CN106636215A - Method of increasing active component content of phellinus igniarius - Google Patents
Method of increasing active component content of phellinus igniarius Download PDFInfo
- Publication number
- CN106636215A CN106636215A CN201611197331.4A CN201611197331A CN106636215A CN 106636215 A CN106636215 A CN 106636215A CN 201611197331 A CN201611197331 A CN 201611197331A CN 106636215 A CN106636215 A CN 106636215A
- Authority
- CN
- China
- Prior art keywords
- phellinus
- active component
- component content
- polysaccharide
- content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method of increasing the active component content of phellinus igniarius. According to the method, a seed solution of phellinus igniarius is inoculated in a liquid culture medium to carry out fermentation; and the liquid culture medium contains a carbon source, which is maltose, and a nitrogen source, which is a yeast extract. The concentration of maltose in the liquid culture medium is 2 to 5%; and the concentration of the yeast extract in the liquid culture medium is 0.1 to 0.4%. The invention provides a method of increasing the active component content of phellinus igniarius. A liquid fermentation mode is adopted, and the fermentation conditions are optimized to increase the contents of polysaccharide and flavones.
Description
Technical field
The present invention relates to Chinese medicine, more particularly to a kind of method for improving Phellinus active component content.
Background technology
Phellinus is the rare medicinal fungi of China.Phellinus another name monkey eye, mulberry ear, Sang Chen etc., are a kind of the large-scale of preciousness
Medicinal fungi, its fructification can be used as medicine.Gain the name because parasitizing mulberry tree, Phellinus is distributed more widely in China, be mainly grown in China
, also there is growth on the ground such as North China, northeast, northwest and Sichuan, Yunnan in some East Asian countries such as Korea and Japan, is described as " gloomy
Woods gold ", it is few with wild fructification quantity, the characteristics of artificial cultivation difficulty is high.Phellinus is used in blood by traditional chinese medicine
Collapse, blood strangury, the disease such as amenorrhoea.Modern pharmacology research shows that in addition Phellinus, also resists in anticancer aspect effect is significant
The effects such as pneumonia, reducing blood lipid, anti-oxidant and protection liver.It is current medicine and food industry joint research and focus.
With the further investigation of Phellinus medical value, market increases year by year for the demand of Phellinus, causes to Phellinus
The irrational extraction of resources of fructification, it is thin for the protective awareness in the place of production, cause fructification and be difficult to be molded, resource increasingly depleted,
It is difficult to search out the trace of Phellinus in some areas of China, most of producing region is precarious.Artificial cultivation difficulty is larger,
The development and utilization of Phellinus also therefore suffers from limiting.
Liquid fermentation technology, solid culture technology and artificial cultivation technique are currently acquired phellinus igniarius mycelium and fructification master
The approach wanted.At present, because fructification large-scale artificial cultivation technique is not still very ripe, liquid fermentation is adopted on market mostly
Technology is obtaining Phellinus fructification or non-fructification product, and the research to active component is very few.
The content of the invention
Goal of the invention：For problems of the prior art, the invention provides a kind of Phellinus active component that improves contains
The method of amount, by the way of liquid fermentation, by being optimized to fermentation condition, improves the content of polysaccharide and flavones.
Technical scheme：The method for improving Phellinus active component content of the present invention, including：By the inoculation of Phellinus seed liquor
Fermented in fluid nutrient medium, containing carbon source and nitrogen source in described fluid nutrient medium, described carbon source is maltose, institute
The nitrogen source stated is yeast extract.
Concentration in the maltose liquid medium within is 2-6%, preferably 2-4%, more preferably 2%.
Concentration in the yeast extract liquid medium within be 0.1-0.4%, preferably 0.2-0.4%, more preferably
For 0.4%.
Also contain MgSO during fermentation in fluid nutrient medium4·7H2O 0.4-0.6 ‰, KH2PO44.5-5‰.Preferably, send out
Also contain MgSO during ferment in fluid nutrient medium4·7H2O 0.5 ‰, KH2PO4 4.6‰。
Above-mentioned concentration refers to mass fraction.
The preparation method of the Phellinus seed liquor is：PDA liquid medium, 27-29 will be inoculated in after Phellinus actication of culture
6-7d is cultivated with 150-170r/min in DEG C shaking table.
During fermentation, inoculum concentration is 9-10%.
