CN108243834A - Antrodia camphorata fluid nutrient medium and preparation method thereof - Google Patents
Antrodia camphorata fluid nutrient medium and preparation method thereof Download PDFInfo
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- CN108243834A CN108243834A CN201810005885.2A CN201810005885A CN108243834A CN 108243834 A CN108243834 A CN 108243834A CN 201810005885 A CN201810005885 A CN 201810005885A CN 108243834 A CN108243834 A CN 108243834A
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Abstract
The present invention proposes fluid nutrient medium, which includes：Glucose, malt extract, peptone and monoterpenes compound, the monoterpenes compound are included selected from least one of geraniol, citronellol and camphor.Added at least one of monoterpenes compound geraniol, citronellol, camphor in the fluid nutrient medium, Antrodia camphorata triterpene substance is effectively facilitated and has been formed, and then significantly improved Antrodia camphorata total triterpene contents, and raw material is easy to get, it is at low cost, it is easy to operate, it is suitble to industrialized production.
Description
Technical field
The present invention relates to biotechnology, in particular it relates to fluid nutrient medium and preparation method thereof.
Background technology
Mushroom, red antrodia in antrodia (Antrodiacamphorata) also known as cinnamomum kanahirai hay mushroom, camphor tree cave, are subordinate to Basidiomycotina
(Basidiomycotina), Hymenomycetes (Hymenomycetes), Polyporaceae (Polyporaceae), antrodia karst
(Antrodia), it is a kind of rare medicinal fungus for originating in Taiwan.Antrodia camphorata is primary in TaiWan, China mountain area height above sea level 450~2000
The inner wall of the rotten heartwood of cinnamomum kanehirai (Cinnamomea kanehirai) trunk between rice more than distinctive age of tree a century, is platform
The authentic medicinal herbs in gulf are referred to as " Taiwan rarity ", " ruby in the forest of Taiwan ".
Antrodia camphorata contains abundant active material, it is now known that is more than 78 kinds of compounds, including triterpenes
(terpenoids), benzoic acid class (benzenoids), lignanoid (lignans), benzoquinones and its derivative
(benzoquinonederivatives), succinic acid and maleic acid and its derivative (uccinic and maleic
) and polysaccharide body etc. derivatives.Antrodia camphorata is in anti-inflammatory, liver protection, anti-liver cancer and anti-, anti-lung cancer, anti-breast cancer, anti-ovary in recent years
Cancer, inhibitor against colon carcinoma cells, anti-carcinoma of mouth, anti-oxidant, reduction nephritis of systemic lupus erythematousus and anti-platelet aggregation etc. are ground
Study carefully and all show that its future has high development potentiality.Antrodia camphorata is for after drinking or the dispelling effects of alcohol be still drank after a night also highly significant simultaneously.
One of the main active of triterpene substance for Antrodia camphorata, and the most especially, up to 200 kinds or more, be other
What mushroom can not be compared, there is anticancer, liver protection and other effects.Pharmacological research shows that triterpene has anticancer, anti-inflammatory, antiviral and liver protection
Etc. various actives.
Thus, the content for improving activated product triterpenes components in Cinnamomum kanahirai hay camphorata mycelium is to improve Antrodia camphorata drug value
Effective means.
Invention content
The application is that the discovery of following facts and problem and understanding are made based on inventor：
The Liquid Culture of the prior art can obtain a large amount of Cinnamomum kanahirai hay camphorata mycelium within a short period of time, make up Antrodia camphorata life
Long slowly this shortcoming, but the content of its activated product triterpenes components is but very low.Discovery based on the above problem, inventor
A kind of fluid nutrient medium is developed, in the fluid nutrient medium added with monoterpenes compound geraniol, citronellol, camphor at least
One of, it has effectively facilitated Antrodia camphorata triterpene substance and has been formed, and then significantly improved Antrodia camphorata total triterpene contents, and raw material is easy
, it is at low cost, it is easy to operate, it is suitble to industrialized production.
In the first aspect of the present invention, the present invention proposes a kind of fluid nutrient medium.According to an embodiment of the invention, it is described
Fluid nutrient medium includes：Glucose, malt extract, peptone and monoterpenes compound, the monoterpenes compound include
Selected from least one of geraniol, citronellol and camphor.Inventor has found, adds in monoterpenes compound in liquid medium
At least one of geraniol, citronellol and camphor can remarkably promote the formation of Antrodia camphorata triterpene substance.According to the present invention
The fluid nutrient medium of embodiment is remarkably improved total triterpene contents in Antrodia camphorata, and the raw material of culture medium for cultivating Antrodia camphorata
It is easy to get, it is at low cost, it is easy to operate, it is suitble to industrialized production.
