CN105671100B - A method of synthesis Phellinus high activity flavone compound - Google Patents
A method of synthesis Phellinus high activity flavone compound Download PDFInfo
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Abstract
The invention belongs to bio-fermentation engineering fields; more particularly to a kind of method for synthesizing Phellinus high activity flavone compound; mainly its culture medium is optimized; it is added to the substances such as cumarin, phenylalanine in the medium; four steps of Phellinus activity flavone compound are extracted by the culture of (1) first order seed, (2) secondary seed culture, (3) fermented and cultured, (4) again; the flavone compound yield for substantially increasing Phellinus, also maintains its bioactivity.High yield and the active flavonoids from phellinus class compound of high anti-oxidation can be obtained simultaneously with the present invention, content is high, and activity stabilized, with short production cycle, pollution is few, and it is low in cost, it is a kind of great production method for having industrial prospect.
Description
Technical field
The invention belongs to biological fermentation engineering fields, are related to a kind of liquid for efficiently synthesizing Phellinus activity flavone compound
Fermentation process.
Background technique
Phellinus (Phellinus igniarius), belongs to Basidiomycotina (Basidiomycota), Hymenomycetes
(Hymenomycetes), Aphyllophorales (Polyporales), Polyporaceae (Hymenochatetacae), Phellinus
(Phellinus) rare medicinal fungi, also known as phelliuns igniarius (P.igniarius).Phellinus is a kind of traditional Chinese medicine, taste
Slight bitter, energy relieving the five internal organs, softening hard masses, toxin expelling, hemostasis, promoting blood circulation stomach function regulating antidiarrheal are civil common to treat stranguria syndrome, metrorrhagia and metrostaxis leukorrhagia, sore cave product
Poly-, weakness, splenasthenic diarrhea etc..The latest study proves Phellinus has anti-oxidant, antibacterial anti-inflammatory, a series of pharmacology such as antitumor are living
Property, and the active constituent in Phellinus is mainly flavone compound and polysaccharide.Wherein flavone compound has antioxidation,
It is antitumor, it is hypoglycemic, the effects of immunological regulation.However due to being limited by various environmental conditions, Phellinus shape in nature
At fructification it is rare, flavone compound is extracted from its fructification, low yield, activity is not high and extremely difficult, because
This, is synthesized utilization and exploitation with Phellinus by the flavone compound that biological engineering method efficiently synthesizes high activity and high yield
Flavonoid substances using extremely important.
Liu Fan et al. discloses a kind of liquid fermentation method for improving phellinus igniarius mycelium yield of flavone, and (application publication number is
CN103695316A), yield can reach 0.6g/L, and Zhu Hu et al. discloses a kind of technique of liquid-solid two-phase culture flavonoids from phellinus
(application publication number CN103627728A), the maximum output for obtaining flavones is 2.174g/L.Happiness happiness et al. is lured using fungi
Guide improves yield of flavone (application publication number CN101702984A) in Phellinus liquid fermentation, obtains the maximum output of flavones
It is 0.34g/L, currently, also reporting many documents to the optimization of Phellinus fermentation medium at home, Zhao Zigao et al. passes through excellent
The yield that culture medium after change obtains flavones is 0.128056g/L, and Xu Qian et al. obtains flavones by the culture medium after optimization
Yield is 0.21235g/L, these all provide method and fermentation medium to improve flavonoids from phellinus class compound production, still
Their yield is all very low, does not also all ensure the bioactivity of flavone compound in the synthesis process.Using in the present invention
Formula synthesize such reactive compound, the up to flavone compound of 5.9-6.5g/L can be obtained, and antioxidant activity is
5.1-6.7mmol/L, this is significantly larger than above-mentioned patent from yield or document is reported or the numerical value of publicity method, and first
It is secondary to ensure the bioactivity for synthesizing such compound in the synthesis process, it is primarily due in the research of this formulation optimization, removes
Consider outside influence of the single factor test to Phellinus synthesis flavone compound yield, also from meeting Phellinus synthesis high yield and high activity
The angle of flavone compound is set out, be added to cumarin etc. to ensure culture medium that active group in synthesis process is formed at
Point, and synthesized from the point of view of flavone compound yield and bioactivity between medium component from Phellinus is influenced simultaneously
Reciprocation forms while improving the yield of flavonoids from phellinus class compound and the culture medium condition of bioactivity, this is current
It is unexistent in the document or patent of report.
