CN109825533A - A kind of fermentation medium and its fermentation culture method being used to prepare high activity Phellinus tunning - Google Patents
A kind of fermentation medium and its fermentation culture method being used to prepare high activity Phellinus tunning Download PDFInfo
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Abstract
The invention discloses a kind of fermentation mediums and its fermentation culture method for being used to prepare high activity Phellinus tunning; wherein fermentation medium is by weight; be grouped as by following group: big bean sprout starches 20-30 parts; 5-10 parts of Semen Tritici aestivi fiber element, 5-10 parts of radix tetrastigme zymolyte, ramulus mori and 2-3 parts of blue or green money willow extract; 0.05-0.1 parts of calcium chloride, 0.3-0.5 parts of lactose; 1-3 parts of glucose, 0.02-0.05 parts of salicylic acid, 80-100 parts of water.Fermentation medium of the invention helps to improve the content and activity of active constituent in final products Phellinus tunning.The high activity Phellinus tunning of preparation has significant protective effect to the subacute oxidative damage of D- galactolipin inducing mouse.
Description
Technical field
The present invention relates to a kind of fermentation mediums and its fermentation culture method for being used to prepare high activity Phellinus tunning.
Background technique
Phellinus (Inonotus sanghuang) parasitizes on mulberry tree, and fructification cap majority is storied together, horseshoe
Shape or irregular shape, children make lemon yellow to golden yellow, khaki or yellowish-brown when old.Phellinus is a kind of perennial large size of preciousness
Medicinal wood-rotting fungi, is usually grown on Mulberry plant.Phellinus is known as " forest gold " by modern.Phellinus is current international public
Efficient highest a kind of medicinal fungi in the biological anticancer preparation recognized.China is Wild Medicinal fungi big export country, but at present
To the research of Phellinus bacterium also in the elementary step, technology is not yet mature.Since China's limited Phellinus wild resource growth period is long,
Functional component stability is poor, and is excessively acquired, fewer and fewer, it is difficult to as stable industrial products source, far can not expire
The demand in sufficient market.Prepare biological products using biofermentation technique, be overcome using wild resource biomass be limited it is best
Approach and the basis for realizing desirable metabolites development and utilization.It is compared with the cultivation of traditional fructification, mulberry is prepared by liquid fermentation
Yellow secondary metabolite has many advantages, such as that with short production cycle, metabolite is more, quality is easily controllable, large-scale production.Use liquid
The fermentation medium of fermentation preparation Phellinus secondary metabolite is of great significance, and can directly affect the production of Phellinus secondary metabolism
The activity of object.
Summary of the invention
In view of this, it is an object of the invention to propose a kind of fermented and cultured for being used to prepare high activity Phellinus tunning
Base and its fermentation culture method can effectively improve the activity of Phellinus tunning.
Used technical solution are as follows:
A kind of fermentation medium being used to prepare high activity Phellinus tunning is grouped as by following group by weight:
Big bean sprout starches 20-30 parts, and 5-10 parts of Semen Tritici aestivi fiber element, 5-10 parts of radix tetrastigme zymolyte, ramulus mori and 2-3 parts of blue or green money willow extract,
0.05-0.1 parts of calcium chloride, 0.3-0.5 parts of lactose, 1-3 parts of glucose, 0.02-0.05 parts of salicylic acid, 80-100 parts of water.
Further, big bean sprout slurry is prepared via a method which: select the complete soybean of grain shape, be put into it is complete from
Vernalization is sprayed in dynamic bean sprout growing machine, when soybean bud head is revealed to 4-5mm, stops germination；By big bean sprout: water is mixed by weight 1: 1,
High speed mashing obtains big bean sprout slurry.
