CN105559068B - It is a kind of to utilize composition for eating the acquisition of medicine fungi fermentation radix tetrastigme and preparation method thereof - Google Patents
It is a kind of to utilize composition for eating the acquisition of medicine fungi fermentation radix tetrastigme and preparation method thereof Download PDFInfo
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- CN105559068B CN105559068B CN201511027688.3A CN201511027688A CN105559068B CN 105559068 B CN105559068 B CN 105559068B CN 201511027688 A CN201511027688 A CN 201511027688A CN 105559068 B CN105559068 B CN 105559068B
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- radix tetrastigme
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- C—CHEMISTRY; METALLURGY
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- 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
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Abstract
The invention discloses a kind of using composition for eating the acquisition of medicine fungi fermentation radix tetrastigme and preparation method thereof, said composition is radix tetrastigme fermentation subcritical abstraction thing I and streptomycete fermentation product III, has good hypolipemic function.Its preparation includes：Brazilian mushroom liquid seeds liquid is accessed in the radix tetrastigme solid-state fermentation culture medium containing radix tetrastigme and the bletilla striata and cultivated, it is subsequently placed in alternating magnetic field environment and is further cultured for, radix tetrastigme product by solid-state fermentation is obtained, then through Subcritical Water Extraction, obtains radix tetrastigme fermentation subcritical abstraction thing I；It will be cultivated in streptomycete liquid fermentation medium of the streptomycete seed liquor access containing extraction residue II, obtain streptomycete fermentation product III；Nutritional ingredient in radix tetrastigme and active ingredient effectively can be carried out content and activity biodegradable and that utilize, be favorably improved active component in final products by this method.
Description
Technical field
The present invention relates to the processing and utilization field of radix tetrastigme, and in particular to one kind is obtained using medicine fungi fermentation radix tetrastigme is eaten
Composition and preparation method thereof.
Background technology
Radix tetrastigme (Tetrastigma hemsleyanum Diels et Gilg) is Vitaceae Tetrastigma plant, not
Name：Gold thread hoist, stone mouse, hemsley rockvine root, category Rhamnales, Vitaceae herbaceous species.Scientific name：Tetratigma hemsleyanum, it is Zhejiang, good fortune
The regional tradition such as build and commonly use green herbal medicine, it is best with the medicinal effects of underground root tuber and fruit.Treated with radix tetrastigme Zhejiang is among the people
Tumour is more remote, and radix tetrastigme is region medicinal material, extremely rare, be distributed in Zhejiang, Jiangxi, Fujian, Anhui, Guangdong, Guangxi,
The ground such as Yunnan, radix tetrastigme has the function that radiating, qi-regulating, promoting blood circulation, tonifying spleen, removing toxic substances, wind-dispelling, anti-inflammatory, among the people to be used for treating children
High fever, pharyngo-laryngitis chronica, tonsillitis, hepatitis, pneumonia, gastritis, scrofula and snake bite etc. have special effect.Modern pharmacological research
Show that radix tetrastigme has anti-inflammatory, analgesia and antipyretic, antitumor, the antiviral, effect such as regulation is immune.
At present, the active component in radix tetrastigme is obtained using traditional extracting and developing method more, such as Chinese patent
The Chinese medicine radix tetrastigme for being used to prevent and treat tumour disclosed in ZL200510050649.5, it is original using by radix tetrastigme
The water and/or alcohol extracting thing of material make active component.Overcritical dioxy disclosed in Chinese patent application CN201110264176.4
Change the method for carbon extraction radix tetrastigme general flavone, by radix tetrastigme pulverizing medicinal materials 60-80 mesh, add supercritical carbon dioxide extracting tank,
Liquid carbon dioxide and entrainer are passed through, under the conditions of pressure 20-30MPa, 40-50 DEG C of temperature, extracts 2-3 hours, separator
Middle decompression parsing extract, reclaim reagent, dry radix tetrastigme general flavone.Chinese patent ZL200810120750.7 discloses one
The technique of kind high efficiency extraction flavone compound from radix tetrastigme, its technological process are:Raw material radix tetrastigme, air-dry, shred, add
Water extracts in ultrasonic extractor, extract solution is concentrated into certain volume, then carry out adsorption bleaching with activated carbon.Followed by
Cavitation suspension extraction separator, is first extracted with ethyl acetate, and reclaims ethyl acetate, aqueous solution part is extracted with chloroform, returns
Chloroform is received, by aqueous solution partial concentration to without organic reagent taste, by macroporous absorbent resin, is washed successively with water and 50% ethanol
It is de-, 50% ethanol eluate is collected, recycling design obtains Lemna paucicostata to doing.
Containing the nutritional ingredient such as polysaccharide, flavones, polyphenol isoreactivity composition and starch, protein in radix tetrastigme, traditional carries
Take, separate in its active component technique, starch, protein, stimulating component etc. extract together into branch, add in separation
Processing step, cause the active component content in final products it is relatively low, activity it is relatively low.
The content of the invention
The invention provides a kind of using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, active component in said composition
Content and activity it is high, there is good hypolipemic function.
Present invention also offers a kind of preparation method using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, using true
Bacterium fermentation radix tetrastigme, is favorably improved the content and activity of active component in final products, obtains the hair with hypolipemic function
Ferment composition.
It is a discovery of the invention that according to radix tetrastigme nutritional ingredient and active ingredient feature, pass through the specific fungi fermentation of the present invention
And specific technique, and the bletilla striata is added into culture medium, effectively the nutritional ingredient in radix tetrastigme and active ingredient can be entered
Row is biodegradable and utilizes, effectively the stimulating component in radix tetrastigme is removed and converted, and radix tetrastigme resource is obtained fully
Utilization, be favorably improved the content and activity of active component in final products.