Although the technical system of flavone compound is synthesized there are many now using fungi or plant cell, they
All there is following defect:
1, the yield of flavone compound is only focused in the synthesis process, and has ignored the formation problem of its bioactivity,
This is unfavorable for the flavone compound for obtaining high activity.
2, the ingredient in culture medium prescription reported at present is complex, mostly using natural products as carbon source and nitrogen source, this
So that there are obvious batch wise differences for the quality and activity of production process, be unfavorable for flavone compound stablizes synthesis.
3, the finally obtained flavone compound yield of these methods is not high, and bioactivity is not guaranteed.
It would therefore be desirable to find a kind of method for efficiently synthesizing active flavone compound, the present invention then passes through science
Statistical method-response surface optimization has researched and solved the above problem in Phellinus fermentation medium.Not only synthesis process is easy to operate,
It is low in cost, moreover it is possible to which that verifying can synthesize the flavone compound for having high activity in production level scale with high yield；Experiment
The verified flavone compound in room has clearly hypoglycemic, anti-oxidant, anti-aging and anti-trioxypurine effect, is to prepare this
The health-related drink and food of class, dietary supplements, food additives and related drugs important raw materials for production.
Summary of the invention
It is an object of the invention to overcome defect present in the prior art, provide it is a kind of easy to operate, it is low in cost,
The method for having the flavone compound of high activity can be synthesized in production level high yield.
To achieve the goals above, the technical solution adopted by the present invention is made of following step:
(1) culture of first order seed
The activated Phellinus strain of slant preservation is inoculated into progress first order seed culture, inoculum concentration in seed culture medium
It is the fungus block that 4~5 block sizes of access are 5mm × 5mm in the seed culture medium of every 100ml, cultivation temperature is 27~30 DEG C, revolving speed
For 150~180r/min, incubation time is 7~9 days；
In above-mentioned 1L seed culture medium: glucose is 15.0~22.0g, potato is 150.0~220.0g, KH2PO4For
0.5~2.5g, MgSO4For 0.2~0.5g, 1L is added water to, the pH of the seed culture medium is 6.5；
(2) culture of secondary seed
Cultured first order seed in step (1) is connected to progress secondary seed culture, inoculum concentration in seed culture medium
For 5~15 (V/V) %, cultivation temperature is 27~30 DEG C, and revolving speed is 150~180r/min, and incubation time is 7~9 days, is trained
Feeding secondary seed；
(3) fermented and cultured
The cultured secondary seed of step (2) is connected in fermentation medium, inoculum concentration is 5~15 (V/V) %, culture
Temperature is 27~30 DEG C, and revolving speed is 150~180r/min, cultivates 4~5 days and D-101 macroreticular resin is added into fermentation medium,
Continue culture 3~4 days；
In 1L fermentation medium: glucose is 15.0~22.0g, peptone is 2.0~7.0g, cumarin be 0.02~
0.06g, phenylalanine are 0.2~0.6g, Tween-80 is 1.0~4.0g, 0.05~0.5g of cinnamic acid, MgSO4For 1.0~
5.0g、KH2PO4For 0.5~3.0g, 1L is added water to, the pH of the fermentation medium is 6.5；
(4) Phellinus activity flavone compound is extracted
D-101 macroreticular resin is filtered out after the completion of the fermented and cultured of step (3), will be fermented using D-101 macroreticular resin
Flavone compound in culture medium adsorbs, and is eluted out with 80% methanol, obtains Phellinus activity flavone compound.
The entitled Phellinus sp.P0988 of Phellinus strain in above-mentioned steps (1), deposit number are CCTCC NO:
M2012080, depositary institution are China typical culture collection centers, and address is Wuhan, China Wuhan University, postcode 430012,
Preservation date is on March 14th, 2012.
