CN114699343B - Phellinus linteus fermentation product and preparation method thereof - Google Patents
Phellinus linteus fermentation product and preparation method thereof Download PDFInfo
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- CN114699343B CN114699343B CN202210049751.7A CN202210049751A CN114699343B CN 114699343 B CN114699343 B CN 114699343B CN 202210049751 A CN202210049751 A CN 202210049751A CN 114699343 B CN114699343 B CN 114699343B
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- 230000004151 fermentation Effects 0.000 title claims abstract description 85
- 241000001727 Tropicoporus linteus Species 0.000 title claims abstract description 26
- 238000004519 manufacturing process Methods 0.000 title claims description 10
- 238000002360 preparation method Methods 0.000 title abstract description 13
- 235000002595 Solanum tuberosum Nutrition 0.000 claims abstract description 42
- 244000061456 Solanum tuberosum Species 0.000 claims abstract description 42
- 241000123330 Fomes fomentarius Species 0.000 claims abstract description 39
- 239000000047 product Substances 0.000 claims abstract description 30
- 239000000758 substrate Substances 0.000 claims abstract description 25
- 239000000843 powder Substances 0.000 claims abstract description 17
- 238000000034 method Methods 0.000 claims abstract description 16
- 239000007788 liquid Substances 0.000 claims abstract description 12
- 230000001954 sterilising effect Effects 0.000 claims abstract description 5
- 230000004913 activation Effects 0.000 claims abstract description 4
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
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- PGNRLPTYNKQQDY-UHFFFAOYSA-N 2,3-dihydroxyindole Chemical compound C1=CC=C2C(O)=C(O)NC2=C1 PGNRLPTYNKQQDY-UHFFFAOYSA-N 0.000 description 1
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- 208000003351 Melanosis Diseases 0.000 description 1
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- DGLRDKLJZLEJCY-UHFFFAOYSA-L disodium hydrogenphosphate dodecahydrate Chemical compound O.O.O.O.O.O.O.O.O.O.O.O.[Na+].[Na+].OP([O-])([O-])=O DGLRDKLJZLEJCY-UHFFFAOYSA-L 0.000 description 1
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- 239000002023 wood Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Birds (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Microbiology (AREA)
- Pain & Pain Management (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Rheumatology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present disclosure provides a method of preparing a fomes fomentarius ferment, comprising: step one, carrying out activation, purification and expansion culture treatment on phellinus linteus to obtain seed liquid; step two, inoculating the seed solution to a fermentation substrate for fermentation culture treatment to obtain fermentation liquor; the fermentation substrate is potato homogenate; and thirdly, separating and sterilizing the fermentation liquor to obtain supernatant. Also disclosed are fomes fomentarius fermented products, including fomes fomentarius fermented solutions and dry powders thereof, made therefrom. According to the preparation method of the fomes fomentarius fermentation product, fomes fomentarius is inoculated into potato homogenate for fermentation, and the obtained fermentation product has good skin anti-inflammatory protection and restoration effects and can be used as an effective component to be added into a cosmetic formula.
Description
Technical Field
The present disclosure belongs to the technical field of biological fermentation, and in particular relates to a fomes fomentarius ferment and a preparation method thereof.
Background
Phellinus linteus (La Ding Ming is Fomes fomentarius), also called Phellinus linteus belonging to Basidiomycetes, polyporales, polyporaceae, phellinus genus, which is white rot fungus, is usually parasitic on broad-leaved trees such as birch, poplar, pear, apple, etc., and its fruiting body is medicinal part, and can be used for resolving food stagnation and removing blood stasis. Phellinus linteus is fungus with important application value, and the fruiting body and mycelium of the fungus contain various bioactive substances, and the representative substances are wood hoof polysaccharide, which has the effects of resisting oxidation, promoting urination, reducing fever, relieving pain, diminishing inflammation, resisting tumor and the like.
