through the centrifugal 10min of 6000rpm; By the 4th thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the 4th thalline liquid, by the 4th thalline liquid by 3% inoculum concentration be inoculated into NaCl mass percentage be 5% containing NaCl liquid MRS culture medium in 25 DEG C cultivate 15h, obtain streptomycete seed liquor;
Wherein, the 3rd thalline is 1:1 with the volume ratio of the liquid MRS culture medium containing NaCl for the 3rd thalline that suspends, and the 4th thalline is 1:1 with the volume ratio for the liquid MRS culture medium of the 4th thalline that suspends.
(3) Brazilian mushroom liquid seeds liquid is accessed in radix tetrastigme solid-state fermentation culture medium by the access amount accounting for radix tetrastigme solid-state fermentation culture medium volume 15%, cultivate 10 days at 25 DEG C, then be placed in the alternating magnetic field environment that magnetic field intensity is 3mT, field frequency is 15Hz and cultivate 20 days again, cultured products obtains radix tetrastigme product by solid-state fermentation after vacuum freeze drying, pulverizing;
The preparation of radix tetrastigme solid-state fermentation culture medium comprises: fresh radix tetrastigme block root, bletilla striata block root are rinsed well under running water, dry, cutting is 1cm-2cm lengthy motion picture shape or fritter; Again 8g radix tetrastigme block root and 2g bletilla striata block root fermentation basal medium are settled to 1L, mix, pH value nature, obtains radix tetrastigme solid-state fermentation culture medium.
(4) radix tetrastigme product by solid-state fermentation was pulverized 40 mesh sieves, through Subcritical Water Extraction, the condition of Subcritical Water Extraction is: water material mass ratio is 30:1, extracting pressure is at 15MPa, extraction temperature is 150 DEG C, extraction time is 60min, and obtain radix tetrastigme fermentation subcritical abstraction thing I, after extraction, remaining residue is extracted residue II after vacuum freeze drying.
(5) streptomycete seed liquor is accessed in streptomycete liquid fermentation medium by the access amount accounting for streptomycete liquid fermentation medium volume 10%, cultivate 60h at 25 DEG C, obtain streptomycete fermentation product III through the centrifugal 10min of 6000rpm;
The preparation of streptomycete liquid fermentation medium comprises: 0.6g is extracted residue II fermentation basal medium and be settled to 1L, mix, and pH value nature, obtains streptomycete liquid fermentation medium.
The composition utilizing food medicine fungi fermentation radix tetrastigme to obtain is made up of 40 weight portion radix tetrastigmes fermentation subcritical abstraction things I and 20 weight portion streptomycete fermentation products III.
Three, the preparation of soft gel products: the filler of soft capsule is made up of 40 weight portion radix tetrastigmes fermentation subcritical abstraction thing I, 20 weight portion streptomycete fermentation products III and 10 part by weight of vitamin E.
Embodiment 2
One, material prepares
PDA plating medium: potato 200g, glucose 20g and agar 20g, be settled to 1000ml with water, natural pH, sterilizing 20min at 121 DEG C.
The streptomyces species of the mushroom Agaricus blazei Murill of slant preservation, slant preservation is inoculated into respectively on PDA plating medium, carry out activation culture, cultivation temperature 20 DEG C, incubation time 25 days, obtains the streptomyces species after the mushroom Agaricus blazei Murill after activating, activation respectively.
Fermentation basal medium is made up of the component of following mass percent: glucose 3%, K
2hPO
40.2%, MgSO
40.1% and the water of surplus.
Liquid MRS culture medium and PBS buffer solution are with embodiment 1.
Two, the preparation of radix tetrastigme fermented product
(1) 1cm is got
2mushroom Agaricus blazei Murill after size activation is inoculated in 1L liquid MRS culture medium to be cultivated 8 days in 20 DEG C, and nutrient solution collects the first thalline through the centrifugal 10min of 6000rpm; First thalline is suspended in cholate mass percentage be 0.05% containing cholate liquid MRS culture medium in hatch 20h in 18 DEG C, collect the second thalline through the centrifugal 10min of 6000rpm; By the second thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the second thalline liquid, by the second thalline liquid by 5% inoculum concentration be inoculated into cholate mass percentage be 0.05% containing cholate liquid MRS culture medium in cultivate 10 days again in 18 DEG C, obtain Brazilian mushroom liquid seeds liquid;
Wherein, the first thalline is 1:2 with the volume ratio of the liquid MRS culture medium containing cholate for first thalline that suspends, and the second thalline is 1:2 with the volume ratio for the liquid MRS culture medium of second thalline that suspends.