According to an embodiment of the invention, aforesaid liquid culture medium can further include following additional technical feature at least
One of：
According to an embodiment of the invention, a concentration of the 0.05~0.1% of the geraniol.It should be noted that geraniol
A concentration of percent by volume.Inventor find, toward fluid nutrient medium in addition 0.05~0.1% geraniol, can significantly carry
The content of high Antrodia camphorata triterpene substance.Add in the unobvious very little, triterpene substance improves, add in it is too many, can promote it is non-must
The generation of by-product is needed, and causes to waste.
According to an embodiment of the invention, a concentration of the 0.05~0.1% of the citronellol.It should be noted that citronellol
A concentration of percent by volume.Inventor find, toward fluid nutrient medium in addition 0.05~0.1% geraniol, can significantly carry
The content of high Antrodia camphorata triterpene substance.Add in the unobvious very little, triterpene substance improves, add in it is too many, can promote it is non-must
The generation of by-product is needed, and causes to waste.
According to an embodiment of the invention, a concentration of the 0.01~0.05% of the camphor.It should be noted that camphor is dense
It spends for quality percentage by volume, unit is g/L.Inventor find, toward fluid nutrient medium in addition 0.05~0.1% spiceleaf
Alcohol can significantly improve the content of Antrodia camphorata triterpene substance.The unobvious very little, triterpene substance improves are added in, are added in too much,
It can promote the generation of nonessential by-product, and cause to waste.
According to an embodiment of the invention, a concentration of 20g/L of the glucose, a concentration of 20g/ of the malt extract
L, a concentration of 1g/L of the peptone.And then the yield of Antrodia camphorata further improves.
In the second aspect of the present invention, the present invention proposes a kind of method for preparing fluid nutrient medium noted earlier.According to
The embodiment of the present invention, the method includes：Water, glucose, malt extract and peptone are subjected to the first mixing and gone out
Bacterium is handled；Mixture after sterilizing and monoterpenes compound are subjected to the second mixed processing, to obtain the fluid nutrient medium,
Wherein, the monoterpenes compound is included selected from least one of geraniol, citronellol and camphor.Glucose, malt extraction
Object and peptone are the nutriments of culture medium, and sterile nutrient environment can be provided simultaneously for the growth of Antrodia camphorata after sterilization treatment
And monoterpenes compound is volatile at high temperature, and then by glucose, malt extract and peptone carry out first mixing and
The second mixed processing is carried out with monoterpenes compound again after sterilization treatment, the growth that may be either Antrodia camphorata provides sterile nutrition ring
Border, and can effectively avoid the loss of monoterpenes compound active ingredient.The liquid training obtained according to the method for the embodiment of the present invention
Base is supported, by adding at least one of monoterpenes compound geraniol, citronellol, camphor, significantly improves Antrodia camphorata total triterpene
Content, and raw material is easy to get, it is at low cost, it is easy to operate, it is suitble to industrialized production.
According to an embodiment of the invention, the above method can further include at least one following additional technical feature：
According to an embodiment of the invention, a concentration of the 0.05~0.1% of the geraniol.It should be noted that geraniol
A concentration of percent by volume.Inventor find, toward fluid nutrient medium in addition 0.05~0.1% geraniol, can significantly carry
The content of high Antrodia camphorata triterpene substance.Add in the unobvious very little, triterpene substance improves, add in it is too many, can promote it is non-must
The generation of by-product is needed, and causes to waste.
According to an embodiment of the invention, a concentration of the 0.05~0.1% of the citronellol.It should be noted that citronellol
A concentration of percent by volume.Inventor find, toward fluid nutrient medium in addition 0.05~0.1% geraniol, can significantly carry
The content of high Antrodia camphorata triterpene substance.Add in the unobvious very little, triterpene substance improves, add in it is too many, can promote it is non-must
The generation of by-product is needed, and causes to waste.
According to an embodiment of the invention, a concentration of the 0.01~0.05% of the camphor.It should be noted that camphor is dense
It spends for quality percentage by volume, unit is g/L.Inventor find, toward fluid nutrient medium in addition 0.05~0.1% spiceleaf
Alcohol can significantly improve the content of Antrodia camphorata triterpene substance.The unobvious very little, triterpene substance improves are added in, are added in too much,
It can promote the generation of nonessential by-product, and cause to waste.
According to an embodiment of the invention, a concentration of 20g/L of the glucose, a concentration of 20g/ of the malt extract
L, a concentration of 1g/L of the peptone.And then the yield of Antrodia camphorata further improves.