However, reports on the cosmetic application of fomes fomentarius are currently blank. The inventor finds that the ferment obtained by the synergistic fermentation of the fomes fomentarius and the pearl has excellent skin anti-inflammatory and repairing effects and whitening effects, and can be used as an effective component for cosmetics.
Disclosure of Invention
The following presents a simplified summary of the disclosure in order to provide a basic understanding of some aspects of the disclosure. It should be understood that this summary is not an exhaustive overview of the disclosure. It is not intended to identify key or critical elements of the disclosure or to delineate the scope of the disclosure. Its purpose is to present some concepts in a simplified form as a prelude to the more detailed description that is discussed later.
In order to solve the technical problems, the technical scheme provided by the present disclosure is as follows:
in a first aspect, the present disclosure provides a method of preparing a fomes fomentarius ferment comprising:
step one, carrying out activation, purification and expansion culture treatment on phellinus linteus to obtain seed liquid;
step two, inoculating the seed solution to a fermentation substrate for fermentation culture treatment to obtain fermentation liquor; the fermentation substrate is potato homogenate;
and thirdly, separating and sterilizing the fermentation liquor to obtain supernatant.
Further, in the preparation method of the fomes fomentarius fermented product, pearl powder is also added into the fermented substrate; the pearl powder is 1-10wt% (such as 2wt%, 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt%, etc.) of the potato homogenate; more preferably 1-5wt% (e.g., 1.5wt%, 2wt%, 3wt%, 4wt%, 4.5wt%, etc.).
Further, in the preparation method of the fomes fomentarius fermentation product, the potato homogenate is prepared by homogenizing fresh potato slices and water by a homogenizer, and dissolving, wherein the content of the potato is 1-5wt% (such as 1.2wt%, 1.5wt%, 2.0wt%, 2.5wt%, 3wt%, 3.5wt%, 4wt%, 4.5wt%, and the like).
Further, in the preparation method of the fomes fomentarius fermentation product, the potato homogenate also comprises 0.3wt% of monopotassium phosphate, 0.15wt% of magnesium sulfate, 0.05wt% of glucose and 0.2wt% of soybean peptide.
The fermentation substrate is subjected to a conventional sterilization treatment, such as high temperature sterilization at 115 ℃ for 20min, prior to inoculation.
Further, in the preparation method of the fomes fomentarius fermentation product, the mycelium volume percentage of the seed liquid is more than 80% (such as 82%, 85%, 90%, etc.); the inoculation proportion of the Phellinus baumii, namely the volume ratio of the bacterial liquid to the pearl liquid fermentation medium (fermentation substrate) is 1% -10% (such as 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, etc.).
Further, the fomes fomentarius is purchased from China general microbiological culture Collection center and is selected from at least one of the following: CGMCC5.1494, CGMCC 5.983, GCMCC5.96 or GCMCC5.913.
Further, in the above method for producing a fomes fomentarius fermented product, the fermentation culture treatment is carried out at a temperature of 25 to 35 ℃ (e.g., 26 ℃,28 ℃, 30 ℃, 32 ℃, 34 ℃ and the like) for a time of 48 to 96 hours (e.g., 52 hours, 60 hours, 72 hours, 84 hours, 88 hours, 92 hours and the like).
Further, the fermentation culture treatment is performed in a shaking table at a rotation speed of 150r/min-200r/min (such as 155r/min, 160r/min, 165r/min, 170r/min, 180r/min, 190r/min, 195r/min, etc.).
Further, in the preparation method of the fomes fomentarius fermentation product, the separation treatment adopts a centrifugal method; more preferably, the centrifugal speed is 4500r/min-5500r/min (such as 4600r/min, 4800r/min, 5000r/min, 5200r/min, 5400r/min, etc.), and the centrifugal time is 10min-20min (such as 12min, 15min, 18min, etc.).
In a second aspect, the present disclosure provides a fomes fomentarius ferment produced by the above-described production process. The Phellinus linteus pearl fermentation product comprises Phellinus linteus fermentation liquid and dry powder thereof, wherein the dry powder can be prepared by freeze drying method or spray drying method.
