CN103911294B - One seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method - Google Patents
One seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method Download PDFInfo
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Abstract
The invention provides a seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method, belong to Chinese medicine microorganism fermentation field. The method is by the lucidum strain being stored in PDA solid medium, in the liquid fermentation medium that to be seeded to Chinese medicine ginseng, water extraction of astragalus membranaceus be matrix, be that 22～30 DEG C, rotating speed are to cultivate 10d～20d under 100～180r/min condition in temperature, obtain joining stilbene sesame medicinal fungal substance. The liquid fermentation medium that the present invention uses is ginseng, the composite Chinese traditional medicine water extract of the Radix Astragali, adds without other chemical compositions, safe and harmless; The combination that people is participated in to the Radix Astragali will make up the deficiency of the property of medicine, drug effect separately, expand the scope of indication, and under the fermentation of glossy ganoderma, in the medicinal fungal substance obtaining, there is the active component of ginseng, the Radix Astragali, glossy ganoderma simultaneously, realize effective compatibility of 3 taste traditional Chinese medicines, pharmacological property balance, the biologically active of lifting crude drug.
Description
Technical field
The invention belongs to Chinese medicine microorganism fermentation field, be specifically related to a seed ginseng stilbene sesame medicinal fungal substance liquid fermentationMethod.
Background technology
Ginseng, the Radix Astragali are all Qi-tonifying drugs conventional in clinical treatment, and still, in the property of medicine, drug effect, the two is depositedIn difference. Ginseng sweet and slightly bitter taste and property are flat, and the tonifying Qi yin-nourishing of holding concurrently is kept and do not walked. Radix Astragali taste is sweet warm in nature, tonifying QiThe double sun of holding up, walks and does not keep. By both pairings, ginseng stilbene one is kept one and is walked, and negative and positive are taken into account, logical benefit nothing out and outRush down, can be used for the disease of all deficiency of vital energy deficiencies, there is powerful tonifying Qi and help unit's effect. Medicinal fungi glossy ganoderma is toolTraditional rare medicinal herbs of have strengthening by means of tonics, strengthening the body resistance to consolidate the constitution, modern pharmacology studies confirm that, glossy ganoderma is for improvementMemory, promotes sleep, strengthens body immunity, auxiliary tumor chemoradiotherapy, the equal tool in the aspects such as liver protectingThere is significant curative effect.
The two-way fermentation technique of Chinese medicine is medicinal fungi is seeded in Chinese medicine matrix, fermenting and producing medicinal fungiTime, the chemical composition that the enzyme of generation can catalysis Chinese medicine transforms, and improves or reduce chemical composition of Chinese materia medicaContent or produce noval chemical compound, biologically active, the toxicity of the medicinal fungal substance that obtains of fermentation are changed,Or possesses new activity. Because liquid fermentation is short compared with solid fermentation required time, output large, product qualityStable, production process is easy to monitoring, batch production industrialization level is high, therefore two-way fermentation technique is applied toLiquid fermentation has larger economy and social benefit in producing. The literary composition of the liquid bidirectional fermentation of having reported at presentOffer lessly, wherein Chinese patent 200910218080.7 discloses a kind of glossy ganoderma fermentation red ginseng decocting liquid of utilizingThe anti-fatigue active of optimum process condition and its tunning. Not yet have at present any document open taking glossy ganoderma asFermented bacterium, fermentation is with the two-way liquid fermentation work of the Compositional type fermentation substrate of 2 kinds of Chinese traditional medicine composition of ginseng-Radix AstragaliThe application of skill and the anti-oxidant aspect of medicinal fungal substance thereof.
Summary of the invention
The object of this invention is to provide a seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method, the method time is short,And the ginseng stilbene sesame mycoplasma obtaining has higher antioxidation activity.