(2) 4cm is got
2streptomyces species after size activation is inoculated in 1L liquid MRS culture medium cultivates 3h in 30 DEG C, nutrient solution collects the 3rd thalline through the centrifugal 10min of 6000rpm, 3rd thalline is suspended in NaCl mass percentage be 8% hatch 15h containing in the liquid MRS culture medium of NaCl in 20 DEG C, collect the 4th thalline through the centrifugal 10min of 6000rpm; By the 4th thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the 4th thalline liquid, by the 4th thalline liquid by 3% inoculum concentration be inoculated into NaCl mass percentage be 8% containing NaCl liquid MRS culture medium in 20 DEG C cultivate 30h, obtain streptomycete seed liquor;
Wherein, the 3rd thalline is 1:3 with the volume ratio of the liquid MRS culture medium containing NaCl for the 3rd thalline that suspends, and the 4th thalline is 1:3 with the volume ratio for the liquid MRS culture medium of the 4th thalline that suspends.
(3) Brazilian mushroom liquid seeds liquid is accessed in radix tetrastigme solid-state fermentation culture medium by the access amount accounting for radix tetrastigme solid-state fermentation culture medium volume 25%, cultivate 20 days at 18 DEG C, then be placed in the alternating magnetic field environment that magnetic field intensity is 0.1mT, field frequency is 2Hz and cultivate 30 days again, cultured products obtains radix tetrastigme product by solid-state fermentation after vacuum freeze drying, pulverizing;
The preparation of radix tetrastigme solid-state fermentation culture medium comprises: fresh radix tetrastigme block root, bletilla striata block root are rinsed well under running water, dry, cutting is 1cm-2cm lengthy motion picture shape or fritter; Again 9.5g radix tetrastigme block root and 3.5g bletilla striata block root fermentation basal medium are settled to 1L, mix, pH value nature, obtains radix tetrastigme solid-state fermentation culture medium.
(4) radix tetrastigme product by solid-state fermentation was pulverized 40 mesh sieves, through Subcritical Water Extraction, the condition of Subcritical Water Extraction is: water material mass ratio is 50:1, extracting pressure is at 25MPa, extraction temperature is 180 DEG C, extraction time is 10min, and obtain radix tetrastigme fermentation subcritical abstraction thing I, after extraction, remaining residue is extracted residue II after vacuum freeze drying.
(5) streptomycete seed liquor is accessed in streptomycete liquid fermentation medium by the access amount accounting for streptomycete liquid fermentation medium volume 15%, cultivate 120h at 20 DEG C, obtain streptomycete fermentation product III through the centrifugal 10min of 6000rpm;
The preparation of streptomycete liquid fermentation medium comprises: 1g is extracted residue II fermentation basal medium and be settled to 1L, mix, and pH value nature, obtains streptomycete liquid fermentation medium.
The composition utilizing food medicine fungi fermentation radix tetrastigme to obtain is made up of 55 weight portion radix tetrastigmes fermentation subcritical abstraction things I and 35 weight portion streptomycete fermentation products III.
Three, the preparation of soft gel products: the filler of soft capsule is made up of 55 weight portion radix tetrastigmes fermentation subcritical abstraction thing I, 35 weight portion streptomycete fermentation products III and 20 part by weight of vitamin E.
Embodiment 3
One, material prepares
PDA plating medium, liquid MRS culture medium and PBS buffer solution are with embodiment 1.
The streptomyces species of the mushroom Agaricus blazei Murill of slant preservation, slant preservation is inoculated into respectively on PDA plating medium, carry out activation culture, cultivation temperature 30 DEG C, incubation time 3 days, obtains the streptomyces species after the mushroom Agaricus blazei Murill after activating, activation respectively.
Fermentation basal medium is made up of the component of following mass percent: glucose 2%, K
2hPO
40.15%, MgSO
40.08% and the water of surplus.