The optimization formula of above-mentioned steps (3) is: in 1L fermentation medium glucose be 18.0~20.0g, peptone 3.0
~5.0g, cumarin are 0.02~0.04g, phenylalanine is 0.25~0.5g, cinnamic acid is 0.05~0.3g, MgSO4It is 1.0
~3.0g, KH2PO4It is 1.0~3.0g for 0.5~2.0g, Tween-80, adds water to 1L, the pH of the fermentation medium is 6.5.
The method that the present invention synthesizes Phellinus high activity flavone compound, mainly in the culture medium to Phellinus fermentation synthesis
It is optimized, technical advantage is as follows:
(1) being had studied by response surface design optimization method, which influences Phellinus, synthesizes flavone compound yield and bioactivity
Medium component and its existing reciprocation between each other.It has been determined on the basis of statistical analysis and has significantly affected Phellinus
Synthesize the interaction between flavone compound yield and active important medium component cumarin, phenylalanine and Tween-80
Effect and primary influences, so that it is determined that these medium components synthesize flavone compound yield and activity most beneficial for Phellinus
The concentration range of formation.This is never by publicity or to report in the synthesis of flavone compound.
(2) skeleton of flavone compound is formed by joined cumarin, phenylalanine etc. in its fermentation medium
Precursor substance reduces the metabolic process that Phellinus synthesizes the substance, shortens the production cycle, promotes Phellinus synthesis process
In high activity forming process.
(3) present invention is also added into cinnamic acid, as the stimulant of secondary metabolite, promotes the synthesis and activity of flavones
Forming process.
(4) this method is easy to operate, low in cost, can get the flavone compound of high yield high activity, in food and
Pharmaceutical technology field is widely used, and therefore, the present invention is of great significance for industrialized production flavone compound
Specific embodiment
It is further illustrated below with reference to experiment.
Embodiment 1
The method of the present embodiment synthesis Phellinus high activity flavone compound comprises the steps of:
(1) first order seed culture
The activated Phellinus strain of slant preservation is inoculated into progress first order seed culture, inoculum concentration in seed culture medium
Be every 100ml seed culture medium in access 4 block sizes be 5mm × 5mm fungus block, 28 DEG C of cultivation temperature, revolving speed 160r/min,
Incubation time 9 days.
Phellinus strain is conventionally to activate, and specifically Phellinus strain is seeded in PDA culture medium, 25~30
DEG C, aerobic culture to the Phellinus strain that is protected from light covers with inclined-plane；The entitled Phellinus sp.P0988 of the Phellinus strain, preservation list
Position is China typical culture collection center, and address is Wuhan, China Wuhan University, postcode 430012, deposit number CCTCC
NO:M 2012080, preservation date are on March 14th, 2012.
In above-mentioned 1L seed culture medium: glucose 20.0g, potato 200.0g, KH2PO4 1g、MgSO40.5g, surplus
For water, the pH of the seed culture medium is 6.5.
(2) secondary seed culture
Cultured first order seed in step (1) is connected to progress secondary seed culture, inoculum concentration in seed culture medium
For 10 (V/V) %, i.e., the first order seed of 10mL is inoculated in the seed culture medium of every 100mL, cultivation temperature is 28 DEG C, and revolving speed is
160r/min, incubation time are 9 days, obtained secondary seed.
(3) fermented and cultured
Cultured secondary seed in step (2) is connected in fermentation medium, inoculum concentration is 10 (V/V) %, i.e., often
The secondary seed of 10mL is inoculated in the fermentation medium of 100mL, cultivation temperature is 28 DEG C, revolving speed 160r/min, it cultivates 4 days,
D-101 macroreticular resin is added into fermentation medium to be adsorbed, culture 4 days is continued；
In above-mentioned 1L fermentation medium: glucose 20.0g, peptone 4g, cumarin 0.03g, phenylalanine 0.35g,
Cinnamic acid 0.1g, MgSO4 1.5g、KH2PO41.0g, Tween-80 2.0g, add water to 1L, and the pH of the fermentation medium is 6.5.