The technical solution adopted by the present invention is：
A kind of preparation method using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, including step：
(1) take the mushroom Agaricus blazei Murill after activation to be inoculated into liquid MRS culture mediums and cultivate -8 days 2 days, be collected by centrifugation the
One thalline；First thalline is suspended in the liquid MRS culture mediums containing cholate in 18 DEG C of -30 DEG C of incubation 1h-20h, is collected by centrifugation
Second thalline；Second thalline is resuspended in liquid MRS culture mediums after PBS cleans up, concussion shakes up
To the second thalline liquid, the second thalline liquid is inoculated into the liquid MRS culture mediums containing cholate and is further cultured for 1 day -10 in 18 DEG C -30 DEG C
My god, obtain Brazilian mushroom liquid seeds liquid；
(2) take the streptomyces species after activation to be inoculated into liquid MRS culture mediums and cultivate 3h-30h, the 3rd bacterium is collected by centrifugation
Body, the 3rd thalline is suspended in the liquid MRS culture mediums containing NaCl in 20 DEG C of -30 DEG C of incubation 0.5h-15h, is collected by centrifugation the
Four thalline；4th thalline is resuspended in after PBS cleans up in liquid MRS culture mediums, concussion shakes up to obtain
4th thalline liquid, the 4th thalline liquid is inoculated into the liquid MRS culture mediums containing NaCl in 20 DEG C of -30 DEG C of culture 3h-30h, obtained
To streptomycete seed liquor；
(3) the Brazilian mushroom liquid seeds liquid of step (1) is accessed in radix tetrastigme solid-state fermentation culture medium, at 18 DEG C -30
DEG C culture -20 days 2 days, be subsequently placed in alternating magnetic field environment and be further cultured for -30 days 5 days, cultured products are through vacuum freeze drying, powder
Radix tetrastigme product by solid-state fermentation is obtained after broken；Contain radix tetrastigme and the bletilla striata in the radix tetrastigme solid-state fermentation culture medium；
(4) the radix tetrastigme product by solid-state fermentation of step (3) is obtained into the subcritical extraction of radix tetrastigme fermentation through Subcritical Water Extraction
Thing I is taken, remaining residue obtains extracting residue II after vacuum freeze drying after extraction.
(5) by the streptomycete seed liquor access streptomycete liquid fermentation medium of step (2), cultivated at 20 DEG C -30 DEG C
10h-120h, centrifugation, obtains streptomycete fermentation product III；Contain extraction residue in the streptomycete liquid fermentation medium
Ⅱ；
The composition that the acquisition of medicine fungi fermentation radix tetrastigme is eaten in the utilization is radix tetrastigme fermentation subcritical abstraction thing I and strepto-
Bacterium tunning III.
When the present invention carries out radix tetrastigme solid state fermentation, directly using through at the step incubation of liquid MRS culture mediums two containing cholate
The Brazilian mushroom liquid seeds liquid of reason, and the bletilla striata is added into culture medium, can effectively by the nutritional ingredient in radix tetrastigme and
Active ingredient is carried out biodegradable and utilized, effectively the stimulating component in radix tetrastigme is removed and converted, and is favorably improved
The content and activity of active component in final products.In step (4) and step (5), radix tetrastigme product by solid-state fermentation is carried out sub-
Critical extracts, and enables that the activity of active ingredient in extract at utmost retains, content is greatly improved；Again will extraction
The culture medium main as streptomycete liquid fermentation of residue afterwards, and be incubated using through the step of liquid MRS culture mediums two containing NaCl
The streptomycete seed liquor of processing, is recycled by integration, radix tetrastigme resource is fully utilized, is further increased
The content and activity of active component in final products.
In order to reach more preferable effect, preferably：
In step (1), the weight/mass percentage composition of cholate is 0.05%- in the liquid MRS culture mediums containing cholate
0.5%.
The volume ratio of first thalline and the liquid MRS culture mediums containing cholate for first thalline that suspends is 1:1-3,
The volume ratio of second thalline and the liquid MRS culture mediums for second thalline that suspends is 1:1-3, the inoculum concentration of the second thalline liquid are
5%.
In step (2), the weight/mass percentage composition of the liquid MRS NaCl in medium containing NaCl is 0.5%-8%.
The volume ratio of 3rd thalline and the liquid MRS culture mediums containing NaCl for the 3rd thalline that suspends is 1:1-3,
The volume ratio of 4th thalline and the liquid MRS culture mediums for the 4th thalline that suspends is 1:1-3, the inoculum concentration of the 4th thalline liquid are
3%.
In step (3), the radix tetrastigme uses radix tetrastigme root tuber；The bletilla striata uses bletilla striata root tuber.The radix tetrastigme is consolidated
Contain 6.5g-9.5g radix tetrastigmes root tuber and 0.5g-3.5g bletilla striata root tubers in the every 1L of state fermentation medium.The radix tetrastigme solid-state hair
Ferment culture medium is made up of radix tetrastigme root tuber, bletilla striata root tuber and fermentation basal medium, and the fermentation basal medium uses ability
The conventional fermentation basal medium in domain, can use commercially available prod, can also use existing compound method to prepare.
The preparation method of the radix tetrastigme solid-state fermentation culture medium, including：By fresh radix tetrastigme root tuber, bletilla striata root tuber certainly
Rinse well, dry under water, cutting is the long sheets of 1cm-2cm or fritter；Again by 6.5g-9.5g radix tetrastigmes root tuber and 0.5g-
3.5g bletilla striatas root tuber is settled to 1L with fermentation basal medium, is well mixed, pH value is naturally, obtain radix tetrastigme solid state fermentation culture
Base.
The access amount of the Brazilian mushroom liquid seeds liquid is the 5%-25% of radix tetrastigme solid-state fermentation culture medium volume.
Fungi solid-state fermentation technology generally existing fermentation period is long at present, mycelial growth sprouting is slow, feature secondary metabolism generation
Thank to the problems such as product assay is low.Present invention discover that alternating magnetic field treatment technology is applied in solid state fermentation, pass through specific culture
The extraneous factor effective stimulus such as the proper treatment in period, improve growth metabolism of the hypha,hyphae on radix tetrastigme culture base-material, contracting
Short fermentation time, effectively facilitate the generation of secondary metabolite.Magnetic field intensity is 0.1mT-5mT in the alternating magnetic field environment,
Field frequency is 2Hz-30Hz.
In step (4), the condition of the Subcritical Water Extraction is：Water material mass ratio is 10-50:1, extracting pressure exists
4MPa-25MPa, extraction temperature are 100 DEG C -180 DEG C, extraction time 10min-90min.
In step (5), 0.2g-1g extraction residues II are contained in the every 1L of the streptomycete liquid fermentation medium.It is described
Streptomycete liquid fermentation medium is made up of extraction residue II and fermentation basal medium, and described fermentation basal medium is adopted
With the conventional fermentation basal medium in this area, commercially available prod can be used, can also use existing compound method to prepare.
