CN117625421B - Composite microbial agent, dendrobium candidum fermentation liquor and preparation method and application thereof - Google Patents
Composite microbial agent, dendrobium candidum fermentation liquor and preparation method and application thereof Download PDFInfo
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Abstract
The invention relates to the field of microbial fermentation, and discloses a composite microbial agent, dendrobium candidum fermentation liquid, and a preparation method and application thereof. The composite microbial inoculum comprises saccharomyces cerevisiae and pichia pastoris, and the classification of the saccharomyces cerevisiae is named as:Saccharoymces cerevisiae 33the preservation number is CCTCC NO: M20232573, and the classification name of the Pichia pastoris is:Pichia pastorisGSthe preservation number is CCTCC NO: M20232572, and the CCTCC NO is preserved in China center for type culture Collection, address: the preservation date of the university of Wuhan, wuhan in China is 2023, 12 months and 15 days. The fermentation method comprises the step of fermenting dendrobium candidum liquid by adopting the two bacteria. The fermentation liquor obtained by the co-fermentation of the two bacteria has good effects of inhibiting the expression of NO and IL-6 and anti-inflammatory repair.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a composite microbial agent, dendrobium candidum fermentation liquid, and a preparation method and application thereof.
Background
Dendrobium officinale is a plant belonging to the genus Dendrobium of the family Orchidaceae, and is mainly distributed in the regions of Yunnan, guizhou, jiangxi and the like in China. Researches show that dendrobium candidum contains a plurality of active ingredients such as polysaccharide, polyphenol, flavone, dendrobine and the like, and can participate in regulating and controlling a plurality of physiological processes such as blood sugar, intestinal flora, oxidative stress, tumorigenesis and the like of a human body. The polysaccharide is the component with the highest content and the highest activity of the dendrobium candidum, has complex structure and large molecular weight, and has multiple effects of resisting oxidation, resisting tumor, relieving fatigue and the like. Common extraction methods of Dendrobium officinale active substances include soaking method, ultrasonic assisted method, enzymolysis method, etc. The microorganism contains abundant enzyme systems such as cellulase, amylase, pectase, protease and the like, and the fermentation can increase the availability of the active ingredients of the dendrobium candidum and improve the physiological activity of the dendrobium candidum through the action of the microorganism.
Under the industrial fermentation condition, microorganisms face a plurality of adversity stresses such as high temperature, high permeability, oxidation and the like, and the stresses inhibit the growth and the propagation of thalli to different degrees, so that the problems of low yield, high energy consumption and the like of biological fermentation are caused.
In view of this, the present invention has been made.
Disclosure of Invention
The invention aims to provide a composite microbial inoculum, dendrobium candidum fermentation liquor, and a preparation method and application thereof, and aims to solve at least one problem in the background art.
The invention is realized in the following way:
in a first aspect, the invention provides a composite microbial inoculant comprising saccharomyces cerevisiae and pichia pastoris, wherein the classification of the saccharomyces cerevisiae is named as follows:Saccharoymces cerevisiae 33the Chinese typical culture collection, address: the preservation date is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232573; pichia pastorisThe classification is named:Pichia pastorisGSthe Chinese typical culture collection, address: the preservation date of the Chinese university of Wuhan is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232572.
In an alternative embodiment, the ratio of CFU of Saccharomyces cerevisiae to Pichia pastoris is 1-3:1-3.
In a second aspect, the invention provides a preparation method of a dendrobium candidum fermentation liquid, which comprises the steps of inoculating Saccharomyces cerevisiae and Pichia pastoris into a dendrobium candidum fermentation medium for fermentation, wherein the dendrobium candidum fermentation medium contains dendrobium candidum and saccharide substances;
the classification of Saccharomyces cerevisiae is named:Saccharoymces cerevisiae 33the Chinese typical culture collection, address: the preservation date is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232573; the classification of pichia pastoris is named:Pichia pastorisGSthe Chinese typical culture collection, address: the preservation date is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232572;
optionally, the CFU ratio of Saccharomyces cerevisiae to Pichia pastoris is 1-3:1-3.