(4) Phellinus activity flavone compound is extracted
D-101 macroreticular resin is filtered out after the completion of the fermented and cultured of step (3), will be fermented using D-101 macroreticular resin
The flavone compound generated in culture medium adsorbs, and is eluted out with 80% methanol, obtains Phellinus activity flavonoids
The content closed object, and determine flavone compound is 5.76g/L, total antioxidant activity 5.15mmol/L.
Embodiment 2
The method of the present embodiment synthesis Phellinus high activity flavone compound comprises the steps of:
(1) first order seed culture
The activated Phellinus strain of slant preservation is inoculated into progress first order seed culture, inoculum concentration in seed culture medium
Be every 100ml seed culture medium in access 5 block sizes be 5mm × 5mm fungus block, 27 DEG C of cultivation temperature, revolving speed 180r/min,
Incubation time 8 days；
In above-mentioned 1L seed culture medium: glucose 18.0g, potato 180.0g, KH2PO4 1g、MgSO40.5g adds water
To 1L, the pH of the seed culture medium is 6.5.
(2) secondary seed culture
Cultured first order seed in step (1) is connected to progress secondary seed culture, inoculum concentration in seed culture medium
For 15 (V/V) %, i.e., the first order seed of 15mL is inoculated in the seed culture medium of every 100mL, cultivation temperature is 27 DEG C, and revolving speed is
180r/min, incubation time are 8 days, obtain secondary seed.
(3) fermented and cultured
Cultured secondary seed in step (2) is connected in fermentation medium, inoculum concentration is 15 (V/V) %, i.e., often
The secondary seed of 15mL is inoculated in the fermentation medium of 100mL, cultivation temperature is 27 DEG C, revolving speed 150r/min, it cultivates 5 days,
D-101 macroreticular resin is added into fermentation medium to be adsorbed, culture 3 days is continued；
In above-mentioned 1L fermentation medium: glucose 18.0g, peptone 3g, cumarin 0.04g, phenylalanine 0.4g, meat
Cinnamic acid 0.3g, MgSO4 4.0g、KH2PO40.5g, Tween-80 1.0g, add water to 1L, and the pH of the fermentation medium is 6.5.
(4) Phellinus activity flavone compound is extracted
D-101 macroreticular resin is filtered out after the completion of the fermented and cultured of step (3), will be fermented using D-101 macroreticular resin
The flavone compound generated in culture medium adsorbs, and is eluted out with 80% methanol, obtains Phellinus activity flavonoids
The content closed object, and determine flavone compound is 4.054g/L, total antioxidant activity 5.1964mmol/L.
Embodiment 3
The method of the present embodiment synthesis Phellinus high activity flavone compound comprises the steps of:
(1) first order seed culture
The activated Phellinus strain of slant preservation is inoculated into progress first order seed culture, inoculum concentration in seed culture medium
Be every 100ml seed culture medium in access 5 block sizes be 5mm × 5mm fungus block, 30 DEG C of cultivation temperature, revolving speed 150r/min,
Incubation time 9 days；
In above-mentioned 1L seed culture medium: glucose 22.0g, potato 150.0g, KH2PO4 0.5g、MgSO40.5g adds
For water to 1L, the pH of the seed culture medium is 6.5.
(2) secondary seed culture
Cultured first order seed in step (1) is connected to progress secondary seed culture, inoculum concentration in seed culture medium
For 5 (V/V) %, i.e., the first order seed of 5mL is inoculated in the seed culture medium of every 100mL, cultivation temperature is 30 DEG C, and revolving speed is
150r/min, incubation time are 9 days, obtain secondary seed.