General fermentation basal medium is made up of the component of following mass percent：Glucose 1%-3%, K2HPO4
0.1%-0.2%, MgSO40.05%-0.1% and surplus water.
The preparation method of the streptomycete liquid fermentation medium, including：By 0.2g-1g extraction residues II fermentation base
Basal culture medium is settled to 1L, is well mixed, pH value is naturally, obtain streptomycete liquid fermentation medium.
The access amount of the streptomycete seed liquor is the 3%-15% of streptomycete liquid fermentation medium volume.
The mushroom Agaricus blazei Murill can use any one existing mushroom Agaricus blazei Murill, can use commercially available prod, such as：Bar
Western mushroom (Agaricus blazei Murill) ACCC50645 strains, it is purchased from the management of Chinese agriculture Microbiological Culture Collection
The heart.
The streptomyces species can use any one existing streptomyces species, can use commercially available prod, such as：Streptomycete
(Streptomyces) ACCC40462 strains, it is purchased from Chinese agriculture Microbiological Culture Collection administrative center.
The liquid MRS culture mediums can be used commercially available prod, can also be adopted using the conventional liquid MRS culture mediums in this area
Prepared with existing compound method.The composition of general liquid MRS culture mediums is：Peptone 10.0g, beef extract 10.0g, yeast leaching
Cream 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 1.0 liters of distilled water.
The PBS is phosphate buffered saline solution, using the conventional PBS in this area, can use commercially available production
Product, can also existing compound method be used to prepare.General 1L PBSs：Potassium dihydrogen phosphate 0.27g, disodium hydrogen phosphate 1.42g,
Sodium chloride 8g and potassium chloride 0.2g, adds appropriate amount of deionized water to be sufficiently stirred dissolving, then adds concentrated hydrochloric acid and adjusts pH to 7.2-7.4,
Last constant volume is to 1L.
The temperature that the streptomyces species after mushroom Agaricus blazei Murill, activation after the activation are cultivated in liquid MRS culture mediums
Degree is natural environment temperature, preferably 20 DEG C -30 DEG C.1cm is accessed in general 1L liquid MRS culture mediums2-4cm2The work of size
The streptomyces species fungus block after mushroom Agaricus blazei Murill fungus block, activation after change.The mushroom Agaricus blazei Murill or streptomyces species
Activation method uses the conventional actication of culture method in this area, including：By the mushroom Agaricus blazei Murill of slant preservation or streptomycete bacterium
Kind is inoculated on PDA plate culture medium, is carried out activation culture, 20 DEG C -30 DEG C of cultivation temperature, incubation time -25 days 3 days, is obtained
The streptomyces species after mushroom Agaricus blazei Murill or activation after activation.Described PDA plate culture medium is trained using this area seed
Conventional culture medium is supported, commercially available prod can be used.Further preferably, described PDA plate culture medium：Potato 200g, grape
Sugared 20g and agar 15g-20g, 1000mL is settled to water.
The volume for the thalline liquid that the inoculum concentration refers to the accession to and the percentage of the ratio between culture volume.
Culture medium used in the present invention uses after being both needed to sterilizing, and the condition of sterilizing uses the normal condition of this area, such as
Can be sterilized 20min-30min at 120 DEG C -125 DEG C.
The composition obtained using medicine fungi fermentation radix tetrastigme is eaten, preferably by the parts by weight radix tetrastigme of 40 parts by weight -55
Fermentation subcritical abstraction thing I and the parts by weight streptomycete fermentation product III of 20 parts by weight -35 form.
The present invention has good hypolipemic function using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, can be used to make
The standby health products or medicine with hypolipemic function.Described health products or medicine can be prepared directly by the composition
Also can be prepared by the composition together with the auxiliary material of this area, product forms can be such as a kind of comprising utilizing with varied
The soft capsule for the composition that medicine fungi fermentation radix tetrastigme obtains is eaten, its content is fermented by the parts by weight radix tetrastigme of 40 parts by weight -55
Subcritical abstraction thing I, the parts by weight streptomycete fermentation product III of 20 parts by weight -35 and the part by weight of vitamin E groups of 10 parts by weight -20
Into.
The bletilla striata (Bletilla striata) also known as Lian Jicao, Gan Gen, Bai Gen, indocalamus orchid, Zhu Lan, purple blue, purple a species of orchid, hundred large bamboo hats with a conical crown and broad brim,
For orchid family bletilla striata platymiscium bletilla striata Bletilla striata (Thunb.) Reichb.f. or narrow leaf bletilla striatas B.ochracca
Schltr. root tuber, underground root tuber is oblate or irregular rhombus, meat, yellow-white.Be mainly distributed on China, Japan and
Upper Myanmar.The bletilla striata has extensive medical value and Ornamental value.Medical value is predominantly used for astringing to arrest bleeding, detumescence and promoting granulation.
Brazilian mushroom (Agaricus blazei Murill), also known as Agricus blazei, originate in the south such as North American southern and Brazil
Beauteously area, the composition of Brazilian mushroom include protein, fat, vitamin, ash content and carbohydrate etc., compared with other bacterium, Brazilian mushroom
The protein and saccharic content of mushroom are relatively abundant, and the compound content of polysaccharide body proteglycan matter is more in fructification.
Streptomycete (Streptomyces) belongs to Gram positive actinomycetes, and substrate mycelium is not broken, and aerial hyphae generally sends out
Educate good, the fibrillae of spores of formation long (sometimes short).Spore can not move, and often have the shape jewelry such as wart, thorn or hair on epitheca.
Raw material used in the present invention can use commercially available prod.
Beneficial effects of the present invention：
1st, when the present invention carries out radix tetrastigme solid state fermentation, directly it is incubated using through the step of liquid MRS culture mediums two containing cholate
The Brazilian mushroom liquid seeds liquid of processing, and the bletilla striata is added into culture medium, can be effectively by the nutritional ingredient in radix tetrastigme
Carry out biodegradable with active ingredient and utilize, effectively the stimulating component in radix tetrastigme is removed and converted, help to carry
The content and activity of active component in high final products.In step (4) and step (5), radix tetrastigme product by solid-state fermentation is carried out
Subcritical Water Extraction, enables that the activity of active ingredient in extract at utmost retains, content is greatly improved；Again will extraction
Residue after the taking culture medium main as streptomycete liquid fermentation, and incubated using through the step of liquid MRS culture mediums two containing NaCl
The streptomycete seed liquor of processing is educated, is recycled by integration, radix tetrastigme resource is fully utilized, is further improved
The content and activity of active component in final products.