The invention provides a seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method, the method comprises:
To be stored in lucidum strain in PDA solid medium by 3mm × 3mm, be seeded to Chinese medicine ginseng,Water extraction of astragalus membranaceus is in the liquid fermentation medium of matrix, and 2 every bottle is that 22～30 DEG C, rotating speed are in temperatureUnder 100～180r/min condition, cultivate 10d～20d, obtain joining stilbene sesame medicinal fungal substance;
Described liquid fermentation medium is prepared as follows: Chinese medicine ginseng, the Radix Astragali pulverized, crossed 10 mesh sieves,Take respectively ginseng 20～60g, the Radix Astragali 6～90g, both mix, 300～500ml distilled water heating and refluxing extraction2～3 times, each 0.5h, merges water extraction liquid and is settled to 1L, and pH nature, is sub-packed in 250ml triangular flask, everyBottled liquid measure 50ml, sealing, 115～121 DEG C of sterilizing 20～30min.
Preferably, described lucidum strain comprises red sesame and cold sesame.
Preferably, when described lucidum strain is red sesame, in liquid fermentation medium, get ginseng 30g, HuangStilbene 10g.
Preferably, when described lucidum strain is red sesame, fermentation temperature is 24～26 DEG C, and rotating speed is120～160r/min。
Preferably, when described lucidum strain is red sesame, fermentation time is 15d～17d.
Preferably, when described lucidum strain is cold sesame, in liquid fermentation medium, get ginseng 30g, HuangStilbene 15g.
Preferably, when described lucidum strain is cold sesame, fermentation temperature is 26～28 DEG C, and rotating speed is120～160r/min。
Preferably, when described lucidum strain is cold sesame, fermentation time is 15d.
Beneficial effect of the present invention
The invention provides a seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method, the liquid fermentation training that the present invention usesFoster base is ginseng, the composite Chinese traditional medicine water extract of the Radix Astragali, adds without other chemical compositions, safe and harmless; By ginsengTo make up the deficiency of the property of medicine, drug effect separately with the combination of the Radix Astragali, expand the scope of indication, and glossy ganodermaFermentation under, in the medicinal fungal substance of acquisition, there is the active component of ginseng, the Radix Astragali, glossy ganoderma simultaneously, realize 3 tastesEffective compatibility of traditional Chinese medicine, pharmacological property balance, the biologically active of lifting crude drug. By gained zymotic fluidAnd mycelium separates, the ultrasonic extraction of methanol solution after zymotic fluid freeze-drying; Mycelium crushed after being dried, 70%The ultrasonic extraction of ethanol, respectively with fermentation broth extract and hypha extract remove ultra-oxygen anion free radical andHydroxyl radical free radical, when result shows 10mg/ml, red sesame zymotic fluid is clear to superoxide anion and hydroxyl radical free radicalExcept rate is for being not less than 70% and 50%; Red camphorata mycelium is not to the clearance rate of superoxide anion and hydroxyl radical free radical allLower than 30%; Cold sesame zymotic fluid the clearance rate of superoxide anion and hydroxyl radical free radical is not less than and 45% and50%; Cold camphorata mycelium is not less than 50% and 10% to the clearance rate of superoxide anion and hydroxyl radical free radical, visibleThe ginseng stilbene sesame medicinal fungal substance that the inventive method makes has stronger antioxidation activity.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is done to further detailed description.
Comparative example 1
Chinese medicine ginseng, the Radix Astragali are pulverized, are crossed 10 mesh sieves, take respectively ginseng and Radix Astragali 20g, 30g, 40g,50g, 60g, add respectively 500ml distilled water heating and refluxing extraction 2 times, and each 0.5h, merges water extraction liquidBe settled to 1L, pH nature, is sub-packed in 250ml triangular flask every bottled liquid measure 50ml. Sealing, 121DEG C sterilizing 20min, cooling for subsequent use. To be stored in the red sesame in PDA solid medium, cold sesame bacterial classification by 3mm× 3mm, is seeded in the liquid fermentation medium that Chinese medicine ginseng or water extraction of astragalus membranaceus are matrix, and 2 every bottle,Under 26 DEG C, 150r/min condition, cultivate 12d, zymotic fluid after filtration, and rinses three times repeatedly, filters, dry,Obtain mycelium. The mycelia scale of construction data that comparative example 1 obtains are as shown in table 1:
Table 1
The mycelia scale of construction of table 1 for fermenting red sesame according to concentration fermentation substrate in difference, trembling with fear after sesame 12d, fromTable 1 can be found out: water extraction of astragalus membranaceus is discontented with the demand of pure gold sesame, cold sesame growth, and the mycelia scale of construction is less;Ginseng aqueous extract is suitable for growth demand, especially when concentration in ginseng aqueous extract is 30g～60g/LTime, the accumulation of bacterial classification mycelial biomass is higher. So by 30～60g/L ginseng aqueous extract (especially 30g/LGinseng aqueous extract) as the main matrix in composite Chinese medicine matrix, adding wherein water extraction of astragalus membranaceus, compositionComposite herb fermenting matrix.