Two, the preparation of radix tetrastigme fermented product
(1) 4cm is got
2mushroom Agaricus blazei Murill after size activation is inoculated in 1L liquid MRS culture medium to be cultivated 2 days in 30 DEG C, and nutrient solution collects the first thalline through the centrifugal 10min of 6000rpm; First thalline is suspended in cholate mass percentage be 0.5% containing cholate liquid MRS culture medium in hatch 1h in 30 DEG C, collect the second thalline through the centrifugal 10min of 6000rpm; By the second thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the second thalline liquid, by the second thalline liquid by 5% inoculum concentration be inoculated into cholate mass percentage be 0.5% containing cholate liquid MRS culture medium in cultivate 1 day again in 30 DEG C, obtain Brazilian mushroom liquid seeds liquid;
Wherein, the first thalline is 1:3 with the volume ratio of the liquid MRS culture medium containing cholate for first thalline that suspends, and the second thalline is 1:3 with the volume ratio for the liquid MRS culture medium of second thalline that suspends.
(2) 2cm is got
2streptomyces species after size activation is inoculated in 1L liquid MRS culture medium cultivates 30h in 20 DEG C, nutrient solution collects the 3rd thalline through the centrifugal 10min of 6000rpm, 3rd thalline is suspended in NaCl mass percentage be 0.5% hatch 0.5h containing in the liquid MRS culture medium of NaCl in 30 DEG C, collect the 4th thalline through the centrifugal 10min of 6000rpm; By the 4th thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the 4th thalline liquid, by the 4th thalline liquid by 3% inoculum concentration be inoculated into NaCl mass percentage be 0.5% containing NaCl liquid MRS culture medium in 30 DEG C cultivate 3h, obtain streptomycete seed liquor;
Wherein, the 3rd thalline is 1:2 with the volume ratio of the liquid MRS culture medium containing NaCl for the 3rd thalline that suspends, and the 4th thalline is 1:2 with the volume ratio for the liquid MRS culture medium of the 4th thalline that suspends.
(3) Brazilian mushroom liquid seeds liquid is accessed in radix tetrastigme solid-state fermentation culture medium by the access amount accounting for radix tetrastigme solid-state fermentation culture medium volume 5%, cultivate 2 days at 30 DEG C, then be placed in the alternating magnetic field environment that magnetic field intensity is 5mT, field frequency is 30Hz and cultivate 5 days again, cultured products obtains radix tetrastigme product by solid-state fermentation after vacuum freeze drying, pulverizing;
The preparation of radix tetrastigme solid-state fermentation culture medium comprises: fresh radix tetrastigme block root, bletilla striata block root are rinsed well under running water, dry, cutting is 1cm-2cm lengthy motion picture shape or fritter; Again 6.5g radix tetrastigme block root and 0.5g bletilla striata block root fermentation basal medium are settled to 1L, mix, pH value nature, obtains radix tetrastigme solid-state fermentation culture medium.
(4) radix tetrastigme product by solid-state fermentation was pulverized 40 mesh sieves, through Subcritical Water Extraction, the condition of Subcritical Water Extraction is: water material mass ratio is 10:1, extracting pressure is at 4MPa, extraction temperature is 100 DEG C, extraction time is 90min, and obtain radix tetrastigme fermentation subcritical abstraction thing I, after extraction, remaining residue is extracted residue II after vacuum freeze drying.
(5) streptomycete seed liquor is accessed in streptomycete liquid fermentation medium by the access amount accounting for streptomycete liquid fermentation medium volume 3%, cultivate 10h at 30 DEG C, obtain streptomycete fermentation product III through the centrifugal 10min of 6000rpm;
The preparation of streptomycete liquid fermentation medium comprises: 0.2g is extracted residue II fermentation basal medium and be settled to 1L, mix, and pH value nature, obtains streptomycete liquid fermentation medium.
The composition utilizing food medicine fungi fermentation radix tetrastigme to obtain is made up of 50 weight portion radix tetrastigmes fermentation subcritical abstraction things I and 25 weight portion streptomycete fermentation products III.
Three, the preparation of soft gel products: the filler of soft capsule is made up of 50 weight portion radix tetrastigmes fermentation subcritical abstraction thing I, 25 weight portion streptomycete fermentation products III and 15 part by weight of vitamin E.