2nd, the inventive method raw material sources are natural, quality controllable, environmentally safe, suitable for industrialized production.
3rd, the present invention using eat medicine fungi fermentation radix tetrastigme acquisition composition and common fermentation radix tetrastigme tunning,
Lemna paucicostata is more notable compared to the reduction to T-CHOL and content of triglyceride in serum, illustrates present invention utilization
Composition and its soft gel products that medicine fungi fermentation radix tetrastigme obtains are eaten, T-CHOL in serum and sweet can be significantly reduced
Oily three ester contents, have significant hypolipemic function, can be used to prepare health products or medicine with hypolipemic function.
Embodiment
Present invention is furtherd elucidate with reference to some embodiments, but present disclosure and not only
It is limited to the following examples.
Brazilian mushroom (Agaricus blazei Murill) ACCC50645 strains, are purchased from Chinese agriculture microorganism fungus kind
Preservation administrative center.
Streptomycete (Streptomyces) ACCC40462 strains, are purchased from Chinese agriculture Microbiological Culture Collection administrative center.
Cholate：No. 3 cholate, i.e. sodium taurocholate, cholic acid-NaTDC salt mixture (remote chemical industry in the perseverance industry of Beijing).
Embodiment 1
First, material prepares
PDA plate culture medium：Potato 200g, glucose 20g and agar 15g, 1000ml, natural pH are settled to water,
Sterilize 20min at 121 DEG C.
The streptomyces species of the mushroom Agaricus blazei Murill of slant preservation, slant preservation are inoculated into PDA plate culture medium respectively
On, activation culture is carried out, 22 DEG C of cultivation temperature, incubation time 10 days, respectively obtains the mushroom Agaricus blazei Murill after activation, after activation
Streptomyces species.
The composition of liquid MRS culture mediums is：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, sulphur
Sour 1.0 liters of manganese 0.05g and distilled water.
Fermentation basal medium is made up of the component of following mass percent：Glucose 1%, K2HPO40.1%th, MgSO4
0.05% and the water of surplus.
1L PBSs：Potassium dihydrogen phosphate 0.27g, disodium hydrogen phosphate 1.42g, sodium chloride 8g and potassium chloride 0.2g, add
Appropriate amount of deionized water is sufficiently stirred dissolving, then adds concentrated hydrochloric acid and adjusts pH to 7.4, last constant volume to 1L.
2nd, the preparation of radix tetrastigme fermented product
(1) 2cm is taken2Mushroom Agaricus blazei Murill after size activation is inoculated into 1L liquid MRS culture mediums cultivates 5 in 25 DEG C
My god, nutrient solution collects the first thalline through 6000rpm centrifugations 10min；First thalline is suspended in into cholate weight/mass percentage composition is
10h is incubated in 25 DEG C in the 0.2% liquid MRS culture mediums containing cholate, the second thalline is collected through 6000rpm centrifugations 10min；Will
Second thalline is resuspended in after PBS cleans up in liquid MRS culture mediums, and concussion shakes up to obtain the second thalline
Liquid, the second thalline liquid is inoculated into the liquid MRS containing cholate that cholate weight/mass percentage composition is 0.2% by 5% inoculum concentration and trained
Support and be further cultured in base in 22 DEG C 5 days, obtain Brazilian mushroom liquid seeds liquid；
Wherein, the volume ratio of the first thalline and the liquid MRS culture mediums containing cholate for first thalline that suspends is 1:1,
The volume ratio of second thalline and the liquid MRS culture mediums for second thalline that suspends is 1:1.
(2) 1cm is taken2Streptomyces species after size activation are inoculated into 1L liquid MRS culture mediums cultivates 15h in 25 DEG C,
Nutrient solution collects the 3rd thalline through 6000rpm centrifugations 10min, and it is 5% that the 3rd thalline is suspended in into NaCl weight/mass percentage compositions
8h is incubated in 25 DEG C in liquid MRS culture mediums containing NaCl, the 4th thalline is collected through 6000rpm centrifugations 10min；By the 4th thalline
It is resuspended in after PBS cleans up in liquid MRS culture mediums, concussion shakes up to obtain the 4th thalline liquid, by the 4th
Thalline liquid is inoculated into the liquid MRS culture mediums containing NaCl that NaCl weight/mass percentage compositions are 5% by 3% inoculum concentration in 25 DEG C
15h is cultivated, obtains streptomycete seed liquor；
Wherein, the volume ratio of the 3rd thalline and the liquid MRS culture mediums containing NaCl for the 3rd thalline that suspends is 1:1,
The volume ratio of 4th thalline and the liquid MRS culture mediums for the 4th thalline that suspends is 1:1.
(3) Brazilian mushroom liquid seeds liquid is accessed three by the access amount for accounting for radix tetrastigme solid-state fermentation culture medium volume 15%
In leaf green grass or young crops solid-state fermentation culture medium, cultivated 10 days at 25 DEG C, be subsequently placed in the friendship that magnetic field intensity is 3mT, field frequency is 15Hz
It is further cultured in varying magnetic field environment 20 days, cultured products obtain radix tetrastigme product by solid-state fermentation after vacuum freeze drying, crushing；
The preparation of radix tetrastigme solid-state fermentation culture medium includes：By fresh radix tetrastigme root tuber, bletilla striata root tuber in running water undershoot
Wash clean, dry, cutting is the long sheets of 1cm-2cm or fritter；Again by 8g radix tetrastigmes root tuber and 2g bletilla striatas root tuber fermentation basis
Culture medium is settled to 1L, is well mixed, pH value is naturally, obtain radix tetrastigme solid-state fermentation culture medium.
(4) radix tetrastigme product by solid-state fermentation be crushed into 40 mesh sieves, through Subcritical Water Extraction, the condition of Subcritical Water Extraction
For：Water material mass ratio is 30:1, extracting pressure is 150 DEG C in 15MPa, extraction temperature, extraction time 60min, obtains three leaves
Green grass or young crops fermentation subcritical abstraction thing I, remaining residue obtains extracting residue II after vacuum freeze drying after extraction.