Further, the ramulus mori and blue or green money willow extract are prepared via a method which: respectively by ramulus mori and green money
Willow leaf crushes, and mixes by weight 1:1 ratio, the ethanol solution of the 70vol% of 20-30 times of weight is added, in 40-50 DEG C shake
Concussion is extracted 10 hours in bed, and supernatant is obtained after filtering and after vacuum freeze drying, obtains ramulus mori after supernatant is concentrated under reduced pressure
With blue or green money willow extract.
Further, the radix tetrastigme zymolyte is prepared via a method which: it is dry by radix tetrastigme rhizome, powder is beaten,
Be added the deionized water of 10 times of weight, addition volume mass ratio is 0.1-0.2ml/100g amylase, 70 DEG C enzymatic hydrolysis 3-5 hours,
Obtain radix tetrastigme zymolyte.
In the above-mentioned technical solutions, the effect of big bean sprout slurry is: the V of the protein of germinated soybeanB2、VB6、VCEqual size
It significantly improves；The anti-nutritional factors such as trypsin inhibitor and phytic acid are also because germination is decomposed, isoflavones, lecithin, soap
Angle glycosides equal size also increases to some extent, so that the nutritive value and medical value of germinated soybean are all improved；
The effect of radix tetrastigme zymolyte is: using amylase to the enzymatic saccharification of much starch in radix tetrastigme, hydrolysis life
As the maltose and glucose for being easy to be absorbed by strain, discharge effective component preferably.
The effect of ramulus mori and blue or green money willow extract is: mycelial biomass can be improved.
The effect of lactose is: can promote the synthesis of polysaccharide.
Calcium chloride and salicylic effect are: can promote the synthesis of triterpene compound.
A kind of fermentation culture method preparing high activity Phellinus tunning, includes the following steps:
(1) the Phellinus strain after activation is inoculated in seed fluid nutrient mediums of saccharomycete and is carried out Phellinus fermented and cultured 3-7 days, obtained
Phellinus liquid seeds liquid；
(2) volume that first access accounts for the fermentation medium into any fermentation medium of above scheme is 1%-
The 5% Phellinus liquid seeds liquid, is cultivated 5-8 days, fermentation ends under the conditions of 24-27 DEG C of temperature, and fermentation liquid is cold through vacuum
It is lyophilized dry, obtains high activity Phellinus tunning.
Further, step (1) described seed fluid nutrient mediums of saccharomycete by weight, is grouped as: germinated soybean by following group
20 parts, 5 parts of glucose, 0.05 part of magnesium sulfate, 0.02 part of potassium dihydrogen phosphate.
Further, step (1) the Phellinus strain is selected from the I.sanghuang of Hangzhou, Zhejiang province city Thousand-Island Lake.
The beneficial effects of the present invention are:
(1) pass through the nutritional ingredient fermented by fermentation culture and effective component preferably carries out biodegrade and turns
Change, helps to improve the content and activity of active constituent in final products Phellinus tunning.
(2) the high activity Phellinus tunning prepared has the subacute oxidative damage of D- galactolipin inducing mouse apparent
Protective effect.
Detailed description of the invention
Fig. 1 is influence diagram of the Phellinus tunning to mice serum oxidation index MDA.
Fig. 2 is influence diagram of the Phellinus tunning to mice serum oxidation index T-AOC.
Specific embodiment
Below by specific embodiment, the present invention is described in detail, but the purposes of these exemplary embodiments and
Purpose is only used to enumerate the present invention, not constitutes any type of any restriction to real protection scope of the invention, more non-to incite somebody to action
Protection scope of the present invention is confined to this.
Embodiment 1
The method for preparing high activity Phellinus tunning, sequentially includes the following steps:
(1) configure fermentation medium, the composition of fermentation medium: big bean sprout starches 20g/L, Semen Tritici aestivi fiber element 5g/L, three leaves
Green zymolyte 5g/L, ramulus mori and blue or green money willow extract 2g/L, calcium chloride 0.05g/L, lactose 0.3g/L, glucose 1g/L, bigcatkin willow
Sour 0.02g/L, water 80g/L.