The temperature of the fermentation is 27-29 DEG C, and shaking speed is 150-170r/min, and the time is 6-9d.Preferably, it is described
The temperature of fermentation is 28 DEG C, and shaking speed is 160r/min, and the time is 7d.
Compared with prior art, beneficial effects of the present invention are：The present invention using liquid fermentation by the way of, by ferment
Condition is optimized, and improves the content of polysaccharide and flavones.The growth conditions of Phellinus 0.2% maltose be carbon source, 0.4% yeast
When extract is nitrogen source, polysaccharide and flavones content can obtain high value.
Description of the drawings
Fig. 1 is Phellinus polysaccharide, flavones content under the different carbon source percentage composition of embodiment 2；
Fig. 2 is Phellinus polysaccharide, the total output of flavones under the different carbon source percentage composition of embodiment 2；
Fig. 3 is Phellinus polysaccharide, flavones content under the different nitrogen sources percentage composition of embodiment 3；
Fig. 4 is Phellinus polysaccharide, flavones total output under the different nitrogen sources percentage composition of embodiment 3.
Specific embodiment
With reference to specific embodiment, the present invention is further elucidated, it should be understood that these embodiments are merely to illustrate the present invention
Rather than the scope of the present invention is limited, and after the present invention has been read, various equivalences of the those skilled in the art to the present invention
The modification of form falls within the application claims limited range.
Embodiment 1
The liquid fermentation method of Phellinus, including：
(1) actication of culture
Phellinus bacterial classification is seeded on PDA solid mediums, is placed in 28 DEG C of constant incubators and is cultivated 14d, for liquid
The inoculation of body culture.
(2) prepared by Phellinus one-level kind
The PDA liquid medium for having configured is dispensed in triangular flask, per bottled 100mL, 121 DEG C of high-pressure steam sterilizing pan
Use after sterilizing 30min.Aseptically, beaten on the solid medium for having activated with the aseptic card punch of a diameter of 6mm
Hole, selects 8 bacterial classification pieces and is put in PDA liquid medium, and triangular flask is put in 28 DEG C of shaking tables with 160r/min trainings after inoculation
Foster 7d, obtains Phellinus one-level kind.
(3) ferment
In superclean bench under aseptic condition, using seed beating crusher by Fermentative growth into first class inoculum smash, press
The inoculum concentration of 10% (v/v) is accessed in the triangular flask for the fluid nutrient medium of fermentation, and pH natures are placed in shaking table with 28
DEG C, the fermentation condition culture 7d of rotating speed 160r/min.
Measurement of the polysaccharide content method is as follows：
The preparation of polysaccharide：Phellinus bacterial classification is seeded to fluid nutrient medium, and fermented and cultured to mycelium pellet is filtered full of after culture medium,
Culture medium is filtered off with 60 mesh sieve, with distillation water washing, 60 DEG C of dryings on non-woven fabrics is placed on, dry mycelium is obtained, is pulverized,
Weigh.The mycelium of 0.05g is accurately weighed, in being soaked in the NaOH solution of 5mL 1mol/L, is extracted 1 hour in 100 DEG C of water-baths,
The extract of gained is polysaccharide crude.
The making of calibration curve：Weigh and be dried (60 degrees Celsius, 6 hours of thermostatic drying chamber) to the glucose 10mg of constant weight,
It is positioned over the volumetric flask of 100ml, plus deionized water constant volume, the as glucose standard of 0.1mg/ml.Precision draws glucose
Titer 0mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1mL, 1.2mL, 1.4mL, 1.6mL are added separately to each tool plug test tube
In, deionized water supplies every pipe 2ml, and each pipe adds 6% phenol of 1ml, shakes up rapidly.The concentrated sulfuric acid of 5ml is added, is put into
When should not add along tube wall, be directly added with beneficial to the concentrated sulfuric acid quickly and sample mix, stand 10 minutes, 100 DEG C of 15 points of water-baths
Clock.Room temperature is placed into after taking-up, each pipe absorbance is measured at 490nm wavelength.With the sugared concentration in test tube solution as abscissa,
The absorbance that each test tube is measured respectively is ordinate, makees calibration curve.Reagent blank is reference.
The assay of Phellinus polysaccharide：The content of Phellinus polysaccharide is determined using Phenol sulfuric acid procedure.Precision weighs Phellinus polysaccharide
0.02mL, deionized water complements to 2mL and is need testing solution, adds the making of amount of reagent and method of operating and calibration curve
It is identical, its absorbance is measured on spectrophotometer, according to calibration curve, calculate polyoses content.