(5) streptomycete seed liquor is accessed into strepto- bacterium solution by the access amount for accounting for streptomycete liquid fermentation medium volume 10%
In body fermentation medium, 60h is cultivated at 25 DEG C, streptomycete fermentation product III is obtained through 6000rpm centrifugations 10min；
The preparation of streptomycete liquid fermentation medium includes：By 0.6g extraction residues II fermentation basal medium constant volume
To 1L, it is well mixed, pH value is naturally, obtain streptomycete liquid fermentation medium.
Fermented the He of subcritical abstraction thing I by 40 parts by weight radix tetrastigmes using the composition of medicine fungi fermentation radix tetrastigme acquisition is eaten
20 parts by weight streptomycete fermentation products III form.
3rd, the preparation of soft gel products：The filler of soft capsule by 40 parts by weight radix tetrastigmes ferment subcritical abstraction thing I,
20 parts by weight streptomycete fermentation products III and 10 part by weight of vitamin E compositions.
Embodiment 2
First, material prepares
PDA plate culture medium：Potato 200g, glucose 20g and agar 20g, 1000ml, natural pH are settled to water,
Sterilize 20min at 121 DEG C.
The streptomyces species of the mushroom Agaricus blazei Murill of slant preservation, slant preservation are inoculated into PDA plate culture medium respectively
On, activation culture is carried out, 20 DEG C of cultivation temperature, incubation time 25 days, respectively obtains the mushroom Agaricus blazei Murill after activation, after activation
Streptomyces species.
Fermentation basal medium is made up of the component of following mass percent：Glucose 3%, K2HPO40.2%th,
MgSO40.1% and the water of surplus.
Liquid MRS culture mediums and PBS are the same as embodiment 1.
2nd, the preparation of radix tetrastigme fermented product
(1) 1cm is taken2Mushroom Agaricus blazei Murill after size activation is inoculated into 1L liquid MRS culture mediums cultivates 8 in 20 DEG C
My god, nutrient solution collects the first thalline through 6000rpm centrifugations 10min；First thalline is suspended in into cholate weight/mass percentage composition is
20h is incubated in 18 DEG C in the 0.05% liquid MRS culture mediums containing cholate, the second thalline is collected through 6000rpm centrifugations 10min；
Second thalline is resuspended in liquid MRS culture mediums after PBS cleans up, concussion shakes up to obtain the second thalline
Liquid, the second thalline liquid is inoculated into the liquid MRS containing cholate that cholate weight/mass percentage composition is 0.05% by 5% inoculum concentration and trained
Support and be further cultured in base in 18 DEG C 10 days, obtain Brazilian mushroom liquid seeds liquid；
Wherein, the volume ratio of the first thalline and the liquid MRS culture mediums containing cholate for first thalline that suspends is 1:2,
The volume ratio of second thalline and the liquid MRS culture mediums for second thalline that suspends is 1:2.
(2) 4cm is taken2Streptomyces species after size activation, which are inoculated into 1L liquid MRS culture mediums, cultivates 3h, training in 30 DEG C
Nutrient solution collects the 3rd thalline through 6000rpm centrifugations 10min, and it is 8% to contain that the 3rd thalline is suspended in into NaCl weight/mass percentage compositions
15h is incubated in 20 DEG C in NaCl liquid MRS culture mediums, the 4th thalline is collected through 6000rpm centrifugations 10min；By the 4th thalline
It is resuspended in after PBS cleans up in liquid MRS culture mediums, concussion shakes up to obtain the 4th thalline liquid, by the 4th
Thalline liquid is inoculated into the liquid MRS culture mediums containing NaCl that NaCl weight/mass percentage compositions are 8% by 3% inoculum concentration in 20 DEG C
30h is cultivated, obtains streptomycete seed liquor；
Wherein, the volume ratio of the 3rd thalline and the liquid MRS culture mediums containing NaCl for the 3rd thalline that suspends is 1:3,
The volume ratio of 4th thalline and the liquid MRS culture mediums for the 4th thalline that suspends is 1:3.
(3) Brazilian mushroom liquid seeds liquid is accessed three by the access amount for accounting for radix tetrastigme solid-state fermentation culture medium volume 25%
In leaf green grass or young crops solid-state fermentation culture medium, cultivated 20 days at 18 DEG C, be subsequently placed in the friendship that magnetic field intensity is 0.1mT, field frequency is 2Hz
It is further cultured in varying magnetic field environment 30 days, cultured products obtain radix tetrastigme product by solid-state fermentation after vacuum freeze drying, crushing；
The preparation of radix tetrastigme solid-state fermentation culture medium includes：By fresh radix tetrastigme root tuber, bletilla striata root tuber in running water undershoot
Wash clean, dry, cutting is the long sheets of 1cm-2cm or fritter；9.5g radix tetrastigmes root tuber and 3.5g bletilla striatas root tuber are fermented again
Basal medium is settled to 1L, is well mixed, pH value is naturally, obtain radix tetrastigme solid-state fermentation culture medium.
(4) radix tetrastigme product by solid-state fermentation be crushed into 40 mesh sieves, through Subcritical Water Extraction, the condition of Subcritical Water Extraction
For：Water material mass ratio is 50:1, extracting pressure is 180 DEG C in 25MPa, extraction temperature, extraction time 10min, obtains three leaves
Green grass or young crops fermentation subcritical abstraction thing I, remaining residue obtains extracting residue II after vacuum freeze drying after extraction.
(5) streptomycete seed liquor is accessed into strepto- bacterium solution by the access amount for accounting for streptomycete liquid fermentation medium volume 15%
In body fermentation medium, 120h is cultivated at 20 DEG C, streptomycete fermentation product III is obtained through 6000rpm centrifugations 10min；
The preparation of streptomycete liquid fermentation medium includes：1g extraction residues II are settled to fermentation basal medium
1L, it is well mixed, pH value is naturally, obtain streptomycete liquid fermentation medium.
Fermented the He of subcritical abstraction thing I by 55 parts by weight radix tetrastigmes using the composition of medicine fungi fermentation radix tetrastigme acquisition is eaten
35 parts by weight streptomycete fermentation products III form.
3rd, the preparation of soft gel products：The filler of soft capsule by 55 parts by weight radix tetrastigmes ferment subcritical abstraction thing I,
35 parts by weight streptomycete fermentation products III and 20 part by weight of vitamin E compositions.