Comparative example 1
One, material prepares
Mushroom Agaricus blazei Murill after PDA plating medium, activation, liquid MRS culture medium and fermentation basal medium are all with embodiment 1.
Two, the preparation of radix tetrastigme fermented product
(1) 2cm is got
2mushroom Agaricus blazei Murill after size activation is inoculated in 1L liquid MRS culture medium to be cultivated 5 days in 25 DEG C, obtains Brazilian mushroom liquid seeds liquid.
(2) access in radix tetrastigme fermentation medium by Brazilian mushroom liquid seeds liquid by the access amount accounting for radix tetrastigme fermentation medium volume 15%, cultivate 10 days at 25 DEG C, cultured products obtains radix tetrastigme tunning after vacuum freeze drying, pulverizing;
The preparation of radix tetrastigme fermentation medium comprises: rinsed well under running water by fresh radix tetrastigme block root, dry, cutting is 1cm-2cm lengthy motion picture shape or fritter; Again 8g radix tetrastigme block root fermentation basal medium is settled to 1L, mixes, pH value nature, obtains radix tetrastigme fermentation medium.
Three, the preparation of soft gel products: the filler of soft capsule is made up of 40 weight portion radix tetrastigme tunnings and 10 part by weight of vitamin E.
Comparative example 2
Rinsed well under running water by fresh radix tetrastigme block root, dry, cutting is 1cm-2cm lengthy motion picture shape or fritter; Add water by feed liquid weight ratio 1:10,10h is extracted under fluidized state, filter to obtain first-time filtrate and a filter residue, a filter residue is added water by feed liquid weight ratio 1:5,8h is extracted under fluidized state, filter to obtain secondary filtrate, merge first-time filtrate and secondary filtrate, after freeze drying, pulverizing, obtain Lemna paucicostata.
The preparation of soft gel products: the filler of soft capsule is made up of 40 weight portion Lemna paucicostata and 10 part by weight of vitamin E.
The mensuration of active component
(1) measurement of the polysaccharide content
Take appropriate amount of sample dry powder, cross 40 orders after pulverizing, be placed in beaker, add distilled water by solid-liquid ratio 1:10 (mass ratio), 90 DEG C of lixiviate 3h, extract twice, filtration under diminished pressure collects filtrate, reduced pressure concentration at 50 DEG C, concentrate adds 4 times of volume absolute ethyl alcohols, alcohol precipitation overnight, after 3000r/min, 20min are centrifugal, precipitation is Thick many candies.Precipitation distilled water is settled to 50mL, and measure polysaccharide in fermentation liquid content with phend-sulphuric acid, replication is averaged for 3 times, i.e. polyoses content.
(2) mensuration of flavones content
Be dried to the rutin standard items 10.0mg of constant weight under accurately taking 120 DEG C of conditions, dissolve with the ethanol water of mass percentage concentration 70% and be settled to 100mL, being made into 0.1mg/mL standard liquid.Get respectively standard liquid 0.0,1.0,2.0,3.0,4.0,5.0mL in 10mL volumetric flask, after adding mass percentage concentration 70% ethanol water to 5.0mL respectively, add the NaNO of 0.3mL mass percentage concentration 5%
2the aqueous solution, shakes up and places 6min; Add the Al (NO of 0.4mL mass percentage concentration 10% again
3)
3the aqueous solution, shakes up and places 6min; Add the 4.0mL mass percentage concentration 4%NaOH aqueous solution again, mass percentage concentration 60% ethanol water is settled to scale, shake up and place 15min, be cooled to the absorbance at 510nm place bioassay standard product after room temperature, with standard solution mass concentration C (mg/mL) for abscissa, absorbance A is ordinate drawing standard curve, obtains regression equation.
Take appropriate amount of sample dry powder, cross 40 orders after pulverizing, first add the ethanol water of mass percentage concentration 70% by solid-liquid ratio 1:30 (mass ratio), at 70 DEG C, extract 2h, be settled to 50ml after extract centrifugal filtration, therefrom get 1ml sample and carry out assay.
The each embodiment of table 1 and comparative example main component testing result
Note: compare with comparative example 1, comparative example 2, Δ Δ: P<0.01.