The detection method of flavones content is as follows：
The extraction of flavones：The phellinus liteus 0.05g for pulverizing is weighed, 5mL70% ethanol is added, ultrasonic wave is placed on clear
Wash in instrument and extract 1h, obtain extract.
The assay of flavonoids from phellinus：Sample liquid 1mL is quantitatively pipetted in 25mL volumetric flasks, adds 30% ethanol to be supplemented to
10mL, adds 5%NaN020.2mL shakes up, and stands 5 minutes, adds 0.2mL 10%A1 (N03)3After standing 6 minutes, plus 2mL
4%NaOH, adds 70% ethanol to be supplemented to 50mL, and colorimetric estimation is carried out at 510nm wavelength, and reagent blank is reference.
The measure of rutin/ethanol calibration curve：Precision weighs 30mg rutins, and with 70% ethanol 50ml rutin standards are prepared
Liquid.Precision draws rutin titer 0ml, 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml and is added separately to each tool plug
In test tube, respectively add 5%NaNO2Solution 0.2mL, shakes up, and places 6min；Then respectively add the solution 0.2mL of 10%Al (NO3) 3, shake up
Place 6min；Then plus 4%NaOH solution 2mL, plus 70% ethanol is settled to 5mL, absorbance is surveyed at 510nm, does standard bent
Line.Calculate the flavones content of each sample.
The carbon source test of the different weight percentage of embodiment 2
Test method：In the step of embodiment 1 (3), carbon source (culture medium prescription of the fluid nutrient medium containing different weight percentage
For：Maltose, yeast extract 2g, peptone 2g, MgSO4·7H2O 0.5g, KH2PO44.6g, deionized water 1L), different hundred
In dividing the carbon source fluid nutrient medium of ratio, the content of maltose is respectively 0.5%, 1%, 2%, 4%, 6%.By the difference for having configured
The carbon source fluid nutrient medium of percentage is dispensed in triangular flask, every bottle of 100ml, is sealed with sealed membrane, then is tightened with rubber band.Often
Individual percentage does 3 repetitions, and sterilize 30min at 121 DEG C of high-pressure steam sterilizing pan.Other steps investigate carbon source pair with embodiment 1
The impact of Phellinus biologically active (polysaccharide and flavones).
Result of the test：
Carbon source is the most important nutrient content of fungi.It is the basic component of carbohydrate and protein, is again
Important energy source, in the fluid nutrient medium of the different weight percentage of identical carbon source, the percentage composition of maltose is respectively
0.5%th, 1%, 2%, 4%, 6%.By vaccinated culture medium with 28 DEG C, 160r/min culture 7d, mycelial polysaccharide is detected
Content and flavones content, as a result such as Fig. 1, table 1.
The significance of difference of Phellinus polysaccharide and flavones content under the different carbon source percentage composition of table 1
As shown in Fig. 1, table 1, Phellinus bacterial classification can grow in several culture mediums containing different carbon source percentage, but not
With impact of the carbon source to mycelia polysaccharide and flavones content, there were significant differences.When culture medium contains 2% maltose in Phellinus bacterial strain
Flavones and polyoses content it is higher, and the dense content of the polysaccharide when maltose content is 4% and 6% in mycelium is higher, but yellow
Ketone content is relatively low.While polysaccharide and flavones content is detected, the total output of polysaccharide and flavones is also carried out calculating, such as Fig. 2,
Table 2.
The significance of difference of Phellinus polysaccharide and flavones total output under the different carbon source percentage composition of table 2
As shown in Fig. 2, table 2, maltose content is at 2%, and the total output of polysaccharide and flavones is higher, in 4% and 6% content
Although lower polyoses content is high, low when flavones total output is compared with 2%, it is believed that, 2% carbon source content is chosen for culture Phellinus
Optimum carbon source percentage composition.
The nitrogen source test of the different weight percentage of embodiment 3
Test method：In the step of embodiment 1 (3), nitrogen source (culture medium prescription of the fluid nutrient medium containing different weight percentage
For：Maltose 20g, yeast extract, MgSO4·7H2O 0.5g, KH2PO44.6g, deionized water 1L), in different weight percentage
Nitrogen source fluid nutrient medium in, the percentage composition of yeast extract is respectively 0.1%, 0.2%, 0.4%, 0.8%, 1.2%.Will
The nitrogen source fluid nutrient medium of the different weight percentage for having configured is dispensed in triangular flask, every bottle of 100ml, is sealed with sealed membrane, then is used
Rubber band is tightened.Each percentage does 3 repetitions.121 DEG C of sterilizing 30min of high-pressure steam sterilizing pan.The same embodiment of other steps
1, investigate impact of the nitrogen source to Phellinus biologically active (polysaccharide and flavones).