Embodiment 3
First, material prepares
PDA plate culture medium, liquid MRS culture mediums and PBS are the same as embodiment 1.
The streptomyces species of the mushroom Agaricus blazei Murill of slant preservation, slant preservation are inoculated into PDA plate culture medium respectively
On, activation culture is carried out, 30 DEG C of cultivation temperature, incubation time 3 days, respectively obtains the mushroom Agaricus blazei Murill after activation, after activation
Streptomyces species.
Fermentation basal medium is made up of the component of following mass percent：Glucose 2%, K2HPO40.15%th,
MgSO40.08% and the water of surplus.
2nd, the preparation of radix tetrastigme fermented product
(1) 4cm is taken2Mushroom Agaricus blazei Murill after size activation is inoculated into 1L liquid MRS culture mediums cultivates 2 in 30 DEG C
My god, nutrient solution collects the first thalline through 6000rpm centrifugations 10min；First thalline is suspended in into cholate weight/mass percentage composition is
1h is incubated in 30 DEG C in the 0.5% liquid MRS culture mediums containing cholate, the second thalline is collected through 6000rpm centrifugations 10min；Will
Second thalline is resuspended in after PBS cleans up in liquid MRS culture mediums, and concussion shakes up to obtain the second thalline
Liquid, the second thalline liquid is inoculated into the liquid MRS containing cholate that cholate weight/mass percentage composition is 0.5% by 5% inoculum concentration and trained
Support and be further cultured in base in 30 DEG C 1 day, obtain Brazilian mushroom liquid seeds liquid；
Wherein, the volume ratio of the first thalline and the liquid MRS culture mediums containing cholate for first thalline that suspends is 1:3,
The volume ratio of second thalline and the liquid MRS culture mediums for second thalline that suspends is 1:3.
(2) 2cm is taken2Streptomyces species after size activation are inoculated into 1L liquid MRS culture mediums cultivates 30h in 20 DEG C,
Nutrient solution collects the 3rd thalline through 6000rpm centrifugations 10min, and the 3rd thalline is suspended in into NaCl weight/mass percentage compositions for 0.5%
The liquid MRS culture mediums containing NaCl in 30 DEG C be incubated 0.5h, through 6000rpm centrifugation 10min collect the 4th thalline；By the 4th
Thalline is resuspended in after PBS cleans up in liquid MRS culture mediums, and concussion shakes up to obtain the 4th thalline liquid, will
4th thalline liquid is inoculated into by 3% inoculum concentration in the liquid MRS culture mediums containing NaCl that NaCl weight/mass percentage compositions are 0.5%
3h is cultivated in 30 DEG C, obtains streptomycete seed liquor；
Wherein, the volume ratio of the 3rd thalline and the liquid MRS culture mediums containing NaCl for the 3rd thalline that suspends is 1:2,
The volume ratio of 4th thalline and the liquid MRS culture mediums for the 4th thalline that suspends is 1:2.
(3) Brazilian mushroom liquid seeds liquid is accessed three by the access amount for accounting for radix tetrastigme solid-state fermentation culture medium volume 5%
In leaf green grass or young crops solid-state fermentation culture medium, cultivated 2 days at 30 DEG C, be subsequently placed in the alternation that magnetic field intensity is 5mT, field frequency is 30Hz
It is further cultured in magnetic field environment 5 days, cultured products obtain radix tetrastigme product by solid-state fermentation after vacuum freeze drying, crushing；
The preparation of radix tetrastigme solid-state fermentation culture medium includes：By fresh radix tetrastigme root tuber, bletilla striata root tuber in running water undershoot
Wash clean, dry, cutting is the long sheets of 1cm-2cm or fritter；6.5g radix tetrastigmes root tuber and 0.5g bletilla striatas root tuber are fermented again
Basal medium is settled to 1L, is well mixed, pH value is naturally, obtain radix tetrastigme solid-state fermentation culture medium.
(4) radix tetrastigme product by solid-state fermentation be crushed into 40 mesh sieves, through Subcritical Water Extraction, the condition of Subcritical Water Extraction
For：Water material mass ratio is 10:1, extracting pressure is 100 DEG C in 4MPa, extraction temperature, extraction time 90min, obtains radix tetrastigme
Ferment subcritical abstraction thing I, and remaining residue obtains extracting residue II after vacuum freeze drying after extraction.
(5) streptomycete seed liquor is accessed into strepto- bacterium solution by the access amount for accounting for streptomycete liquid fermentation medium volume 3%
In body fermentation medium, 10h is cultivated at 30 DEG C, streptomycete fermentation product III is obtained through 6000rpm centrifugations 10min；
The preparation of streptomycete liquid fermentation medium includes：By 0.2g extraction residues II fermentation basal medium constant volume
To 1L, it is well mixed, pH value is naturally, obtain streptomycete liquid fermentation medium.
Fermented the He of subcritical abstraction thing I by 50 parts by weight radix tetrastigmes using the composition of medicine fungi fermentation radix tetrastigme acquisition is eaten
25 parts by weight streptomycete fermentation products III form.
3rd, the preparation of soft gel products：The filler of soft capsule by 50 parts by weight radix tetrastigmes ferment subcritical abstraction thing I,
25 parts by weight streptomycete fermentation products III and 15 part by weight of vitamin E compositions.
Comparative example 1
First, material prepares
Mushroom Agaricus blazei Murill, liquid MRS culture mediums and fermentation basal medium after PDA plate culture medium, activation is same
Embodiment 1.
2nd, the preparation of radix tetrastigme fermented product
(1) 2cm is taken2Mushroom Agaricus blazei Murill after size activation is inoculated into 1L liquid MRS culture mediums cultivates 5 in 25 DEG C
My god, obtain Brazilian mushroom liquid seeds liquid.
(2) Brazilian mushroom liquid seeds liquid is accessed into radix tetrastigme by the access amount for accounting for radix tetrastigme fermentation medium volume 15%
In fermentation medium, cultivated 10 days at 25 DEG C, cultured products obtain radix tetrastigme tunning after vacuum freeze drying, crushing；
The preparation of radix tetrastigme fermentation medium includes：Fresh radix tetrastigme root tuber is rinsed well under running water, dries, cuts
It is divided into the long sheets of 1cm-2cm or fritter；8g radix tetrastigmes root tuber is settled to 1L with fermentation basal medium again, is well mixed, pH
Value is naturally, obtain radix tetrastigme fermentation medium.