In an alternative embodiment, the dendrobium candidum fermentation medium comprises dendrobium candidum powder, sugar substances and deionized water; the dendrobium candidum fermentation medium comprises 50-75 g/L of dendrobium candidum powder, 9.5-10.5 g/L of sugar substances and pH of 6.0-7.5;
optionally, the carbohydrate is glucose.
In an alternative embodiment, the particle size of the dendrobium candidum powder is 60-200 meshes;
optionally, the dendrobium candidum fermentation medium is obtained by mixing the components and sterilizing.
In an alternative embodiment, the strain is concentrated to a concentration of 10 6 ~10 7 The concentration of the bacterial liquid and the bacterial strain of the saccharomyces cerevisiae with CFU/mL is 10 6 ~10 7 Inoculating CFU/mL of Pichia pastoris bacterial solution into the dendrobium candidum fermentation mediumFermenting;
alternatively, the strain was concentrated to 10 6 ~10 7 Inoculating CFU/mL mixed bacterial liquid containing Saccharomyces cerevisiae and Pichia pastoris into a dendrobium candidum fermentation medium for fermentation;
the volume ratio of the total inoculation amount of the bacterial liquid to the dendrobium candidum fermentation medium is 5% -10%.
In an alternative embodiment, after the fermentation is finished, sterilizing and filtering are sequentially carried out to obtain the dendrobium candidum fermentation liquid.
In an alternative embodiment, the filtration operation is to first filter the fermented medium with 200-300 mesh gauze to remove solid residues and then filter the filtrate with a 200 nm ceramic membrane.
In a third aspect, the invention provides a dendrobium candidum fermentation broth prepared by the preparation method according to any of the previous embodiments.
In a fourth aspect, the invention provides an application of the dendrobium candidum fermentation broth in preparation of medicines, skin care products or cosmetics with the effects of inhibiting the expression of NO and IL-6 or resisting inflammation and repairing.
The invention has the following beneficial effects:
the composite microbial inoculum provided by the invention contains the two yeasts, and the produced probiotics can increase the activity of the obtained fermentation broth by the synergistic effect of the two yeasts, so that the fermentation broth has good effects of inhibiting the expression of NO and IL-6 and resisting inflammation and repairing.
According to the fermentation method provided by the invention, as the specific two Saccharomyces cerevisiae and Pichia pastoris are adopted to ferment the dendrobium candidum together, the two bacteria are synergistic, and the produced probiotics can increase the activity of the obtained fermentation liquor, so that the fermentation liquor has good effects of inhibiting the expression of NO and IL-6 and resisting inflammation and repairing.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing growth of SLS medium during yeast screening;
FIG. 2 is a diagram showing protease hydrolysis circles during yeast screening;
FIG. 3 is a diagram showing cellulase hydrolysis circles during yeast screening;
FIG. 4 is a graph showing the effect of the comparative dendrobium candidum fermentation broth and the blank dendrobium candidum broth on NO secretion;
FIG. 5 is a graph showing the effect of the fermentation broth of Dendrobium officinale and the dendrobium officinale liquid of blank group on IL-6 secretion in each example and comparative example.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
The features and capabilities of the present invention are described in further detail below in connection with the examples.
The composite microbial inoculum provided by the embodiment of the invention comprises saccharomyces cerevisiae and pichia pastoris, and the classification of the saccharomyces cerevisiae is named as follows:Saccharoymces cerevisiae 33the Chinese typical culture collection, address: the preservation date is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232573; the classification of pichia pastoris is named:Pichia pastorisGSthe Chinese typical culture collection, address: the preservation date of the Chinese university of Wuhan is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232572.
The compound microbial inoculum contains the two yeasts, and the produced probiotics can increase the activity of the obtained fermentation broth through the synergistic effect of the two yeasts, so that the fermentation broth has good effects of inhibiting the expression of NO and IL-6 and resisting inflammation and repairing.