Shown by the data of table 1, the composition that the present invention utilizes food medicine fungi fermentation radix tetrastigme to obtain, significantly improve the content of polysaccharide in product, flavones isoreactivity composition simultaneously.Compared with comparative example 1 conventional culture methods, in the radix tetrastigme composition adopting the inventive method to obtain, polyoses content improves 68.2%-71.1%, and flavones content improves 80.6%-92.2%.
Auxiliary antilipemic is tested
(1) animal used as test grouping
Mouse 24-28g, male and female half and half, according to the horizontal random packet of TC after adaptability grows 1 week, often organize 10.By the effect measuring serum total cholesterol (TC), triglycerides (TG) level judges reducing blood lipid.
(2) medication and dosage
High lipid food is the KK mouse material of Fukang bio tech ltd of China, Beijing, crude fat mass percentage 16.15%, gives the given the test agent (the radix tetrastigme tunning in the composition namely in embodiment 1-3, comparative example 1 and the Lemna paucicostata in comparative example 1) of various dose while giving high lipid food.5 process are established in experiment, i.e. high, medium and low 3 dosage groups (8.0ml/kg, 16.0ml/kg, 24.0ml/kg), Basal control group and high fat control group, high fat control group and Basal control group are all with distilled water gavage, and give high lipid food and basal feed (purchased from medical animal field, Guangdong Province) respectively, synchronously carry out with dosage group.Each dosage group gavage every day 1 soft capsule sample, feeds 30 days continuously, while giving given the test agent, feed high lipid food, ad lib and drinking-water.
(3) content and method is measured
This experiment is preventive administration, the given the test agent of various dose is given while giving high lipid food, fasting 16 hours are terminated in experiment, measure described in kit (COD-PAP method) and the middle description of triglyceride determination kit (GPO-PAP method) according to T-CHOL, survey serum total cholesterol (TC), triglycerides (TG) level respectively.Testing result is in table 2.
Table 2 soft capsule sample is on the impact (X ± S, n=10) of mouse TC and TG
△ represents and compares P < 0.05, significant difference with high fat control group; * represent and compare P < 0.05, significant difference with comparative example 1 matched doses group.
Table 2 data show, the composition adopting the present invention to utilize food medicine fungi fermentation radix tetrastigme to obtain and the radix tetrastigme tunning of comparative example 1 common fermentation, the reduction that comparative example 2 Lemna paucicostata compares T-CHOL in serum and content of triglyceride is more remarkable, serum total cholesterol (TC) falls in the composition that the present invention utilizes food medicine fungi fermentation radix tetrastigme to obtain, the effect of triglycerides (TG) obviously will be better than the radix tetrastigme tunning of comparative example 1 common fermentation, comparative example 2 Lemna paucicostata, the composition that the present invention utilizes food medicine fungi fermentation radix tetrastigme to obtain and soft gel products thereof are described, significantly can reduce the T-CHOL in serum and content of triglyceride, there is significant hypolipemic function.
In the scope that formula of the present invention and preparation method limit, the change of each parameter does not affect blood fat reducing function and the preparation of composition that the present invention utilizes food medicine fungi fermentation radix tetrastigme to obtain and soft gel products thereof, the composition that therefore in formula of the present invention and preparation method, the combination of arbitrary parameter all can obtain utilizing food medicine fungi fermentation radix tetrastigme to obtain and soft gel products thereof.Do not repeat them here.