(3) fermented and cultured
Cultured secondary seed in step (2) is connected in fermentation medium, inoculum concentration is 5 (V/V) %, i.e., often
The secondary seed of 5mL is inoculated in the fermentation medium of 100mL, cultivation temperature is 30 DEG C, revolving speed 180r/min, it cultivates 4 days, to
D-101 macroreticular resin is added in fermentation medium to be adsorbed, culture 3 days is continued；
In above-mentioned 1L fermentation medium: glucose 20.0g, peptone 5g, cumarin 0.02g, phenylalanine 0.5g, meat
Cinnamic acid 0.05g, MgSO4 1.0g、KH2PO42.0g, Tween-80 3.0g, add water to 1L, and the pH of the fermentation medium is 6.5.
(4) Phellinus activity flavone compound is extracted
D-101 macroreticular resin is filtered out after the completion of the fermented and cultured of step (3), will be fermented using D-101 macroreticular resin
The flavone compound generated in culture medium adsorbs, and is eluted out with 80% methanol, obtains Phellinus activity flavonoids
Object is closed, and determining its content is 4.73g/L, total antioxidant activity 6.4mmol/L.
Embodiment 4
In the present embodiment, the seed culture based formulas of step (1) is: glucose 15.0g, potato 220.0g, KH2PO4
2.5g、MgSO40.2g adds water to 1L, and the pH of the seed culture medium is 6.5.
The formula of fermentation medium used is in step (3): glucose 15.0g, peptone 2g, cumarin 0.06g, benzene
Alanine 0.6g, cinnamic acid 0.5g, MgSO4 5.0g、KH2PO43.0g, Tween-80 4.0g, add water to 1L, the fermented and cultured
The pH of base is 6.5.
Others operation is same as Example 1, and yield and total antioxidant activity are close with the result of embodiment 1.
Embodiment 5
In the present embodiment, the seed culture based formulas of step (1) is: glucose 22.0g, potato 150.0g, KH2PO4
2.0g、MgSO40.3g adds water to 1L, and the pH of the seed culture medium is 6.5.
The formula of fermentation medium used is in step (3): glucose 22.0g, peptone 7.0g, cumarin 0.02g, benzene
Alanine 0.2g, cinnamic acid 0.05g, MgSO4 1.0g、KH2PO40.5g, Tween-80 1.0g, add water to 1L, the fermented and cultured
The pH of base is 6.5.
Others operation is same as Example 1, and yield and total antioxidant activity are close with the result of embodiment 1.
In order to verify technical effect of the invention, applicant has done a large amount of experiment, is now said by taking following experiments as an example
It is bright.
(1) fermentation medium compares
In the fermentation medium of comparative example 1: glucose 20.0g, peptone 4g, KH2PO4 1.0g、MgSO41.5g, cortex cinnamomi
Sour 0.1g, pH 6.5.
In the fermentation medium of comparative example 2: glucose 20.0g, peptone 4g, KH2PO4 1.0g、MgSO41.5g, cortex cinnamomi
Sour 0.1g, cumarin 0.03g, pH 6.5.
In the fermentation medium of comparative example 3: glucose 20.0g, peptone 4g, KH2PO4 1.0g、MgSO41.5g, cortex cinnamomi
Sour 0.1g, phenylalanine 0.35g, pH 6.5.
In the fermentation medium of comparative example 4: glucose 20.0g, peptone 4g, KH2PO4 1.0g、MgSO41.5g, cortex cinnamomi
Sour 0.1g, Tween-80 2.0g, pH 6.5.
Other the step of, are same as Example 1, experimental results such as the following table 1:
Table 1 is the Comparative result of each comparative example and embodiment 1
| Flavone compound | Embodiment 1 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 |
| Yield | 5.76g/L | 1.90g/L | 2.63g/L | 2.95g/L | 3.35g/L |
| Total antioxidant activity | 5.15mmol/L | 0.74mmol/L | 1.44mmol/L | 3.64mmol/L | 1.45mmol/L |
As can be seen from Table 1, when being separately added into cumarin, phenylalanine and Tween-80, flavonoids can be improved
Compound production and activity, but effect is not obvious, and three ought be added simultaneously, then substantially increase the production of flavone compound
Amount and activity, this sufficiently proves that cumarin, phenylalanine and Tween-80 have synergistic effect, and this three is to flavonoids
The synthesis for closing object has facilitation.