Preferably, the ratio of CFU of saccharomyces cerevisiae to pichia pastoris is 1 to 3:1 to 3 (e.g., 1:3, 1:2, 1:1, 2:1, or 3:1).
When the CFU ratio of the Saccharomyces cerevisiae to the Pichia pastoris in the composite microbial inoculum is 1-3:1-3, the composite microbial inoculum is used for fermenting the dendrobium candidum, so that the obtained fermentation liquid can inhibit the expression of NO and IL-6, and the anti-inflammatory repair effect is better.
In the embodiment, the cellulase and the protease are used as screening standards, and 2 Saccharomyces cerevisiae and Pichia pastoris strains capable of producing the cellulase and the protease are screened from the existing yeast library.
(1) SLS resistant Medium selection
Screening Medium (g/L): peptone 20.0, yeast extract 10.0, glucose 20.0, agar 20.0, and additional 1.0 and 2.0 mmol/L SLS (sodium lauryl sulfate) were added.
(2) Screening of cellulase-producing strains
Screening Medium (g/L): peptone 20.0, yeast extract 10.0, glucose 20.0, sodium carboxymethylcellulose 1.0, agar 20.0; sterilizing at 121deg.C under 0.1 Mpa for 20 min;
the incubation was carried out at 30℃for 3 days, after which the plates were immersed in 1.0% Congo red solution and incubated at room temperature for 30 min, and then decolorized with an appropriate amount of 1.0 mol/L sodium chloride solution 2 times for about 20 min each time, and then, whether or not transparent rings were present around the colonies was observed.
(3) Protease producing strain selection
Screening Medium (g/L): 10.0 parts of peptone, 5.0 parts of beef extract, 30.0 parts of skim milk powder, 5.0 parts of NaCl and 20.0 parts of agar; sterilizing at 121deg.C under 0.1 Mpa for 20 min;
culturing at 30deg.C for 3 days, and observing whether transparent rings are around the colony.
The test results are shown in Table 1 and FIGS. 1-4, and the two strains Y6 and Y8 obtained by screening are identified as Saccharomyces cerevisiae (Y6) and Saccharomyces pastorianus (Y8), respectively, with reference to commercial Pichia pastoris (CICC 1958) and certain SLS tolerance, protease and cellulase production capabilities.
TABLE 1 screening of strains
"+": the capacity is detected by the screening; "-": the capacity was not detected by this screening
The test results are shown in figures 1-4, and the two strains of Saccharomyces cerevisiae (left) and Pichia pastoris (right) obtained by screening have certain SLS tolerance, protease production and cellulase production.
The preparation method of the dendrobium candidum fermentation liquid provided by the embodiment of the invention comprises the steps of inoculating Saccharomyces cerevisiae and Pichia pastoris into a dendrobium candidum fermentation medium for fermentation, wherein the dendrobium candidum fermentation medium contains dendrobium candidum and saccharide substances;
the classification of Saccharomyces cerevisiae is named:Saccharoymces cerevisiae 33the Chinese typical culture collection, address: the preservation date is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232573; the classification of pichia pastoris is named:Pichia pastorisGSthe Chinese typical culture collection, address: the preservation date of the Chinese university of Wuhan is 2023, 12 months and 15 days, and the preservation number is CCTCC NO: M20232572.
According to the preparation method of the dendrobium candidum fermentation liquid, the dendrobium candidum is fermented by adopting the two bacteria, so that the two bacteria can play a synergistic role, and the obtained fermentation liquid has a good effect of inhibiting the expression of NO and IL-6 and resisting inflammation and repairing.
Specifically, the preparation method comprises the following steps:
s1, preparing bacterial liquid
The Saccharomyces cerevisiae and Pichia pastoris used in the application are screened out by the screening method provided by the above.
The two yeasts thus selected were inoculated into YPD medium (seed medium) Culturing at 30 ℃ and 200 rpm for 12-24 hours until the concentration of the bacterial cells reaches 10 6 ~10 7 Obtaining bacterial liquid in CFU/mL;
YPD Medium (g/L) above: peptone 20.0, yeast extract 10.0, glucose 20.0; sterilizing at 121deg.C under 0.1 Mpa for 20 min.