3rd, the preparation of soft gel products：The filler of soft capsule is by 40 parts by weight radix tetrastigme tunnings and 10 parts by weight
Vitamin E forms.
Comparative example 2
Fresh radix tetrastigme root tuber is rinsed well under running water, dried, cutting is the long sheets of 1cm-2cm or fritter；Press
Feed liquid weight is than 1:10 add water, extract 10h under fluidized state, filter to obtain first-time filtrate and a filter residue, a filter residue is pressed
Feed liquid weight is than 1:5 add water, and 8h is extracted under fluidized state, filter to obtain secondary filtrate, merge first-time filtrate and secondary filtrate, warp
Freeze-drying, obtain Lemna paucicostata after crushing.
The preparation of soft gel products：The filler of soft capsule is by 40 parts by weight Lemna paucicostatas and 10 part by weight of vitamin
E is formed.
The measure of active component
(1) measurement of the polysaccharide content
Appropriate amount of sample dry powder is weighed, 40 mesh are crossed after crushing, are placed in beaker, by solid-liquid ratio 1:10 (mass ratioes) add distillation
Water, 90 DEG C of extraction 3h, extraction twice, are filtered under diminished pressure and collect filtrate, be concentrated under reduced pressure at 50 DEG C, it is anhydrous that concentrate adds 4 times of volumes
Ethanol, alcohol precipitation overnight, after 3000r/min, 20min centrifugation, precipitation is Thick many candies.Precipitation is settled to 50mL with distilled water,
Polysaccharide in fermentation liquid content is determined with phend-sulphuric acid, replication is averaged for 3 times, i.e. polyoses content.
(2) measure of flavones content
Accurate weigh is dried to the rutin standard items 10.0mg of constant weight, with mass percentage concentration 70% under the conditions of 120 DEG C
Ethanol water dissolves and is settled to 100mL, is made into 0.1mg/mL standard liquids.Take respectively standard liquid 0.0,1.0,2.0,
3.0th, 4.0,5.0mL is in 10mL volumetric flasks, adds the ethanol water of mass percentage concentration 70% respectively to after 5.0mL, adds
The NaNO of 0.3mL mass percentage concentrations 5%2The aqueous solution, shake up and place 6min；Again plus 0.4mL mass percentage concentrations 10% Al
(NO3)3The aqueous solution, shake up and place 6min；Again plus the 4.0mL mass percentage concentration 4%NaOH aqueous solution, mass percentage concentration 60%
Ethanol water is settled to scale, shakes up and places 15min, is cooled to the absorbance of the bioassay standard product at 510nm after room temperature, with
Standard solution mass concentration C (mg/mL) is abscissa, and absorbance A is that ordinate draws standard curve, obtains regression equation.
Appropriate amount of sample dry powder is weighed, 40 mesh are crossed after crushing, first by solid-liquid ratio 1:30 (mass ratioes) add mass percentage concentration
70% ethanol water, 2h is extracted at 70 DEG C, 50ml is settled to after extract solution centrifugal filtration, therefrom take 1ml samples to carry out
Assay.
1 each embodiment of table and comparative example main component testing result
Note：Compared with comparative example 1, comparative example 2, Δ Δ：P<0.01.
Shown by the data of table 1, the present invention is significantly carried simultaneously using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme
Polysaccharide, the content of flavones isoreactivity composition in high product.Compared with the conventional culture methods of comparative example 1, using the inventive method
Polyoses content improves 68.2%-71.1% in the radix tetrastigme composition of acquisition, and flavones content improves 80.6%-92.2%.
Aid in lipid-lowering test
(1) experimental animal is grouped
Mouse 24-28g, male and female half and half, adaptability are grouped at random after growing 1 week according to TC is horizontal, every group 10.Pass through survey
Determine the horizontal effect for judging reducing blood lipid of serum total cholesterol (TC), triglycerides (TG).
(2) medication and dosage
High lipid food be Fukang bio tech ltd of Beijing China KK mouse material, crude fat weight/mass percentage composition
16.15%, given the test agent (the i.e. composition, contrast in embodiment 1-3 of various dose are given while high lipid food is given
The Lemna paucicostata in radix tetrastigme tunning and comparative example 1 in example 1).Experiment sets 5 processing, i.e., high, medium and low 3 agent
Amount group (8.0ml/kg, 16.0ml/kg, 24.0ml/kg), Basal control group and high fat control group, high fat control group and basis are right
According to group with distilled water gavage, and high lipid food and basal feed (being purchased from Guangdong Province medical animal field) are given respectively, with dosage
Group is synchronous to be carried out.Each daily 1 soft capsule sample of gavage of dosage group, continuous feeding 30 days, feeds while given the test agent is given
High lipid food, ad lib and drinking-water.
(3) content and method are determined
This experiment is preventive administration, the given the test agent of various dose is given while high lipid food is given, in experiment
Terminate fasting 16 hours, kit (COD-PAP methods) and triglyceride determination kit (GPO-PAP are determined according to T-CHOL
Method) in described in specification, it is horizontal to survey serum total cholesterol (TC), triglycerides (TG) respectively.Testing result is shown in Table 2.
Influence (X ± S, n=10) of the soft capsule sample of table 2 to mouse TC and TG
△ represents P ＜ 0.05, significant difference compared with high fat control group；* the P compared with the matched doses group of comparative example 1 is represented
＜ 0.05, significant difference.
The data of table 2 are shown, common using the composition and comparative example 1 for eating the acquisition of medicine fungi fermentation radix tetrastigme using the present invention
Radix tetrastigme tunning, the Lemna paucicostata of comparative example 2 of fermentation are compared to the T-CHOL and content of triglyceride in serum
Reduction it is more notable, the present invention utilizes the composition drop serum total cholesterol (TC), sweet for eating the acquisition of medicine fungi fermentation radix tetrastigme
The effect of oily three esters (TG) will be substantially better than the radix tetrastigme tunning of the common fermentation of comparative example 1, the extraction of the radix tetrastigme of comparative example 2
Thing, illustrate that the present invention using the composition and its soft gel products of eating the acquisition of medicine fungi fermentation radix tetrastigme, can significantly reduce blood
T-CHOL and content of triglyceride in clear, have significant hypolipemic function.