Wherein, the preparation of big bean sprout slurry: selecting the complete soybean of grain shape, be put into Full-automatic bean sprout machine and spray vernalization, when
When soybean bud head is revealed to 4-5mm, stop germination；By big bean sprout: water quality ratio 1:1 is beaten at a high speed spare.
The preparation of ramulus mori and blue or green money willow extract: respectively crushing ramulus mori and Qingqian Willow leaf, mixed by weight the ratio of 1:1
It closes, concussion in 40-50 DEG C of ethanol solution of the shaking table of the 70vol% of 20-30 times of amount (weight) is added and extracts 10 hours, after filtering
It obtains supernatant and obtains extract after vacuum freeze drying after supernatant is concentrated under reduced pressure.
Radix tetrastigme zymolyte: it is dry by radix tetrastigme rhizome, powder is beaten, the deionized water of 10 times of amounts is added, volume mass is added
Than for (0.1-0.2ml/100g) amylase, 70 DEG C enzymatic hydrolysis 3-5 hours, it is spare.
(2) Phellinus liquid fermentation and culture；Phellinus strain after activation is inoculated in seed fluid nutrient mediums of saccharomycete and carries out Phellinus
Fermented and cultured 3-7 days, obtain Phellinus liquid seeds liquid；The wherein composition of seed fluid nutrient mediums of saccharomycete are as follows: contain hair in 1L in fermentation liquid
Bud soybean 20g, glucose 5g, magnesium sulfate 0.05g, potassium dihydrogen phosphate 0.02g.And selected Phellinus strain is selected from as follows:
| Collecting location/strain source | Host/culturing raw material | ITS comparison result | GenBank serial number |
| Thousand-Island Lake, Hangzhou, Zhejiang Province | Mulberry tree | I.sanghuang | HM584808 |
Remarks: ITS (Internal Transcribed Spacer) identification, which refers to, carries out DNA sequencing to ITS sequence, leads to
The ITS sequence for obtaining sequencing is crossed compared with fungi known ITS sequence, to obtain a kind of side of unknown fungi kind information
Method.
(3) liquid fermentation and culture: first access accounts for fermentation medium volume 1%- in the fermentation medium in step (1)
5% Phellinus liquid seeds liquid is cultivated 5-8 days, fermentation ends under the conditions of 24-27 DEG C of temperature, and fermentation liquid is dry through vacuum refrigeration
It is dry, obtain high activity Phellinus tunning.
Embodiment 2
Referring to embodiment 1, unlike the first embodiment, the composition of fermentation medium: big bean sprout starches 25g/L, Semen Tritici aestivi fiber
Plain 7g/L, radix tetrastigme zymolyte 8g/L, ramulus mori and blue or green money willow extract 3g/L, calcium chloride 0.08g/L, lactose 0.4g/L, grape
Sugared 2g/L, salicylic acid 0.04g/L, water 90g/L.
Embodiment 3
Referring to embodiment 1, unlike the first embodiment, the composition of fermentation medium: big bean sprout starches 30g/L, Semen Tritici aestivi fiber
Plain 10g/L, radix tetrastigme zymolyte 10g/L, ramulus mori and blue or green money willow extract 3g/L, calcium chloride 0.1g/L, lactose 0.5g/L, grape
Sugared 3g/L, salicylic acid 0.05g/L, water 100g/L.
Comparative example 1
Referring to embodiment 2, unlike the first embodiment, in the composition of fermentation medium, by ramulus mori and blue or green money willow extract
It is changed to extract of mulberry twig.
The preparation of extract of mulberry twig: ramulus mori is crushed, and the ethanol solution 40- of the 70vol% of 20-30 times of amount (weight) is added
Concussion is extracted 10 hours in 50 DEG C of shaking table, and supernatant is obtained after filtering, after supernatant is concentrated under reduced pressure, after vacuum freeze drying
To extract.