According to an embodiment of the invention, in first mixed processing, the water, glucose, malt sugar extract and
The amount ratio of peptone is 1000mL:20g:20g:1g.
According to an embodiment of the invention, the sterilization treatment is to carry out 30min under conditions of 121 degrees Celsius in temperature
's.
According to an embodiment of the invention, the monoterpenes compound include geraniol, citronellol and camphor, described second
In mixed processing, the amount ratio of mixture and the geraniol, citronellol and camphor after the sterilizing is 1000mL：0.5
~1mL:0.5-1mL:0.1-0.5g.
In the third aspect of the present invention, the present invention proposes a kind of cultural method of Antrodia camphorata.Implementation according to the present invention
Example, the cultural method include：Utilize foregoing fluid nutrient medium or the liquid prepared according to foregoing method
Body culture medium cultivates Antrodia camphorata.Cultural method according to embodiments of the present invention can promote Antrodia camphorata triterpene substance shape
Into, Antrodia camphorata total triterpene contents are significantly improved, and raw material is easy to get, it is at low cost, it is easy to operate, it is suitble to industrialized production.
According to an embodiment of the invention, the above method can further include at least one following additional technical feature：
According to an embodiment of the invention, Antrodia camphorata is cultivated using the culture medium, to obtain Antrodia camphorata mycelia
Body, the culture are that light culture 10-15d is carried out under conditions of temperature is 28 DEG C, rotating speed is 120rpm.In above-mentioned temperature, turn
Light culture 10-15d under conditions of speed, the Cinnamomum kanahirai hay camphorata mycelium total triterpene contents of acquisition further improve.
According to an embodiment of the invention, further comprise the Cinnamomum kanahirai hay camphorata mycelium being inoculated in solid culture matrix, with
Just Antrodia camphorata fructification is obtained, the solid culture matrix is soaked in the culture medium.Solid culture matrix is soaked in front
In the fluid nutrient medium, Cinnamomum kanahirai hay camphorata mycelium is inoculated in solid culture matrix later, the Antrodia camphorata fructification obtained
Middle total triterpene contents significantly improve.
According to an embodiment of the invention, before carrying out the culture, further comprise：Antrodia camphorata solid spawn is inoculated in
The culture medium.
According to a particular embodiment of the invention, it is described inoculation be by by size be 2*2cm Antrodia camphorata solid spawn block
It is inoculated in what the liquid culture medium was realized, inoculum concentration is：3 blocks of the culture medium inoculated Antrodia camphorata solid per 200mL
Strain block.Under above-mentioned inoculum density, be more suitable for the growth of Antrodia camphorata strain, further improve Antrodia camphorata yield and wherein effectively
The content of ingredient.
According to an embodiment of the invention, Antrodia camphorata is cultivated using the culture medium, it is real to obtain Antrodia camphorata
Body before carrying out the culture, further comprises：Antrodia camphorata solid spawn is inoculated in solid culture matrix, the solid training
Foster matrix is soaked in the culture medium.The content of triterpenoid in the Antrodia camphorata fructification obtained by the above method is notable
It improves.
According to an embodiment of the invention, Antrodia camphorata is cultivated using the culture medium, it is real to obtain Antrodia camphorata
Body before carrying out the culture, further comprises：Antrodia camphorata solid spawn is inoculated in the fluid nutrient medium, the inoculation
Be by by size be 5*5cm Antrodia camphorata solid spawn block be inoculated in what the liquid culture medium was realized, it is described cultivate be
Light culture is carried out under conditions of temperature is 28 DEG C, humidity is 80% 18 months.The Antrodia camphorata fructification obtained by the above method
In the content of triterpenoid significantly improve.
Specific embodiment
The embodiment of the present invention is described below in detail.The following examples are exemplary, it is intended to for explaining the present invention,
And it is not considered as limiting the invention.
General operation method：
1. ME fluid nutrient mediums are prepared, wherein glucose 20g/L, malt extract 20g/L, peptone 1g/L, shaking flask dress
It measures as 200mL/500mL, sterilize 30min in 121 DEG C.2. after culture medium cooling, add a concentration of 0.05-0.1%'s respectively
Geraniol, citronellol and a concentration of 0.01-0.05% camphors, and to be not added with as control.
3. being inoculated with Antrodia camphorata solid spawn to above-mentioned fluid nutrient medium, inoculum concentration is 3 pieces of the fungus block of every bottle of 2*2cm size,
Then in 28 DEG C of 120rpm light cultures 10-15d.