Result of the test：
Nitrogen source is primarily referred to as in composition the material containing more nitrogen, mainly to things such as fungi synthetic protein, amino acid
Matter.Most of fungi can effectively utilize organic nitrogen and provide nutrition for itself.Growth of the nitrogen to Phellinus is benefited.In phase
In culture medium with the different percentage compositions of nitrogen source, the percentage composition of yeast extract is respectively 0.1%, 0.2%, 0.4%,
0.8%th, 1.2%.With the same terms culture Phellinus bacterial classification, the mycelium pellet to obtaining carries out polysaccharide and flavones content is determined, as a result
Such as Fig. 3, table 3.
The significance of difference of Phellinus polysaccharide and flavones content under the different nitrogen sources percentage composition of table 3
From Fig. 3, table 3 as can be seen that when yeast extract content is in 0.2% and 0.4%, polysaccharide and flavones content compared with
Height, than other nitrogen source contents, there were significant differences.In order to contrast the quality for drawing nitrogen concentration, to polysaccharide and the total output of flavones
It has been also carried out calculating, as a result such as Fig. 4, table 4.
The significance of difference of Phellinus polysaccharide and flavones total output under the different nitrogen sources percentage composition of table 4
Such as Fig. 4, shown in table 4, it can be seen that nitrogen source percentage composition is at 0.4%, and the total output of Phellinus polysaccharide and flavones is equal
Higher than other percentages, so, select the 0.4% most suitable nitrogen source fermented as Phellinus.
Cultivation temperature be 28 DEG C, shaking speed be 160r/min, fermentation time be 7d under conditions of, it is many with phellinus liteus
Sugar and flavones content are index, and research draws, 2% maltose is the most suitable percentage of carbon source, 0.4% yeast extract be nitrogen source most
The content of suitable percentage, polysaccharide and flavones can reach high value.
Claims (7)
1. it is a kind of improve Phellinus active component content method, it is characterised in that include：Phellinus seed liquor is inoculated in into liquid training
Fermented in foster base, containing carbon source and nitrogen source in described fluid nutrient medium, described carbon source is maltose, described nitrogen source
For yeast extract.
2. it is according to claim 1 improve Phellinus active component content method, it is characterised in that the maltose is in liquid
Concentration in body culture medium is 2-6%.
3. it is according to claim 1 improve Phellinus active component content method, it is characterised in that the yeast extract
Concentration in liquid medium within is 0.1-0.4%.
4. it is according to claim 1 improve Phellinus active component content method, it is characterised in that Liquid Culture during fermentation
Also contain MgSO in base4·7H2O 0.4-0.6 ‰, KH2PO4 4.5-5‰。
5. it is according to claim 1 improve Phellinus active component content method, it is characterised in that the Phellinus seed liquor
Preparation method be：PDA liquid medium will be inoculated in after Phellinus actication of culture, with 150-170r/min in 27-29 DEG C of shaking table
Culture 6-7d.
6. it is according to claim 1 improve Phellinus active component content method, it is characterised in that during fermentation, inoculum concentration
For 9-10%.