S2, preparing dendrobium candidum fermentation medium
Cutting off dendrobium candidum, crushing by a crusher, and sieving by a 60-200 mesh sieve to obtain dendrobium candidum powder;
adding dendrobium candidum powder into deionized water according to the addition amount of 50-75 g/L (50 g/L, 60g/L, 70 g/L or 75 g/L), adding sugar substances according to the addition amount of 9.5-10.5 g/L (9.5 g/L, 10g/L or 10.5 g/L), regulating the pH of the system to 6.0-7.5, and sterilizing to obtain the fermentation medium.
Optionally, the carbohydrate is glucose.
Optionally, the sterilization condition is 121 ℃ and the sterilization is carried out for 20 min under the condition of 0.1 Mpa.
S3, inoculating and fermenting
Mixing the two bacterial solutions in a ratio of 1:3-3:1 to obtain a composite bacterial agent, and inoculating the composite bacterial agent into a dendrobium candidum fermentation medium;
or inoculating the two bacterial liquids obtained in the step S1 into a dendrobium candidum fermentation medium, wherein the volume ratio of the saccharomyces cerevisiae bacterial liquid to the pichia pastoris bacterial liquid is 1:3-3:1.
The volume ratio of the total inoculation amount of the bacterial liquid to the dendrobium candidum fermentation medium is 5% -10% (for example, 5%, 8% or 10%), and the fermentation is stopped when the residual sugar is 0 after fermentation culture at 30+/-2 ℃;
after fermentation, sterilizing and filtering in sequence to obtain dendrobium candidum fermentation liquor;
optionally, the sterilization condition is 121 ℃ and the sterilization is carried out for 20 min under the condition of 0.1 Mpa.
Alternatively, the filtration operation is to first filter the fermented medium with 200-300 mesh gauze to remove solid residues, and then filter the filtrate with a 200 nm ceramic membrane.
The dendrobium candidum fermentation liquor provided by the embodiment of the invention is prepared by adopting the preparation method provided by the embodiment of the invention.
The embodiment of the invention also provides application of the dendrobium candidum fermentation liquor in preparing medicines, skin care products or cosmetics with the effects of inhibiting the expression of NO and IL-6 or resisting inflammation and repairing.
The present invention will be specifically described with reference to a plurality of examples and comparative examples, wherein the same bacterial solutions used in the examples and comparative examples are produced by the same batch of culture and the number of strains contained therein is substantially the same.
Example 1
Concentration is set to 10 6 ~10 7 CFU/mL Saccharomyces cerevisiae bacteria liquid with concentration of 10 6 ~10 7 The CFU/mL Pichia pastoris bacterial liquid is inoculated into the dendrobium candidum fermentation medium for fermentation according to the volume ratio of 1:1, and the ratio of the total bacterial liquid volume of inoculation to the volume of the dendrobium candidum fermentation medium is 5%.
The dendrobium candidum fermentation medium contains 60g/L dendrobium candidum powder, 10g/L glucose and 6.5-7.5 pH.
Example 2
Concentration is set to 10 6 ~10 7 CFU/mL Saccharomyces cerevisiae bacteria liquid with concentration of 10 6 ~10 7 The CFU/mL Pichia pastoris bacterial liquid is inoculated into the dendrobium candidum fermentation medium for fermentation according to the volume ratio of 3:1, and the ratio of the total bacterial liquid volume of inoculation to the volume of the dendrobium candidum fermentation medium is 5%.
The dendrobium candidum fermentation medium contains 50g/L of dendrobium candidum powder, 10g/L of glucose and 6.0-7.0 of pH.