4. after the completion of culture, filtering bacterium solution obtains Antrodia camphorata bacterium ball, its triterpenes content is detected, processing group is 3.3-
4.6%, control group 1.5-1.9%.
Embodiment 1
1. ME fluid nutrient mediums are prepared, wherein glucose 20g/L, malt extract 20g/L, peptone 1g/L, shaking flask dress
It measures as 200mL/500mL, sterilize 30min in 121 DEG C.
2. after culture medium cooling, geraniol, citronellol and camphor are added respectively, make its final concentration be respectively
0.05%, 0.1%, 0.01%, and to be not added with as control.
3. taking Antrodia camphorata solid spawn, it is cut into the fungus block that size is 2*2cm, every bottle of 3 pieces of culture medium inoculated, then in 28 DEG C
120rpm light cultures 10d.
4. after the completion of culture, filtering bacterium solution obtains Antrodia camphorata bacterium ball, detects its total triterpene contents, adds the processing of geraniol
It is 3.8%, the processing for adding citronellol is 4.5%, and the processing for adding camphor is 3.7%, and it is 1.7% to compare.
Embodiment 2
1. ME fluid nutrient mediums are prepared, wherein glucose 20g/L, malt extract 20g/L, peptone 1g/L, shaking flask dress
It measures as 200mL/500mL, sterilize 30min in 121 DEG C.
2. after culture medium cooling, geraniol, citronellol and camphor are added respectively, it is respectively 0.1% to make its final concentration,
0.05%, 0.05%, and to be not added with as control.
3. taking Antrodia camphorata solid spawn, it is cut into the fungus block that size is 2*2cm, every bottle of 3 pieces of culture medium inoculated, then in 28 DEG C
120rpm light cultures 15d.
4. after the completion of culture, filtering bacterium solution obtains Antrodia camphorata bacterium ball, detects its total triterpene contents, adds the processing of geraniol
It is 4.0%, the processing for adding citronellol is 4.6%, and the processing for adding camphor is 3.3%, and it is 1.5% to compare.
Embodiment 3
1. ME fluid nutrient mediums are prepared, wherein glucose 20g/L, malt extract 20g/L, peptone 1g/L, shaking flask dress
It measures as 200mL/500mL, sterilize 30min in 121 DEG C.
2. after culture medium cooling, geraniol, citronellol and camphor are added respectively, make its final concentration be respectively
0.08%, 0.07%, 0.03%, and to be not added with as control.
3. taking Antrodia camphorata solid spawn, it is cut into the fungus block that size is 2*2cm, every bottle of 3 pieces of culture medium inoculated, then in 28 DEG C
120rpm light cultures 10d.
4. after the completion of culture, filtering bacterium solution obtains Antrodia camphorata bacterium ball, detects its total triterpene contents, adds the processing of geraniol
It is 4.3%, the processing for adding citronellol is 4.0%, and the processing for adding camphor is 3.6%, and it is 1.9% to compare.
Embodiment 4
1. ME fluid nutrient mediums are prepared, wherein glucose 20g/L, malt extract 20g/L, peptone 1g/L, shaking flask dress
It measures as 200mL/500mL, sterilize 30min in 121 DEG C.
2. after culture medium cooling, addition geraniol, citronellol and camphor, it is respectively 0.08% to make its final concentration,
0.07%, 0.03%, and to be not added with as control.
3. Cinnamomum kanahirai hay linden is packed, sterilize 4h in 121 DEG C, adds in aforesaid liquid culture medium to its bottom after cooling
Then 200mL is inoculated with the Antrodia camphorata solid spawn that size is 5*5cm, the light culture 18 under conditions of 28 DEG C of humidity 80% of temperature
A month to fruit-body formation.
4. collecting fructification, its total triterpene contents is detected, adds geraniol, citronellol, camphor processing is 6.5%, control
It is 4.6%.
By Examples 1 to 4 it is found that fluid nutrient medium according to embodiments of the present invention can significantly improve total three in Antrodia camphorata
Terpene content.
In the description of this specification, reference term " one embodiment ", " example ", " is specifically shown " some embodiments "
The description of example " or " some examples " etc. means specific features, structure, material or the spy for combining the embodiment or example description
Point is contained at least one embodiment of the present invention or example.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be in office
It is combined in an appropriate manner in one or more embodiments or example.In addition, without conflicting with each other, the skill of this field
Art personnel can tie the different embodiments or examples described in this specification and the feature of different embodiments or examples
It closes and combines.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is impossible to limitation of the present invention is interpreted as, those of ordinary skill in the art within the scope of the invention can be to above-mentioned
Embodiment is changed, changes, replacing and modification.