In a third aspect, the present disclosure also provides a cosmetic comprising the above phellinus linteus ferment. The cosmetic can be facial mask, essence, cream, lotion, etc.
Compared to the prior art, the benefits of the present disclosure include, but are not limited to:
1. according to the preparation method of the fomes fomentarius fermentation product, fomes fomentarius is inoculated into potato homogenate for fermentation, and the obtained fermentation product has good skin anti-inflammatory protection and restoration effects;
2. according to the preparation method of the fomes fomentarius fermentation product, fomes fomentarius and pearl are subjected to synergistic fermentation to prepare the fermentation product with excellent anti-inflammatory performance and whitening performance, and the fermentation product can be used as an effective component to be added into a cosmetic formula to prepare cosmetics such as facial masks, essence, sun cream, emulsion and the like;
3. the fomes fomentarius fermentation product provided by the disclosure has the characteristics of strong efficacy, low cost, simplicity in operation, green safety and the like as an efficacy component of cosmetics, and has stronger practicability and popularization value.
Drawings
FIG. 1 is a schematic diagram showing the detection results of the anti-inflammatory effect of the protection treatment of the phellinus linteus pearl fermentate in example 2, and the indexes are as follows: IL-1 beta;
FIG. 2 is a schematic diagram showing the detection results of the anti-inflammatory effect of the protection treatment of the extract of the pearl potato in example 2, which is an index: IL-1 beta;
FIG. 3 is a schematic diagram showing the detection results of the anti-inflammatory effect of the protection treatment of the phellinus linteus pearl ferments with different concentrations in example 2, and the indexes are as follows: IL-1 beta;
FIG. 4 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the Phellinus linteus pearl fermentate in example 2, which is an index: IL-1 beta;
FIG. 5 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the extract of the potato in example 2, the indexes: IL-1 beta;
FIG. 6 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the phellinus linteus pearl ferments with different concentrations in example 2, and the indexes are as follows: IL-1 beta;
FIG. 7 is a schematic diagram showing the detection results of the anti-inflammatory effect of the repair treatment of the extract of the potato in example 2, the index: IL-8;
FIG. 8 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the phellinus linteus pearl ferments with different concentrations in example 2, and the indexes are as follows: IL-8.
Detailed Description
The following examples further illustrate the invention but are not to be construed as limiting the invention. Modifications and substitutions of the methods, steps or conditions of the present invention without departing from the spirit and nature of the invention are intended to be within the scope of the present invention.
Technical aspects of the present disclosure will be described hereinafter with reference to exemplary embodiments. The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The fomes fomentarius used in the examples are commercial products purchased from the China general microbiological culture Collection center with a accession number: CGMCC5.1494.
EXAMPLE 1 preparation of Phellinus linteus fermentate
(1) Activating strains: bacterial colonies are picked from the inclined plane of preserved fomes fomentarius CGMCC5.1494 (laboratory number 5372) and placed in a glucose potato agar culture medium for 7d of culture and activation at 28 ℃, then the obtained single bacterial colonies are inoculated into 100mL of glucose potato liquid culture medium for 7d of culture at 28 ℃ and 180rpm, so that fomes fomentarius Kong Junchong seed liquid is obtained, and the volume ratio of mycelia accounts for 80% of the whole seed liquid.
(2) Preparing a fermentation substrate: according to the settings of Table 1 below, a certain mass ratio of pearl powder (1-10 wt%, such as 1wt%, 5wt%, 10 wt%) was added to potato homogenate, wherein the potato homogenate was prepared by adding 20g of potato slices to 1000ml of mineral water (containing 0.3% potassium dihydrogen phosphate, 0.15% magnesium sulfate, 0.05% glucose, 0.2% soybean peptide), adding to a homogenizer, homogenizing, dissolving, and packaging. The fermentation substrate is sterilized prior to inoculation.