Example 3
Concentration is set to 10 6 ~10 7 CFU/mL Saccharomyces cerevisiae bacteria liquid with concentration of 10 6 ~10 7 The CFU/mL Pichia pastoris bacterial liquid is inoculated into the dendrobium candidum fermentation medium for fermentation according to the volume ratio of 1:3, and the ratio of the total bacterial liquid volume of inoculation to the volume of the dendrobium candidum fermentation medium is 5%.
The dendrobium candidum fermentation medium contains 75g/L dendrobium candidum powder, 10g/L glucose and 6.0-7.0 pH.
Comparative example 1
This comparative example is substantially the same as example 1, except that: equal amount of saccharomyces cerevisiae bacteria liquid is used for replacing pichia pastoris bacteria liquid.
Comparative example 2
This comparative example is substantially the same as example 1, except that: equal amount of Pichia pastoris liquid is used to replace Saccharomyces cerevisiae liquid.
Comparative example 3
This comparative example is substantially the same as example 1, except that: the fermentation was inoculated with a conventional common yeast equivalent to example 1, which is a commercially available Saccharomyces cerevisiae (CICC 1210).
Experimental example
Blank DO: adding deionized water into dendrobium candidum powder, performing ultrasonic treatment for 30 min, and centrifuging at 4000 rpm for 10 min to obtain unfermented dendrobium candidum extract;
experimental group FDO: fermentation broths prepared in examples 1 to 3 and comparative examples 1 to 3;
the performance of the blank and individual experimental group samples was tested. The method comprises the following steps:
(1) Secretion of NO
NO acts as an important inflammatory signaling molecule, and its secreted amount can reflect the intensity of inflammation. Macrophage RAW264.7 was seeded into 6-well plates at a density of 1X 106/mL at 5% CO 2 24 h were cultured in a biochemical incubator at 37 ℃. After the cell fusion degree reaches 80-90%, DO and FDO prepared by different concentrations of DMEM are added into each hole of the sample group for continuous culture 8 h for pretreatment, and a blank group and a negative control group are added into the DMEM for culture. The sample group and the negative control group 6 h were stimulated with 1. Mu.g/mL LPS. After the stimulation, the cell culture broth was collected and centrifuged at 12,000 rpm for 5 min, and the supernatant was collected. The amount of NO in the solution was measured by Griess reagent method, 50. Mu.L of supernatant was added to 96-well plates, 50. Mu.L of Griess reagent A (1% sulfanilamide, 2% phosphoric acid aqueous solution) was added to each well, absorbance was measured at 550 nm, 50. Mu.L of Griess reagent B (0.1% N- (1-naphthyl) ethylenediamine hydrochloride solution) was added to each well, and after reaction at room temperature for 10 minutes in the absence of light, absorbance was measured at 550 nm. By NaNO 2 The standard curve was determined after preparing the gradient standard solution.
(2) Secretion of IL-6
Interleukin-6 (IL-6) is a multifunctional inflammatory cytokine that plays an important role in the inflammatory response. The cells were cultured as described above to collect cell supernatants, and inflammatory factors (IL-6) were detected using ELISA kits.
The test results are shown in figures 4 and 5, the NO and inflammatory factor IL-6 secretion of RAW264.7 cells is obviously increased under the stimulation of 1 mug/mL LPS, the secretion level of NO and IL-6 in macrophages induced by LPS is obviously reduced along with the increase of the administration concentration of DO and FDO, and the FDO group can better reduce the secretion of NO and IL-6, which indicates that the anti-inflammatory effect of dendrobium candidum is improved by fermentation.
As can be seen from the test results shown in fig. 4 and 5, in the FDO group, examples 1 to 3 are significantly better in anti-inflammatory effect than comparative examples 1 to 3; comparing example 1 with comparative examples 1 and 2, the results show that fermentation liquor obtained by simultaneous inoculation and fermentation of Saccharomyces cerevisiae and Pichia pastoris has better anti-inflammatory effect, and the two yeasts have synergistic effect in improving anti-inflammatory effect by fermentation; comparing example 1 with comparative example 3, the anti-inflammatory effect of the fermentation broth of example 1 is better, demonstrating that the composite microbial agent fermentation of the invention can improve the anti-inflammatory property of the fermentation broth compared with the existing yeast fermentation.