Comparative example 2
Referring to embodiment 2, unlike the first embodiment, in the composition of fermentation medium, by ramulus mori and blue or green money willow extract
It is changed to blue or green money willow extract.
The preparation of extract of mulberry twig: Qingqian Willow leaf is crushed, and the ethyl alcohol that the 70vol% of 20-30 times of amount (weight) is added is molten
Concussion is extracted 10 hours in 40-50 DEG C of liquid of shaking table, and supernatant is obtained after filtering, and after supernatant is concentrated under reduced pressure, vacuum refrigeration is dry
Extract is obtained after dry.
The calculating that mycelia yield is carried out to the Phellinus tunning that embodiment 2, comparative example 1-2 are obtained, referring to table 1.
Table 1
| Mycelia yield | |
| Embodiment 2 | 4.8% |
| Comparative example 1 | 4.0% |
| Comparative example 2 | 4.1% |
Embodiment 5
The high activity Phellinus tunning that embodiment 2 obtains is tested as follows:
Will be spare after Phellinus tunning (rear abbreviation fermentation material) drying and crushing of embodiment 2,60 ICR mouse are divided at random
For Normal group, model group, positive drug group, fermentation material high, medium and low dosage group (18.0g/kg, 12.0g/kg, 6.0g/
Kg), every group 10, in addition to Normal group, remaining 5 groups prepare the subacute peroxidating of mouse using D- galactolipin hypodermic injection
Damage model, i.e., the D- galactolipin of the daily subcutaneous multi-point injection 120mg/kg of timing the nape of the neck, Normal group are raw with injection
It manages salt water to replace, continuous 42d.(next day starts gastric infusion after injection of d-galactose for the first time, by 20ml/kg while modeling
Weight stomach-filling), fermentation material each group stomach-filling corresponding dosage, positive controls give vitamin E 200mg/kg, Normal group and
Model group gives drinking water.Fasting for 24 hours, plucks eyeball and takes blood, take liver and brain after the last administration.It is quantitative determined and is tried using total protein
Agent box measures liver protein concentration.The superoxide dismutase in method measurement blood and hepatic tissue illustrated according to kit
(SOD), the content of glutathione peroxidase (GSH), total antioxidant capacity (T-AOC) and malonaldehyde (MDA).Test result
Referring to shown in table 2 and Fig. 1 and Fig. 2.
2 fermentation material of table is to murine liver tissue T-AOC, the influence of GSH and SOD content
As depicted in figs. 1 and 2, compared with Normal group, MDA is significantly increased in model group serum, and T-AOC significantly drops
Low, each dosage group MDA content of fermentation material significantly reduces, and T-AOC is without significant change.Positive group MDA and T-AOC is poor without statistics
It is different.Compared with model group, MDA content is significantly reduced in each dosage group of fermentation material and positive group serum, and T-AOC is horizontal significantly to be risen
It is high.
As shown in table 2, compared with Normal group, GSH is horizontal in model group and each dosage group murine liver tissue of fermentation material
Without significant change, downward trend is presented in model group T-AOC and SOD enzyme activity.Fermentation material 6.0 and 12.0g/kg dosage group SOD without
Significant change, the decline of fermentation material high dose group SOD vigor.Obvious rising is presented in positive group T-AOC and GSH, and SOD enzyme activity is presented
The trend of liter, but no difference of science of statistics.Compared with model group, positive group GSH and SOD vigor is significantly increased, and T-AOC is increased, but nothing
Statistical difference.GSH level is omited without significant change, fermentation material high dose group T-AOC in each dosage group murine liver tissue of fermentation material
There is rising, SOD enzyme activity significantly increases, and has statistical difference (P < 0.05).
Therefore, Phellinus tunning is under 6.0,12.0 and 18.0g/kg dosage to the subacute oxygen of D- galactolipin inducing mouse
Changing damage has significant protective effect.