Claims (10)
1. a kind of fluid nutrient medium, which is characterized in that including：Glucose, malt extract, peptone and monoterpenes chemical combination
Object, the monoterpenes compound are included selected from least one of geraniol, citronellol and camphor.
2. fluid nutrient medium according to claim 1, which is characterized in that a concentration of the 0.05~0.1% of the geraniol；
Preferably, a concentration of the 0.05~0.1% of the citronellol；
Preferably, a concentration of the 0.01~0.05% of the camphor.
3. fluid nutrient medium according to claim 1, which is characterized in that a concentration of 20g/L of the glucose, the wheat
A concentration of 20g/L of bud extract, a concentration of 1g/L of the peptone.
A kind of 4. method for preparing any one of claims 1 to 3 fluid nutrient medium, which is characterized in that
Water, glucose, malt extract and peptone are subjected to the first mixing and sterilization treatment,
Mixture after sterilizing and monoterpenes compound are subjected to the second mixed processing, to obtain the fluid nutrient medium,
Wherein, the monoterpenes compound is included selected from least one of geraniol, citronellol and camphor.
5. according to the method described in claim 4, it is characterized in that, a concentration of the 0.05~0.1% of the geraniol；
Preferably, a concentration of the 0.05~0.1% of the citronellol；
Preferably, a concentration of the 0.01~0.05% of the camphor；
Optionally, a concentration of 20g/L of the glucose, a concentration of 20g/L of the malt extract, the peptone it is dense
It spends for 1g/L；
Optionally, in first mixed processing, the water, glucose, malt sugar extract and peptone amount ratio be
1000mL:20g:20g:1g；
Optionally, the sterilization treatment is to carry out 30min under conditions of 121 degrees Celsius in temperature；
Optionally, the monoterpenes compound includes geraniol, citronellol and camphor, described in second mixed processing
The amount ratio of mixture and the geraniol, citronellol and camphor after sterilizing is 1000mL：0.5~1mL:0.5-1mL:
0.1-0.5g。
6. a kind of cultural method of Antrodia camphorata, which is characterized in that utilize 3 any one of them liquid culture medium of claims 1 to 3
Or Antrodia camphorata is cultivated according to the fluid nutrient medium that claim 4~5 any one of them method prepares.
7. according to the method described in claim 6, it is characterized in that, cultivated using the culture medium Antrodia camphorata, so as to
Cinnamomum kanahirai hay camphorata mycelium is obtained, the culture is that light culture 10-15d is carried out under conditions of temperature is 28 DEG C, rotating speed is 120rpm；
Optionally, further comprise the Cinnamomum kanahirai hay camphorata mycelium being inoculated in solid culture matrix, it is real to obtain Antrodia camphorata
Body, the solid culture matrix are soaked in the culture medium.
8. the method according to the description of claim 7 is characterized in that before carrying out the culture, further comprise：By Antrodia camphorata
Solid spawn is inoculated in the culture medium.
9. according to the method described in claim 8, it is characterized in that, it is described inoculation be by by size be 2*2cm Antrodia camphorata
Solid spawn block is inoculated in what the liquid culture medium was realized, and inoculum concentration is：Described in 3 pieces of the culture medium inoculated per 200mL
Antrodia camphorata solid spawn block.
10. according to the method described in claim 6, it is characterized in that, cultivated using the culture medium Antrodia camphorata, so as to
Antrodia camphorata fructification is obtained, before carrying out the culture, is further comprised：Antrodia camphorata solid spawn is inoculated in solid medium
Body, the solid culture matrix are soaked in the culture medium；
Optionally, further comprise：Antrodia camphorata solid spawn is inoculated in the fluid nutrient medium, the inoculation is big by inciting somebody to action
The small Antrodia camphorata solid spawn block for 5*5cm is inoculated in what the liquid culture medium was realized, the culture be temperature be 28 DEG C,
Humidity carries out light culture 18 months under conditions of being 80%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110337991A (en) * | 2019-06-19 | 2019-10-18 | 三生源生物科技（天津）有限公司 | A kind of preparation and application of the inhibiting tumour cells agent based on Antrodia camphorata extract |
| US20210290709A1 (en) * | 2020-03-18 | 2021-09-23 | Greenyn Biotechnology Co., Ltd | Use of antrodia cinnamomea for increasing alcohol metabolism or/and hangover |
| US12138287B2 (en) | 2020-03-18 | 2024-11-12 | Greenyn Biotechnology Co., Ltd | Use of Antrodia cinnamomea for increasing alcohol metabolism or/and hangover |
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