(3) Inoculating and shaking: according to the inoculation amount of 5% by volume, 5mL of seed solution is inoculated into a 250mL triangular flask filled with 100mL of fermentation substrate, and fermented in a shaking table at a rotation speed of 180r/min and a culture temperature of 28 ℃ for 48h.
(4) And (3) centrifuging: the fermentation broth was centrifuged at 4800r/min for 10min, the supernatant was taken, sterilized, and the obtained sample numbers were as shown in Table 1.
The example also provides a control group with a simple fermentation substrate of 2% potato homogenate as described above, with 0.3% potassium dihydrogen phosphate, 0.15% magnesium sulfate, 0.05% glucose, 0.2% soy peptide. See table 1 for details.
TABLE 1 fermentation regime and experimental group designation for example 1
Experiment group number | Fermentation substrate | Inoculation situation |
TD-0 | Simple fermentation of substrates | Not inoculated with |
TD-5372 | Simple fermentation of substrates | Inoculating 5372 strain |
1%ZZ-TD-5372 | Contains 1% of pearlFermentation substrate for flour | Inoculating 5372 strain |
5%ZZ-TD-5372 | Fermentation substrate containing 5% pearl powder | Inoculating 5372 strain |
10%ZZ-TD-5372 | Fermentation substrate containing 10% pearl powder | Inoculating 5372 strain |
1%ZZ-TD | Fermentation substrate containing 1% pearl powder | Not inoculated with |
5%ZZ-TD | Fermentation substrate containing 5% pearl powder | Not inoculated with |
10%ZZ-TD | Fermentation substrate containing 10% pearl powder | Not inoculated with |
The main components of the product obtained in example 1, including protein, amino acid and calcium content, were tested and the results are shown in tables 2-4 below.
TABLE 2 amino acid content (umol/mL) of stock solution or fermentation stock solution samples
TABLE 3 protein content of stock solutions or fermentation stock solutions sample (ug/mL)
TABLE 4 calcium content of stock solutions or fermentation stock solutions
Experiment group number | Calcium (mg/kg) | Experiment group number | Calcium (mg/kg) |
TD-0 | 12.4 | TD-5372 | 1.3 |
1%ZZ-TD | 39.0 | 1%ZZ-TD-5372 | 179.5 |
5%ZZ-TD | 33.1 | 5%ZZ-TD-5372 | 167.7 |
10%ZZ-TD | 28.2 | 10%ZZ-TD-5372 | 25.3 |
EXAMPLE 2 analysis of efficacy of Phellinus linteus ferments
1. Toxicity and protective action of Phellinus linteus fermentation on human skin fibroblasts (HSF cells)
1. Establishment of oxidative stress model
HSF cells were 1X 10 per well 4 Inoculating into 96-well plate, culturing overnight in cell culture box, discarding culture medium, and adding 100 μl of different concentrations of H 2 O 2 (50~1000μmol·L -1 ) 5 replicates for each group and no H was added to the control group 2 O 2 . After 1, 2, 3, 4h of stimulation, the OD of each well was measured at 490nm using the MTT method. The HSF cell viability was obtained (see fig. 1), and hydrogen peroxide concentration at which the cell viability was 50% was selected as the modeling concentration. In this example, 100. Mu. Mol.L -1 H of (2) 2 O 2 HSF cells were treated for 2h modeling and cell viability was (49.74.+ -. 2.99)%.
2. Sample pair H 2 O 2 Protection and repair detection of induced oxidative stress model
HSF cells were 1X 10 per well 4 The samples were inoculated into 96-well plates and cultured overnight in a cell culture incubator, and a Control group (Control), a Model group (Model), and each sample group (stock solution or fermentation stock solution in example 1) were each set with 3 replicates.