For the relationship for verifying three in this experiment, analyzed in conjunction with following experiment:
The design principle of Central Composite experiment in the method for face according to response, uses X respectively1、X2、X3To represent cumarin, phenylpropyl alcohol
Propylhomoserin and Tween-80, then encoded respectively with -1,0,1 by the basic, normal, high level of this 3 independents variable.Respectively with Phellinus Huang
Ketone compounds content and its total antioxidant activity are response contrived experiment, further select the allocation optimum of significant factor dense
Relationship between degree and analysis three, response surface experimental design and the results are shown in Table 2.
2 response surface experimental design of table and result
Using flavonoid content as response, according to CCD contrived experiment in upper table as a result, with Minitab software
Quadratic Regression Analysis is carried out to result therein, obtains quadratic regression equation:
Y=0.16458-0.1394X1+0.0406X2+0.1374X3+0.24055X1 2+0.40255X2 2+1.29355X3 2+
0.26913X1X2+0.43137X1X3+1.58812X2X3
The variance analysis of regression equation is shown in Table 3
The variance analysis of 3 regression equation of table
Using the total antioxidant activity of flavone compound as response, according to the experimental result that CCD in table 2 is designed, use
Minitab software carries out Quadratic Regression Analysis to it, obtains quadratic regression equation and is
Y=0.775-0.224X1+0.124X2-0.086X3+0.48X1 2+0.52X2 2+1.42X3 2+0.37X1X2+0.55
X1X3+1.68X2X3
The variance analysis of regression equation is shown in Table 4
The variance analysis of 4 regression equation of table
The reciprocation highly significant (P < 0.001) of equation it can be seen from table 3 and table 4, and equation is linear, flat
Square item is also very significant, illustrates that the influence of each experiment factor pair flavone compound is not simple linear relationship, they are to Huang
The yield and antioxidant activity of ketone compounds all have an impact, and further illustrate and have between cumarin, phenylalanine and Tween-80
There is mutually coordinated, the relationship of interaction.
(2) influence of the additive amount of cumarin to flavone compound
In the fermentation medium of comparative example 4: glucose 20.0g, peptone 4g, cumarin 0.1g, phenylalanine 1.0g,
MgSO4 1.5g、KH2PO41.0g, Tween-80 2.0g, pH 6.5.
In the fermentation medium of comparative example 5: glucose 20.0g, peptone 4g, cumarin 0.005g, phenylalanine
0.05g、MgSO4 1.5g、KH2PO41.0g, Tween-80 2.0g, pH 6.5.
Other the step of, are same as Example 1, experimental results such as the following table 5:
Table 5 is the Comparative result of each comparative example and embodiment 1
| Flavone compound | Embodiment 1 | Comparative example 4 | Comparative example 5 |
| Yield | 5.76g/L | 1.13g/L | 1.99g/L |
| Total antioxidant activity | 5.15mmol/L | 1.11mmol/L | 1.91mmol/L |
The cumarin of different content has not flavone compound yield and flavone compound activity as can be seen from Table 5
Same influence illustrates tonka-bean more than when the content of cumarin, the content and activity of flavone compound can be reduced in this formula
The excessively high synthesis that can inhibit flavone compound of cellulose content, and lower than in this formula when the content of cumarin, flavone compound
Content and activity can also reduce, this illustrates that tonka-bean cellulose content is too low, with other factors interaction effect it is weaker, be unfavorable for
The synthesis of flavone compound.