7. it is according to claim 1 improve Phellinus active component content method, it is characterised in that the temperature of the fermentation
For 27-29 DEG C, shaking speed is 150-170r/min, and the time is 6-9d.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611197331.4A CN106636215A (en) | 2016-12-22 | 2016-12-22 | Method of increasing active component content of phellinus igniarius |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611197331.4A CN106636215A (en) | 2016-12-22 | 2016-12-22 | Method of increasing active component content of phellinus igniarius |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106636215A true CN106636215A (en) | 2017-05-10 |
Family
ID=58834231
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611197331.4A Pending CN106636215A (en) | 2016-12-22 | 2016-12-22 | Method of increasing active component content of phellinus igniarius |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106636215A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110787194A (en) * | 2019-10-15 | 2020-02-14 | 郭红伟 | Phellinus igniarius paste and preparation method thereof |
| CN112812975A (en) * | 2021-01-19 | 2021-05-18 | 延边朝鲜族自治州农业科学院（延边特产研究所） | Culture medium for cultivating phellinus igniarius mother seeds and preparation method and cultivation method thereof |
-
2016
- 2016-12-22 CN CN201611197331.4A patent/CN106636215A/en active Pending
Non-Patent Citations (5)
| Title |
|---|
| ZHONGWEI LI等: "A Computational Method for Optimizing Experimental Environments for Phellinus igniarius via Genetic Algorithm and BP Neural Network", 《BIOMED RESEARCH INTERNATIONAL》 * |
| 刘凡等: "有利于桑黄产胞内黄酮的液体发酵培养基配方优化", 《蚕业科学》 * |
| 崔凤杰等: "基于响应曲面法优化桑黄菌深层发酵条件", 《食品科技》 * |
| 李朔: "桑黄液体发酵生产多糖的研究", 《中国优秀硕士学位论文全文数据库》 * |
| 祝明思等: "桑黄菌丝体液体发酵培养条件的研究", 《CHINA BREWING》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110787194A (en) * | 2019-10-15 | 2020-02-14 | 郭红伟 | Phellinus igniarius paste and preparation method thereof |
| CN112812975A (en) * | 2021-01-19 | 2021-05-18 | 延边朝鲜族自治州农业科学院（延边特产研究所） | Culture medium for cultivating phellinus igniarius mother seeds and preparation method and cultivation method thereof |
| CN112812975B (en) * | 2021-01-19 | 2023-11-17 | 延边朝鲜族自治州农业科学院（延边特产研究所） | Culture medium for culturing Phellinus linteus mother seeds, and preparation method and culturing method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105861346B (en) | One plant of high-yield urea, the abnormal Brunswick Durham yeast strain for producing wind taste and its application in food production | |
| CN105861348B (en) | The saccharomyces cerevisiae of one plant of high-yield urea and its application in food production | |
| CN105861345B (en) | One plant of high-yield urea, the pichia kudriavzevii for producing wind taste and its application in food fermentation | |
| CN110129207B (en) | Liquid fermentation medium for high-yield antioxidant cordyceps sobolifera mycelium and production method of antioxidant cordyceps sobolifera mycelium electuary | |
| CN102242154A (en) | Liquid fermentation method for producing paecilomyces cicadae mycelia and application of culture product | |
| CN101870740B (en) | Morchellaconica extracellular polysaccharide extractive and preparation method and application thereof | |
| CN101988036B (en) | Aureobasidium pullulans strain as well as preparation method and application thereof in producing pigment-free pullulan | |
| CN105695530B (en) | A kind of liquid fermentation medium of high yield Qaidam Agaricus bitorqui polysaccharide and application | |
| US20160230192A1 (en) | Triterpenoids high yielding strain of antrodia cinnamomea and use thereof | |
| CN105441340A (en) | Gibberellin GA4+7High-yield strain and application thereof | |
| CN110272835A (en) | One Accharomyces cerevisiae Saccharomyces cerevisiae ScEy01 and application | |
| CN110004066A (en) | A kind of mystery Ganoderma Varieties By Uv Induced and its artificial cultivation method and application | |
| CN106636215A (en) | Method of increasing active component content of phellinus igniarius | |
| CN104893978B (en) | One plant of haematococcus pluvialis ENN71 and its cultural method and application | |
| CN107119098A (en) | Addition growth factor produces bata-carotene and detection method during the fermentation | |
| CN102669320A (en) | Preparation method of sweet lucid ganoderma fermented tea and sweet lucid ganoderma fermented tea | |
| CN105766377B (en) | A kind of cultural method improving black fungus flavones content and type | |
| CN101914586A (en) | Method for preparing and purifying DON toxin | |
| CN107779337A (en) | A kind of extracted oil using olive produces the method that discarded object fermentation prepares fruit wine | |
| CN101589677A (en) | The training quality detection method of hypsizygus marmoreus produced by applying liquid strains | |
| CN103392512A (en) | Antrodia cinnamomea high yield triterpene bacterial strain and application thereof | |
| CN105166387B (en) | A kind of preparation method of ginkgo leaf biological feed additive | |
| CN114747423A (en) | Inonotus obliquus culture method and culture product thereof | |
| CN111607553B (en) | Ganoderma lucidum mycelium culture medium for high yield of polysaccharide and culture method thereof | |
| Gregori et al. | Growth characteristics and ergosterol content of Grifola frondosa in various solid-state substrates |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170510 |