Claims (10)
1. a preparation method for the composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, is characterized in that, comprise step:
(1) mushroom Agaricus blazei Murill got after activation is inoculated in liquid MRS culture medium and cultivates 2 days-8 days, collected by centrifugation first thalline; First thalline is suspended in the liquid MRS culture medium containing cholate and hatches 1h-20h in 18 DEG C-30 DEG C, collected by centrifugation second thalline; By the second thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the second thalline liquid, second thalline liquid is inoculated in the liquid MRS culture medium containing cholate and cultivates 1 day-10 days again in 18 DEG C-30 DEG C, obtain Brazilian mushroom liquid seeds liquid;
(2) streptomyces species got after activation is inoculated in liquid MRS culture medium and cultivates 3h-30h, collected by centrifugation the 3rd thalline, is suspended in by the 3rd thalline in the liquid MRS culture medium containing NaCl and hatches 0.5h-15h in 20 DEG C-30 DEG C, collected by centrifugation the 4th thalline; By the 4th thalline after PBS buffer solution for cleaning is clean Eddy diffusion in liquid MRS culture medium, concussion shakes up and obtains the 4th thalline liquid, 4th thalline liquid is inoculated in the liquid MRS culture medium containing NaCl and cultivates 3h-30h in 20 DEG C-30 DEG C, obtain streptomycete seed liquor;
(3) by the Brazilian mushroom liquid seeds liquid of step (1) access radix tetrastigme solid-state fermentation culture medium, cultivate 2 days-20 days at 18 DEG C-30 DEG C, then be placed in alternating magnetic field environment and cultivate 5 days-30 days again, cultured products obtains radix tetrastigme product by solid-state fermentation after vacuum freeze drying, pulverizing; Containing radix tetrastigme and the bletilla striata in described radix tetrastigme solid-state fermentation culture medium;
(4) by the radix tetrastigme product by solid-state fermentation of step (3) through Subcritical Water Extraction, obtain radix tetrastigme fermentation subcritical abstraction thing I, after extraction, remaining residue is extracted residue II after vacuum freeze drying;
(5) by the streptomycete seed liquor of step (2) access streptomycete liquid fermentation medium, 10h-120h is cultivated at 20 DEG C-30 DEG C, centrifugal, obtain streptomycete fermentation product III, containing extraction residue II in described streptomycete liquid fermentation medium;
The described composition utilizing food medicine fungi fermentation radix tetrastigme to obtain is radix tetrastigme fermentation subcritical abstraction thing I and streptomycete fermentation product III.
2. preparation method according to claim 1, is characterized in that, in step (3), containing 6.5g-9.5g radix tetrastigme block root and 0.5g-3.5g bletilla striata block root in the every 1L of described radix tetrastigme solid-state fermentation culture medium.
3. preparation method according to claim 1, is characterized in that, in step (5), extracts residue II in the every 1L of described streptomycete liquid fermentation medium containing 0.2g-1g.
4. preparation method according to claim 1, is characterized in that, in step (1), described is 0.05%-0.5% containing the mass percentage of cholate in the liquid MRS culture medium of cholate;
In step (2), the mass percentage of the described liquid MRS NaCl in medium containing NaCl is 0.5%-8%.
5. preparation method according to claim 1, it is characterized in that, in step (1), described first thalline is 1:1-3 with the volume ratio of the liquid MRS culture medium containing cholate for first thalline that suspends, second thalline is 1:1-3 with the volume ratio for the liquid MRS culture medium of second thalline that suspends, and the inoculum concentration of the second thalline liquid is 5%;
In step (2), described 3rd thalline is 1:1-3 with the volume ratio of the liquid MRS culture medium containing NaCl for the 3rd thalline that suspends, 4th thalline is 1:1-3 with the volume ratio for the liquid MRS culture medium of the 4th thalline that suspends, and the inoculum concentration of the 4th thalline liquid is 3%.
6. preparation method according to claim 1, is characterized in that, in step (3), in described alternating magnetic field environment, magnetic field intensity is 0.1mT-5mT, and field frequency is 2Hz-30Hz;
In step (4), the condition of described Subcritical Water Extraction is: water material mass ratio is 10-50:1, and extracting pressure is at 4MPa-25MPa, and extraction temperature is 100 DEG C-180 DEG C, and extraction time is 10min-90min.
7. preparation method according to claim 1, is characterized in that, in step (3), the access amount of Brazilian mushroom liquid seeds liquid is the 5%-25% of radix tetrastigme solid-state fermentation culture medium volume;
In step (5), the access amount of described streptomycete seed liquor is the 3%-15% of streptomycete liquid fermentation medium volume.
8. the composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, is characterized in that, adopt the preparation method described in any one of claim 1-7 to prepare.
9. the composition utilizing food medicine fungi fermentation radix tetrastigme to obtain according to claim 8, it is characterized in that, described composition is made up of 40 weight portion-55 weight portion radix tetrastigmes fermentation subcritical abstraction things I and 20 weight portion-35 weight portion streptomycete fermentation products III.