The repair effect detection means that after cell injury, a sample is used for effect, and whether the sample has repair effect or not is evaluated by measuring the content of IL-1 beta and the like. In this embodiment, the repair treatment group: first add H 2 O 2 After 2h of stimulation, 100 μl of sample was added to each well for 24h of incubation;
the detection of the protective effect refers to that after a sample acts on cells, the cells are damaged again, the secretion conditions of IL-1 beta of the cells are observed, and whether the sample has the protective effect or not is judged. The protection processing group in this embodiment: adding sample, culturing for 24 hr, and adding H 2 O 2 Stimulation was performed for 2h.
The test indexes are as follows: IL-1 beta, IL-8 (test methods refer to kit instructions). The results are shown in FIGS. 1-8.
FIG. 1 is a schematic diagram showing the detection results of the anti-inflammatory effect of the protection treatment of the Phellinus linteus fermentate in example 2, the indexes: IL-1 beta. As can be seen from the graph, the IL-1β of the Model group (Model) is significantly higher than that of the blank Control group (Control), which indicates that the Model establishment is successful; 1% ZZ-TD was not significantly different from the model group, and 1% ZZ-TD-5372 was significantly lower than the model group after inoculation and fermentation, indicating that the fermentation broth after inoculation helped reduce IL-1β secretion, and had anti-inflammatory effects.
FIG. 2 is a schematic diagram showing the detection results of the anti-inflammatory effect of the protection treatment of the extract of the pearl potato in example 2, which is an index: IL-1 beta. From the graph, the IL-1 beta of the model group is extremely higher than that of the blank control group, which indicates that the model is successfully established, and the influence of the pearl potato extracts with different proportions on the IL-1 beta has larger variation. When the pearl proportion is 0, the effect of reducing IL-1 beta is not achieved, the effect of resisting inflammation is not achieved, the pearl potato extract added with 1% -5% of pearl powder has extremely remarkable effect of reducing IL-1 beta, the effect of resisting inflammation is achieved, and when the pearl adding proportion is increased to 10%, the effect on IL-1 beta is not remarkably different, and the effect of resisting inflammation is not achieved.
FIG. 3 is a schematic diagram showing the detection results of the anti-inflammatory effect of the protection treatment of the phellinus linteus pearl ferments with different concentrations in example 2, and the indexes are as follows: IL-1 beta. From the graph, the IL-1 beta of the model group is extremely higher than that of the blank control group, which means that the model is successfully established, 5372 is inoculated to a fermentation product in a potato culture medium without pearl addition, and the secretion of the IL-1 beta can be remarkably reduced, so that the anti-inflammatory effect is realized; when 1 to 5 percent of pearls are added according to the proportion, the anti-inflammatory effect is more remarkable; however, inoculation of 5372 when added to 10% pearls did not reduce IL-1 β.
FIG. 4 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the Phellinus linteus pearl fermentate in example 2, which is an index: IL-1 beta. From the figure, the IL-1 beta of the model group is extremely higher than that of the blank control group, which indicates that the model is successfully established, and the effect of potato extract with the same pearl addition proportion on the IL-1 beta is compared before and after fermentation, so that the fermentation product after 5372 inoculation has obvious effect of reducing the IL-1 beta, but has no effect of reducing the IL-1 beta before fermentation.
FIG. 5 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the extract of the potato in example 2, the indexes: IL-1 beta. From the graph, the IL-1 beta of the model group is extremely obviously higher than that of the blank control group, which indicates that the model is successfully established; the pearl potato extract with the pearl adding proportion of 5% has repairing effect, and the pearl potato extract with the pearl adding amount of 1% and the pearl adding amount of 10% has no effect of reducing IL-1 beta.
FIG. 6 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the phellinus linteus pearl ferments with different concentrations in example 2, and the indexes are as follows: IL-1 beta. From the graph, the IL-1 beta of the model group is extremely obviously higher than that of the blank control group, which indicates that the model is successfully established; the secretion of IL-1 beta can not be reduced in the potato culture medium without pearl addition, and the potato culture medium has no anti-inflammatory and repairing effects, no matter before fermentation or after fermentation; when 1 to 5 percent of pearls are added according to the proportion, the anti-inflammatory repairing effect is obvious; however, inoculation of 5372 fermentation products when added to 10% pearls did not reduce IL-1 beta.