Claims (1)
1. a kind of method for synthesizing Phellinus high activity flavone compound, it is characterised in that be made of following step:
(1) culture of first order seed
The activated Phellinus strain of slant preservation is inoculated into progress first order seed culture in seed culture medium, inoculum concentration is every
The fungus block that 4~5 block sizes are 5mm × 5mm is accessed in the seed culture medium of 100 ml, cultivation temperature is 27~30 DEG C, and revolving speed is
150~180 r/min, incubation time are 7~9 days；
The Phellinus strain it is entitledPhellinusSp.P0988, deposit number are CCTCC NO:M2012080, preservation list
Position is China typical culture collection center, and address is Wuhan, China Wuhan University, and preservation date is on March 14th, 2012；
In above-mentioned seed culture medium: glucose is 15.0~22.0g, potato is 150.0~220.0g, KH2PO4 For 0.5~
2.5g、MgSO4For 0.2~0.5g, 1L is added water to, the pH of the seed culture medium is 6.5；
(2) culture of secondary seed
Cultured first order seed in step (1) is connected in seed culture medium progress secondary seed culture, inoculum concentration is 5~
15 (V/V) %, cultivation temperature are 27~30 DEG C, and revolving speed is 150~180r/min, and incubation time is 7~9 days, obtain second level kind
Son；
(3) fermented and cultured
The cultured secondary seed of step (2) is connected in fermentation medium, inoculum concentration is 5~15 (V/V) %, cultivation temperature
It is 27~30 DEG C, revolving speed is 150~180r/min, cultivates 4~5 days and D-101 macroreticular resin is added into fermentation medium, continue
Culture 3~4 days；
In 1L fermentation medium: glucose 20.0g, peptone 4g, cumarin 0.03g, phenylalanine 0.35g, cinnamic acid 0.1g,
MgSO4 1.5g、KH2PO41.0g, Tween-80 2.0g, add water to 1L, and the pH of the fermentation medium is 6.5；
Or in 1L fermentation medium: glucose 18.0g, peptone 3g, cumarin 0.04g, phenylalanine 0.4g, cinnamic acid
0.3g、MgSO4 4.0g、KH2PO40.5g, Tween-80 1.0g, add water to 1L, and the pH of the fermentation medium is 6.5；
Or in 1L fermentation medium: glucose 20.0g, peptone 5g, cumarin 0.02g, phenylalanine 0.5g, cinnamic acid
0.05g、MgSO4 1.0g、KH2PO42.0g, Tween-80 3.0g, add water to 1L, and the pH of the fermentation medium is 6.5；
Or in 1L fermentation medium: glucose 15.0g, peptone 2g, cumarin 0.06g, phenylalanine 0.6g, cinnamic acid
0.5g、MgSO4 5.0g、KH2PO43.0g, Tween-80 4.0g, add water to 1L, and the pH of the fermentation medium is 6.5；
Or in 1L fermentation medium: glucose 22.0g, peptone 7.0g, cumarin 0.02g, phenylalanine 0.2g, cinnamic acid
0.05g、MgSO4 1.0g、KH2PO40.5g, Tween-80 1.0g, add water to 1L, and the pH of the fermentation medium is 6.5；
(4) Phellinus activity flavone compound is extracted
D-101 macroreticular resin is filtered out after the completion of the fermented and cultured of step (3), using D-101 macroreticular resin by fermented and cultured
The flavone compound generated in base adsorbs, and is eluted out with 80% methanol, obtains Phellinus activity flavone compound.
Priority Applications (1)
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| CN101182550A (en) * | 2007-11-16 | 2008-05-21 | 上海市农业科学院 | Flavonoids from phellinus, method of producing the same and use |
| CN103695316A (en) * | 2013-12-04 | 2014-04-02 | 广东省农业科学院蚕业与农产品加工研究所 | Liquid fermentation method for increasing flavone yield of phellinus igniarius mycelium |
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| CN101182550A (en) * | 2007-11-16 | 2008-05-21 | 上海市农业科学院 | Flavonoids from phellinus, method of producing the same and use |
| CN103695316A (en) * | 2013-12-04 | 2014-04-02 | 广东省农业科学院蚕业与农产品加工研究所 | Liquid fermentation method for increasing flavone yield of phellinus igniarius mycelium |
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| 微生物合成黄酮类化合物的研究进展;陈坚等;《食品科学技术学报》;20150131;第33卷(第1期);第1-5页 * |
| 药用菌桑黄代谢黄酮的调控研究;刘伟;《中国优秀硕士学位论文全文数据库 农业科技辑》;20140515(第5期);D047-230 * |
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