10. an application for the composition utilizing food medicine fungi fermentation radix tetrastigme to obtain, is characterized in that, the application of composition in the health products or medicine of preparation reducing blood lipid that utilization food medicine fungi fermentation radix tetrastigme according to claim 8 or claim 9 obtains.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107259514A (en) * | 2017-05-02 | 2017-10-20 | 江苏大学 | A kind of method of White mushroom mushroom handle comprehensive utilization of resources |
| CN112029703A (en) * | 2020-06-05 | 2020-12-04 | 浙江农林大学 | Method for improving content of secondary metabolites based on coculture system of radix tetrastigme leaf suspension cells and endophytic fungi thereof |
| CN113350428A (en) * | 2021-06-16 | 2021-09-07 | 山东中医药大学 | Fermentation method of radix tetrastigme |
| CN114958620A (en) * | 2022-05-30 | 2022-08-30 | 杭州市农业科学研究院 | Phellinus igniarius mycelium liquid fermentation culture medium and fermentation culture method |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1704867A1 (en) * | 2005-03-18 | 2006-09-27 | Crossbeta Biosciences B.V. | Cross-beta structures on microbial organisms |
| CN101417000A (en) * | 2008-09-11 | 2009-04-29 | 宁波保税区欣诺生物技术有限公司 | New preparation method of Tetrastigma hemsleyanum Diels et Gilg extract |
| CN103829249A (en) * | 2014-03-18 | 2014-06-04 | 苏州科大微龙信息技术有限公司 | Enzyme for conditioning spleen and stomach damp-heat constitution and preparation method thereof |
| CN103964969A (en) * | 2014-05-28 | 2014-08-06 | 海峡（厦门）现代农业研究院有限公司 | Special culture substrate for traditional Chinese medicinal materials and application of special culture substrate |
| CN104082819A (en) * | 2014-07-04 | 2014-10-08 | 杭州雪域生物技术有限公司 | Edible and medical fungus functional solid beverage and preparation method thereof |
| CN104630116A (en) * | 2015-02-10 | 2015-05-20 | 中国科学院南海海洋研究所 | Streptomyces sp. and application thereof |
-
2015
- 2015-12-31 CN CN201511027688.3A patent/CN105559068B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1704867A1 (en) * | 2005-03-18 | 2006-09-27 | Crossbeta Biosciences B.V. | Cross-beta structures on microbial organisms |
| CN101417000A (en) * | 2008-09-11 | 2009-04-29 | 宁波保税区欣诺生物技术有限公司 | New preparation method of Tetrastigma hemsleyanum Diels et Gilg extract |
| CN103829249A (en) * | 2014-03-18 | 2014-06-04 | 苏州科大微龙信息技术有限公司 | Enzyme for conditioning spleen and stomach damp-heat constitution and preparation method thereof |
| CN103964969A (en) * | 2014-05-28 | 2014-08-06 | 海峡（厦门）现代农业研究院有限公司 | Special culture substrate for traditional Chinese medicinal materials and application of special culture substrate |
| CN104082819A (en) * | 2014-07-04 | 2014-10-08 | 杭州雪域生物技术有限公司 | Edible and medical fungus functional solid beverage and preparation method thereof |
| CN104630116A (en) * | 2015-02-10 | 2015-05-20 | 中国科学院南海海洋研究所 | Streptomyces sp. and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| DING GANG-QIANG，ETAL: "Toxicological Effects of the Extract of Tetrastigma hemsleyanum Diels et. Gilg on Hepatocellular Carcinoma Cell Line HepG_2 and Primary Rat Hepatocytes in vitro", 《ZHEJIANG JOURNAL OF PREVENTIVE MEDICINE》 * |
| 刘东，等: "中国特有植物三叶青化学成分的研究", 《中国中药杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107259514A (en) * | 2017-05-02 | 2017-10-20 | 江苏大学 | A kind of method of White mushroom mushroom handle comprehensive utilization of resources |
| CN112029703A (en) * | 2020-06-05 | 2020-12-04 | 浙江农林大学 | Method for improving content of secondary metabolites based on coculture system of radix tetrastigme leaf suspension cells and endophytic fungi thereof |
| CN113350428A (en) * | 2021-06-16 | 2021-09-07 | 山东中医药大学 | Fermentation method of radix tetrastigme |
| CN114958620A (en) * | 2022-05-30 | 2022-08-30 | 杭州市农业科学研究院 | Phellinus igniarius mycelium liquid fermentation culture medium and fermentation culture method |
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