FIG. 7 is a schematic diagram showing the detection results of the anti-inflammatory effect of the repair treatment of the extract of the potato in example 2, the index: IL-8. From the figure, the model group IL-8 is obviously higher than that of the blank control group, which indicates that the model establishment is successful; the pearl potato extract with the addition of 10% of pearls has the effect of reducing IL-8, namely has the repairing effect, but the potato extract with the addition of 1% and 5% of pearls has no anti-inflammatory effect.
FIG. 8 is a graph showing the detection results of the anti-inflammatory effect of the repair treatment of the phellinus linteus pearl ferments with different concentrations in example 2, and the indexes are as follows: IL-8. From the graph, the model group IL-8 is extremely higher than the blank control group, which indicates that the model establishment is successful; 5372 potato fermentation broth without pearl addition has the effect of reducing IL-8, which indicates that the potato fermentation broth has a repairing effect; the potato fermentation liquor with the pearl addition amount of 1-5% also has the effect of reducing IL-8, which indicates that the potato fermentation liquor has a repairing effect; when the addition amount was increased to 10%, there was no repair effect (no significant effect on IL-8).
2. Test of tyrosinase activity inhibition by Phellinus linteus fermentation product
1. Principle of experiment
The melanin is formed by gradually converting tyrosine in skin melanocyte tissue into eumelanin under the action of tyrosinase and other enzymes via dopa, dopaquinone, dopachrome, dihydroxyindole and other intermediates; furthermore, melanocyte tissue transfers melanin into keratin cells and falls off as the stratum corneum is periodically refreshed, and the difference in human skin color is mainly dependent on the content and distribution of eumelanin and pheomelanin, but when eumelanin grows excessively and is unevenly distributed, local skin hypermelanin or pigmentation is caused. There are three major enzymes involved in melanin synthesis: tyrosinase, dopachrome tautomerase and dihydroxyindole carboxylic acid oxidase. Tyrosinase is an oxidation-reduction enzyme, is a main speed-limiting enzyme for melanin synthesis, the activity of the tyrosinase is used for determining the quantity of melanin formation, and a plurality of whitening and freckle-removing products sold on the market at present are used for inhibiting the activity of tyrosinase to achieve the whitening effect, so that the inhibition effect on tyrosinase is a main index for evaluating whitening cosmetics. The L-tyrosinase and its substrate L-tyrosine can undergo catalytic reaction. When a reagent having an L-tyrosinase activity inhibiting effect is added to the experimental system, an inhibiting effect can be produced on the catalytic reaction, and the inhibition rate of the reagent on the L-tyrosinase activity can be evaluated by measuring absorbance at 475nm before and after the addition of the reagent.
2. Preparation of experiments
1) PBS phosphate buffer formulation (ph=6.8): 17.91g of disodium hydrogen phosphate dodecahydrate is taken and dissolved in distilled water to a constant volume of 500mL; dissolving 7.8g of sodium dihydrogen phosphate dihydrate in distilled water to a volume of 500mL; 92.6mL and 107.4mL of the two solutions were prepared as 200mL PBS phosphate buffer at pH 6.8.
2) 0.1mol/L hydrochloric acid solution: 0.862mL of HCl solution with 36-38% of HCl content is used, and distilled water is used for constant volume to 100mL.
3) 0.05% L-tyrosine solution: 0.05. 0.05g L-tyrosine was dissolved in 35mL of 0.1mol/L HCl solution, and 65mL of PBS phosphate buffer, pH6.8, was added thereto, with a total of 100mL.
4) Tyrosinase solution: the tyrosinase powder was diluted with PBS buffer to give a tyrosinase solution with an enzyme activity of 100U/mL.
3. Experimental procedure
1) After the C2 tube is uniformly mixed and shaken, the mixture is heated in a water bath for 10min in a water bath kettle at 37 ℃ and zeroed at the wavelength of 475 nm.