Embodiment 1
Chinese medicine ginseng, the Radix Astragali are pulverized, are crossed 10 mesh sieves, take 5 parts of 30g ginsengs, then take respectively Radix Astragali 6g,10g, 15g, 30g, 60g, 90g, mix with ginseng respectively, adds 500ml distilled water heating and refluxing extraction 2Inferior, each 0.5h, merges water extraction liquid and is settled to 1L, and pH nature, is sub-packed in 250ml triangular flask, often bottledLiquid measure 50ml, sealing, 121 DEG C of sterilizing 20min, cooling for subsequent use. To be stored in PDA solid mediumRed sesame, cold sesame bacterial classification are pressed 3mm × 3mm, are seeded to Chinese medicine ginseng, the full medicinal material water extraction of Radix Astragali liquid is matrixIn liquid fermentation medium, 2 every bottle, under 26 DEG C, 150r/min condition, cultivate 12d, zymotic fluid after filtration,And repeatedly rinse three times, filtering, dry, obtain joining stilbene sesame property of medicine mycelium. The bacterium that embodiment 1 obtainsFilament amount data are as shown in table 2:
Table 2
Embodiment 2
Chinese medicine ginseng, the Radix Astragali are pulverized, crossed 10 mesh sieves, take ginseng 30g, Radix Astragali 10g is red sesame fermentation baseMatter; Take ginseng 30g, Radix Astragali 15g, for cold sesame Zymocyte, adds respectively 500ml distilled water to add hot reflux and carriesGet 2 times, each 0.5h, merges water extraction liquid and is settled to 1L, and pH nature, is sub-packed in 250ml triangular flask, everyBottled liquid measure 50ml. Sealing, 121 DEG C of sterilizing 20min, cooling for subsequent use. To be stored in PDA solid mediumIn red sesame, cold sesame bacterial classification press 3mm × 3mm, be seeded to Chinese medicine ginseng, the full medicinal material water extraction of Radix Astragali liquid is baseIn the liquid fermentation medium of matter, 2 every bottle, adjusting temperature is 22 DEG C, 24 DEG C, 26 DEG C, 28 DEG C, 30 DEG C,Under 150r/min condition, cultivate 12d, zymotic fluid after filtration, and rinses three times repeatedly, is filtering, dry,Obtain joining stilbene sesame property of medicine mycelium. The mycelia scale of construction data that embodiment 2 obtains are as shown in table 3:
Table 3
Embodiment 3
Chinese medicine ginseng, the Radix Astragali are pulverized, crossed 10 mesh sieves, take ginseng 30g, Radix Astragali 10g is red sesame fermentation baseMatter; Take ginseng 30g, Radix Astragali 15g, for cold sesame Zymocyte, adds respectively 300ml distilled water to add hot reflux and carriesGet 3 times, each 0.5h, merges water extraction liquid and is settled to 1L, and pH nature, is sub-packed in 250ml triangular flask, everyBottled liquid measure 50ml. Sealing, 121 DEG C of sterilizing 20min, cooling for subsequent use. To be stored in PDA solid mediumIn red sesame, cold sesame bacterial classification press 3mm × 3mm, be seeded to Chinese medicine ginseng, the full medicinal material water extraction of Radix Astragali liquid is baseIn the liquid fermentation medium of matter, 2 every bottle, adjust rotating speed to 100,120,140,160,180r/min,Under 26 DEG C of conditions, cultivate 12d, zymotic fluid after filtration, and rinses three times repeatedly, is filtering, dry, obtainsGinseng stilbene sesame property of medicine mycelium. The mycelia scale of construction data that embodiment 3 obtains are as shown in table 4:
Table 4
Embodiment 4
Chinese medicine ginseng, the Radix Astragali are pulverized, crossed 10 mesh sieves, take ginseng 30g, Radix Astragali 10g is red sesame fermentation baseMatter; Take ginseng 30g, Radix Astragali 15g, for cold sesame Zymocyte, adds respectively 500ml distilled water to add hot reflux and carriesGet 2 times, each 0.5h, merges water extraction liquid and is settled to 1L, and pH nature, is sub-packed in 250ml triangular flask, everyBottled liquid measure 50ml. Sealing, 121 DEG C of sterilizing 20min, cooling for subsequent use. To be stored in PDA solid mediumIn red sesame, cold sesame bacterial classification press 3mm × 3mm, be seeded to Chinese medicine ginseng, the full medicinal material water extraction of Radix Astragali liquid is baseIn the liquid fermentation medium of matter, 2 every bottle, 26 DEG C, 150r/min condition fermentation, get cultivate 11d, 13d,15d, 17d, 19d fermented and cultured bottle, rinse separation and fermentation liquid and mycelium after filtration, repeatedly. Zymotic fluid freezesThe ultrasonic extraction of methanol solution after dry; Mycelium crushed after being dried, the ultrasonic extraction of 70% ethanol, respectively with zymotic fluidExtract and hypha extract are removed ultra-oxygen anion free radical and hydroxyl radical free radical. Data are as shown in table 5:
Table 5
The explanation of above embodiment is just for helping to understand method of the present invention and core concept thereof. Should refer toGo out, for those skilled in the art, under the premise without departing from the principles of the invention, alsoCan carry out some improvement and modification to the present invention, these improvement and modify and also fall into the claims in the present inventionIn protection domain.
To the above-mentioned explanation of the disclosed embodiments, make professional and technical personnel in the field can realize or use thisInvention. To be apparent for those skilled in the art to the multiple amendment of these embodiment,General Principle as defined herein can be in the situation that not departing from the spirit or scope of the present invention, at otherIn embodiment, realize. Therefore, the present invention will can not be restricted to these embodiment shown in this article, but wantMeet the widest scope consistent with principle disclosed herein and features of novelty.
Claims (8)
1. a seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method, is characterized in that, the method comprises:
To be stored in lucidum strain in PDA solid medium by 3mm × 3mm, in the liquid fermentation medium that to be seeded to Chinese medicine ginseng, water extraction of astragalus membranaceus be matrix, 2 every bottle is that 22～30 DEG C, rotating speed are to cultivate 10d～20d under 100～180r/min condition in temperature, obtains joining stilbene sesame medicinal fungal substance;
Described liquid fermentation medium is prepared as follows: Chinese medicine ginseng, the Radix Astragali are pulverized, cross 10 mesh sieves, take respectively ginseng 20～60g, the Radix Astragali 6～90g, both mix, 300～500ml distilled water heating and refluxing extraction 2～3 times, each 0.5h, merges water extraction liquid and is settled to 1L, pH nature, be sub-packed in 250ml triangular flask, every bottled liquid measure 50ml, sealing, 115～121 DEG C of sterilizing 20～30min.
2. according to the seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method shown in claim 1, it is characterized in that, described lucidum strain comprises red sesame and cold sesame.
3. according to the seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method shown in claim 1, it is characterized in that, when described lucidum strain is red sesame, in liquid fermentation medium, get ginseng 30g, Radix Astragali 10g.
4. according to the seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method shown in claim 1, it is characterized in that, when described lucidum strain is red sesame, fermentation temperature is 24～26 DEG C, and rotating speed is 120～160r/min.
5. according to the seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method shown in claim 1, it is characterized in that, when described lucidum strain is red sesame, fermentation time is 15d～17d.
6. according to the seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method shown in claim 1, it is characterized in that, when described lucidum strain is cold sesame, in liquid fermentation medium, get ginseng 30g, Radix Astragali 15g.
7. according to the seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method shown in claim 1, it is characterized in that, when described lucidum strain is cold sesame, fermentation temperature is 26～28 DEG C, and rotating speed is 120～160r/min.
8. according to the seed ginseng stilbene sesame medicinal fungal substance liquid fermentation method shown in claim 1, it is characterized in that, when described lucidum strain is cold sesame, fermentation time is 15d.
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