The change of each parameter has no effect on of the invention utilize and eats medicine in the range of inventive formulation and preparation method limit
Blood fat reducing function and the preparation of composition and its soft gel products that fungi fermentation radix tetrastigme obtains, therefore inventive formulation and system
The combination of arbitrary parameter can obtain utilizing the composition and its soft capsule production for eating the acquisition of medicine fungi fermentation radix tetrastigme in Preparation Method
Product.It will not be repeated here.
Claims (10)
1. a kind of preparation method using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, it is characterised in that including step：
(1) Brazilian mushroom (Agaricus blazei Murill) strain after activation is taken to be inoculated into liquid MRS culture mediums and train
Support -8 days 2 days, the first thalline is collected by centrifugation；First thalline is suspended in the liquid MRS culture mediums containing cholate in 18 DEG C -30 DEG C
1h-20h is incubated, the second thalline is collected by centrifugation；Second thalline is resuspended in liquid MRS after PBS cleans up
In culture medium, concussion is shaken up to obtain the second thalline liquid, and the second thalline liquid is inoculated into the liquid MRS culture mediums containing cholate in 18
DEG C -30 DEG C are further cultured for -10 days 1 day, obtain Brazilian mushroom liquid seeds liquid；
(2) take streptomycete (Streptomyces) strain after activation to be inoculated into liquid MRS culture mediums and cultivate 3h-30h, centrifuge
The 3rd thalline is collected, the 3rd thalline is suspended in the liquid MRS culture mediums containing NaCl in 20 DEG C of -30 DEG C of incubation 0.5h-15h,
The 4th thalline is collected by centrifugation；4th thalline is resuspended in after PBS cleans up in liquid MRS culture mediums, shaken
Swing and shake up to obtain the 4th thalline liquid, the 4th thalline liquid is inoculated into the liquid MRS culture mediums containing NaCl in 20 DEG C of -30 DEG C of cultures
3h-30h, obtain streptomycete seed liquor；
(3) by the Brazilian mushroom liquid seeds liquid access radix tetrastigme solid-state fermentation culture medium of step (1), trained at 18 DEG C -30 DEG C
Support -20 days 2 days, be subsequently placed in alternating magnetic field environment and be further cultured for -30 days 5 days, cultured products are after vacuum freeze drying, crushing
Obtain radix tetrastigme product by solid-state fermentation；Contain radix tetrastigme and the bletilla striata in the radix tetrastigme solid-state fermentation culture medium；
(4) the radix tetrastigme product by solid-state fermentation of step (3) is obtained into radix tetrastigme fermentation subcritical abstraction thing through Subcritical Water Extraction
I, remaining residue obtains extracting residue II after vacuum freeze drying after extraction；
(5) by the streptomycete seed liquor access streptomycete liquid fermentation medium of step (2), 10h- are cultivated at 20 DEG C -30 DEG C
120h, centrifugation, streptomycete fermentation product III is obtained, contains extraction residue II in the streptomycete liquid fermentation medium；
It is described to be sent out using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme for radix tetrastigme fermentation subcritical abstraction thing I and streptomycete
Ferment product III.
2. preparation method according to claim 1, it is characterised in that in step (3), the radix tetrastigme solid state fermentation culture
Contain 6.5g-9.5g radix tetrastigmes root tuber and 0.5g-3.5g bletilla striata root tubers in the every 1L of base.
3. preparation method according to claim 1, it is characterised in that in step (5), the streptomycete liquid fermentation and culture
Contain 0.2g-1g extraction residues II in the every 1L of base.
4. preparation method according to claim 1, it is characterised in that in step (1), the liquid MRS trainings containing cholate
The weight/mass percentage composition for supporting cholate in base is 0.05%-0.5%；
In step (2), the weight/mass percentage composition of the liquid MRS NaCl in medium containing NaCl is 0.5%-8%.
5. preparation method according to claim 1, it is characterised in that in step (1), first thalline suspends with being used for
The volume ratio of the liquid MRS culture mediums containing cholate of first thalline is 1:1-3, the second thalline and the liquid for second thalline that suspends
The volume ratio of body MRS culture mediums is 1:1-3, the inoculum concentration of the second thalline liquid is 5%；
In step (2), the volume ratio of the 3rd thalline and the liquid MRS culture mediums containing NaCl for the 3rd thalline that suspends is
1:The volume ratio of 1-3, the 4th thalline and the liquid MRS culture mediums for the 4th thalline that suspends is 1:1-3, the 4th thalline liquid connect
Kind amount is 3%.
6. preparation method according to claim 1, it is characterised in that in step (3), magnetic field in the alternating magnetic field environment
Intensity is 0.1mT-5mT, field frequency 2Hz-30Hz；
In step (4), the condition of the Subcritical Water Extraction is：Water material mass ratio is 10-50:1, extracting pressure is in 4MPa-
25MPa, extraction temperature are 100 DEG C -180 DEG C, extraction time 10min-90min.
7. preparation method according to claim 1, it is characterised in that in step (3), Brazilian mushroom liquid seeds liquid connects
Enter the 5%-25% that amount is radix tetrastigme solid-state fermentation culture medium volume；
In step (5), the access amount of the streptomycete seed liquor is the 3%-15% of streptomycete liquid fermentation medium volume.
8. a kind of utilize the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, it is characterised in that using any one of claim 1-7
Described preparation method is prepared.
9. according to claim 8 utilize the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, it is characterised in that described group
Compound is produced by the parts by weight radix tetrastigme of 40 parts by weight -55 fermentation subcritical abstraction thing I and the parts by weight streptomycete fermentation of 20 parts by weight -35
Thing III forms.
10. a kind of application using the composition for eating the acquisition of medicine fungi fermentation radix tetrastigme, it is characterised in that according to claim 8
Or the composition of medicine fungi fermentation radix tetrastigme acquisition answering in the health products or medicine for preparing reducing blood lipid is eaten in the utilization described in 9
With.
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| 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 |
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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 |
|---|
| Toxicological Effects of the Extract of Tetrastigma hemsleyanum Diels et. Gilg on Hepatocellular Carcinoma Cell Line HepG_2 and Primary Rat Hepatocytes in vitro;DING Gang-qiang，etal;《Zhejiang Journal of Preventive Medicine》;20051231;1-5 * |
| 中国特有植物三叶青化学成分的研究;刘东，等;《中国中药杂志》;19991231;第611-612页 * |
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