The series of detailed descriptions listed above are illustrated only for possible embodiments of the invention,
The protection scope that they are not intended to limit the invention, it is all without departing from equivalent embodiment made by technical spirit of the present invention or change
It should all be included in the protection scope of the present invention.
Claims (7)
1. a kind of fermentation medium for being used to prepare high activity Phellinus tunning, which is characterized in that by weight, by as follows
Group is grouped as: big bean sprout starches 20-30 parts, and 5-10 parts of Semen Tritici aestivi fiber element, 5-10 parts of radix tetrastigme zymolyte, ramulus mori and blue or green money willow are extracted
2-3 parts of object, 0.05-0.1 parts of calcium chloride, 0.3-0.5 parts of lactose, 1-3 parts of glucose, 0.02-0.05 parts of salicylic acid, water 80-100
Part.
2. the fermentation medium according to claim 1 for being used to prepare high activity Phellinus tunning, which is characterized in that institute
It states big bean sprout slurry to be prepared via a method which: selecting the complete soybean of grain shape, be put into Full-automatic bean sprout machine and spray vernalization,
When soybean bud head is revealed to 4-5mm, stop germination；Big bean sprout: water is obtained into big bean sprout by weight 1: 1 mixing, high speed mashing
Slurry.
3. the fermentation medium according to claim 1 for being used to prepare high activity Phellinus tunning, which is characterized in that institute
It states ramulus mori and blue or green money willow extract is prepared via a method which: respectively crushing ramulus mori and Qingqian Willow leaf, by weight 1:1
Ratio mixing, is added the ethanol solution of the 70vol% of 20-30 times of weight, and concussion is extracted 10 hours in 40-50 DEG C of shaking table,
Supernatant is obtained after filtering after vacuum freeze drying, obtains ramulus mori and blue or green money willow extract after supernatant is concentrated under reduced pressure.
4. the fermentation medium according to claim 1 for being used to prepare high activity Phellinus tunning, which is characterized in that institute
Radix tetrastigme zymolyte is stated to be prepared via a method which: it is dry by radix tetrastigme rhizome, beat powder, be added 10 times of weight go from
Sub- water, addition volume mass ratio be 0.1-0.2ml/100g amylase, 70 DEG C enzymatic hydrolysis 3-5 hours, obtain radix tetrastigme zymolyte.
5. a kind of fermentation culture method for preparing high activity Phellinus tunning, which comprises the steps of:
(1) the Phellinus strain after activation is inoculated in seed fluid nutrient mediums of saccharomycete and is carried out Phellinus fermented and cultured 3-7 days, obtain Phellinus
Liquid seeds liquid；
(2) volume that first access accounts for the fermentation medium into any fermentation medium of claim 1-4 is 1%-
The 5% Phellinus liquid seeds liquid, is cultivated 5-8 days, fermentation ends under the conditions of 24-27 DEG C of temperature, and fermentation liquid is cold through vacuum
It is lyophilized dry, obtains high activity Phellinus tunning.
6. the fermentation culture method of preparation high activity Phellinus tunning according to claim 5, which is characterized in that step
(1) seed fluid nutrient mediums of saccharomycete by weight, is grouped as by following group: 20 parts of germinated soybean, 5 parts of glucose, and magnesium sulfate
0.05 part, 0.02 part of potassium dihydrogen phosphate.
7. the fermentation culture method of preparation high activity Phellinus tunning according to claim 5, which is characterized in that step
(1) the Phellinus strain is selected from the I.sanghuang of Hangzhou, Zhejiang province city Thousand-Island Lake.
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Cited By (3)
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| CN110904156A (en) * | 2019-12-23 | 2020-03-24 | 江南大学 | Method for increasing yield of triterpenoids in phellinus igniarius liquid state fermentation |
| CN112063663A (en) * | 2020-08-24 | 2020-12-11 | 浙江省农业科学院 | Fermentation method for high-yield phellinus igniarius extracellular flavonoids |
| 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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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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