2) The C1 tube solution was prepared and shaken well, after 10min in a 37℃water bath, 1ml of tyrosinase was added, and the water bath was continued for 10min to determine the C1 absorbance.
3) The absorbance of T1 was measured by zeroing T2 in the same manner as in 1) and 2).
4) And calculating the activity inhibition ratio T (%) of the fermentation stock solution sample to tyrosinase.
TABLE 5 Experimental reagent proportioning table for tyrosinase inhibition rate
Numbering device | L-tyrosine | Sample of | PBS | Tyrosinase enzyme | Total volume of |
C1 | 2mL | —— | 4mL | 1mL | 7ml |
C2 | 2mL | —— | 5ml | —— | 7ml |
T1 | 2ml | 2mL | 2mL | 1mL | 7ml |
T2 | 2ml | 2ml | 3ml | —— | 7ml |
4. Data processing
The formula: t (%) = (C1-T1)/c1×100%. The results are shown in Table 6, and the data in the Table show that the fomes fomentarius and the pearl are fermented, and the fomes fomentarius and the pearl have synergistic effect and remarkably improve the whitening performance of the fermented product.
TABLE 6 tyrosinase inhibition assay results
Finally, it is further noted that in this disclosure relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.
While the disclosure has been disclosed by the foregoing description of specific embodiments thereof, it will be understood that various modifications, improvements, or equivalents may be devised by those skilled in the art that will fall within the spirit and scope of the appended claims. Such modifications, improvements, or equivalents are intended to be included within the scope of this disclosure.
Claims (9)
1. A method for preparing a fomes fomentarius ferment, comprising:
step one, carrying out activation, purification and expansion culture treatment on phellinus linteus to obtain seed liquid; the preservation number of the fomes fomentarius is CGMCC NO. 5.1494;
step two, inoculating the seed solution to a fermentation substrate for fermentation culture treatment to obtain fermentation liquor; the fermentation substrate is potato homogenate, and the potato homogenate also comprises 0.3 weight percent of monopotassium phosphate, 0.15 weight percent of magnesium sulfate, 0.05 weight percent of glucose and 0.2 weight percent of soybean peptide; the fermentation substrate is also added with pearl powder, and the content of the pearl powder is 1-10wt% of the potato homogenate; the temperature of the fermentation culture treatment is 25-35 ℃ and the time is 48-96 hours;
and thirdly, separating and sterilizing the fermentation liquor to obtain supernatant.
2. The method for producing a fermentation product of fomes fomentarius as claimed in claim 1, wherein the potato homogenate is prepared by homogenizing and dissolving fresh potato slices and water in a homogenizer, wherein the potato content is 1-5wt%.
3. The method of producing a fomes fomentarius ferment as claimed in claim 1, wherein the pearl powder is present in an amount of 1-5wt% of the potato homogenate.
4. A method for producing a fomes fomentarius fermentation product as claimed in any one of claims 1 to 3 wherein in step one, the hypha volume percentage of the seed liquid is 80% or more.
5. The method of producing a fermentation product of Phellinus linteus as set forth in claim 4, wherein in the second step, the volume ratio of the seed liquid to the fermentation substrate is 1% to 10%.
6. The method for producing a fomes fomentarius ferment according to claim 1, wherein in the second step, the fermentation culture treatment is carried out in a shaking table at a rotation speed of 150r/min to 200r/min.
7. The method for producing a fomes fomentarius ferment according to any one of claims 1 to 3, 5 to 6, wherein the separation treatment employs a centrifugation method.
8. The method for producing a fomes fomentarius fermentation product of claim 7, wherein the centrifugation speed is 4500r/min-5500r/min and the centrifugation time is 10min-20min.
9. A fomes fomentarius ferment, characterized in that it is produced according to the production process of any of claims 1-8; the fomes fomentarius fermentation product is fomes fomentarius fermentation liquid and dry powder thereof.
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