In conclusion, the compound microbial inoculum provided by the embodiment of the invention contains the two yeasts, and the produced probiotics can increase the activity of the obtained fermentation broth through the synergistic effect of the two yeasts, so that the fermentation broth has good effects of inhibiting the expression of NO and IL-6 and resisting inflammation and repairing.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A composite microbial agent is characterized by comprisingSaccharomyces cerevisiaeSaccharoymces cerevisiae) Pichia pastorisPichia pastoris) The saccharomyces cerevisiae is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M20232573; the Pichia pastoris is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M20232572;
the CFU ratio of the Saccharomyces cerevisiae to the Pichia pastoris is 1-3:1-3.
2. The method for preparing the dendrobium candidum fermentation liquid is characterized by comprising the step of inoculating the composite microbial inoculum according to claim 1 into a dendrobium candidum fermentation medium for fermentation, wherein the dendrobium candidum fermentation medium contains dendrobium candidum and saccharide substances.
3. The preparation method according to claim 2, further comprising at least one of the following features (1) - (3):
(1) The dendrobium candidum fermentation medium comprises the following components of dendrobium candidum powder, sugar substances and deionized water; the dendrobium candidum powder comprises 50-75 g/L of sugar, 9.5-10.5 g/L of sugar and 6.0-7.5 of pH of a dendrobium candidum fermentation medium;
(2) The saccharide is glucose;
(3) The dendrobium candidum fermentation medium is obtained by mixing the components and sterilizing.
4. The preparation method of claim 3, wherein the particle size of the dendrobium candidum powder is 60-200 meshes.
5. The method according to claim 2, wherein the strain concentration is 10 6 ~10 7 CFU/mL of the saccharomyces cerevisiae bacterial liquid and strain concentration is 10 6 ~10 7 Inoculating CFU/mL of the Pichia pastoris bacterial solution into the dendrobium candidum fermentation medium for fermentation;
alternatively, the strain is subjected toThe concentration is 10 6 ~10 7 Inoculating CFU/mL mixed bacterial liquid containing the saccharomyces cerevisiae and the pichia pastoris into the dendrobium candidum fermentation medium for fermentation;
the volume ratio of the total inoculation amount of the bacterial liquid to the dendrobium candidum fermentation medium is 5% -10%.
6. The preparation method according to claim 2, wherein after the fermentation is completed, the dendrobium candidum fermentation liquid is obtained by sequentially performing sterilization and filtration operations.
7. The method according to claim 6, wherein the filtration is performed by first removing solid residues from the fermented medium with a 200-300 mesh gauze and then filtering the filtrate with a 200 nm ceramic membrane.
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| CN113966832A (en) * | 2020-07-22 | 2022-01-25 | 安琪纽特股份有限公司 | Dendrobium nobile fermentation product and preparation method and application thereof |
| CN114831918A (en) * | 2022-05-23 | 2022-08-02 | 合肥卡迪尔生物科技有限公司 | Traditional Chinese medicine composition fermentation liquor, preparation method and application thereof |
| CN115572738A (en) * | 2022-11-02 | 2023-01-06 | 广州悦荟化妆品有限公司 | Dendrobium officinale fermentation liquor with anti-inflammatory, relieving and repairing effects as well as preparation method and application thereof |
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| CN113966832A (en) * | 2020-07-22 | 2022-01-25 | 安琪纽特股份有限公司 | Dendrobium nobile fermentation product and preparation method and application thereof |
| CN114831918A (en) * | 2022-05-23 | 2022-08-02 | 合肥卡迪尔生物科技有限公司 | Traditional Chinese medicine composition fermentation liquor, preparation method and application thereof |
| CN115572738A (en) * | 2022-11-02 | 2023-01-06 | 广州悦荟化妆品有限公司 | Dendrobium officinale fermentation liquor with anti-inflammatory, relieving and repairing effects as well as preparation method and application thereof |
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