CN108066210B - A herba Hyperici Japonici leaf fermented product using domestic fungus, and its preparation method and application - Google Patents
A herba Hyperici Japonici leaf fermented product using domestic fungus, and its preparation method and application Download PDFInfo
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Abstract
The invention provides a herba rehmanniae praeparata leaf fermented product using edible fungi as well as a preparation method and application thereof, the herba rehmanniae praeparata leaf fermented product has good moisturizing effect, has no stimulation to skin, and can be used as a skin care product moisturizing agent; the technical scheme is as follows: fermenting herba Hyperici Japonici with edible fungus.
Description
Technical Field
The invention relates to a camellia sinensis fermentation product, in particular to a camellia sinensis fermentation product utilizing edible fungi, and also relates to a preparation method and application of the camellia sinensis fermentation product, belonging to the technical field of cosmetics.
Background
The fermentation refers to a process in which microorganisms decompose or synthesize organic substances using enzymes contained in the microorganisms, and representative fermentation microorganisms include Bacillus subtilis, Lactobacillus, Aspergillus, yeast, mold, Acetobacter, Agaricus campestris mycelium, and the like. Recently, in the field of cosmetics, there has been an increasing interest in the production of fermented foods and cosmetics from natural materials, and studies on whitening, improvement of skin wrinkles, oxidation resistance, moisture retention, elasticity enhancement, and the like have been widely conducted.
The fermented product or its extract is prepared by fermenting and ripening to convert sugar in natural materials into alcohol, and can improve extraction efficiency of effective components such as flavonoid, vitamin C, vitamin P, amino acids, and catechin. In addition, sometimes natural materials having irritation or toxicity are stabilized or reduced in toxicity or converted into stable derivatives by fermentation extraction, and the cosmetic efficacy can be greatly improved. In addition, in the process of converting into easily absorbable low molecular substances through fermentation processes of lactic acid bacteria, yeast and the like, and converting from unstable or inert forms into active forms, the effects of natural materials can be greatly improved, and the effects of moisturizing, elasticity enhancing, whitening and skin wrinkle improving can be enhanced.
The edible fungi contain abundant protein and amino acid, and the content of the edible fungi is several times to dozens of times of that of common vegetables and fruits. The edible fungus contains 18 kinds of amino acids for constituting protein and 8 kinds of trace elements essential for human body. The cereal food contains less lysine and the edible fungi are rich. The edible fungi has low fat content, which is 0.2-3.6 wt% of dry product, and 74-83% of unsaturated fatty acid beneficial to human health. The edible fungi also contain vitamins, and the vitamins are rich in VB1 and V12 which are higher than meat; the original content of the mushroom Vd is up to 128 international units, which is 8 times of that of the laver. The edible fungi are also rich in various mineral elements such as phosphorus, potassium, sodium, calcium, iron, zinc, magnesium, manganese and the like and other trace elements.
The edible fungi contain bioactive substances such as high molecular polysaccharide, beta-glucose and RNA complex, natural organic germanium, nucleic acid degradation products, cAMP and triterpenoids, and the like, and have important utilization value for maintaining human health. The edible fungus has the medicinal and health-care values of resisting cancer, stimulating the formation of antibodies, improving and adjusting the internal defense capacity of an organism, reducing the incidence rate of certain substances inducing tumors and having synergistic effect on various chemotherapeutic drugs. In addition, the chestnut mushroom is rich in organic selenium, can be used as a selenium supplement food, and can almost prevent all canceration if eaten for a long time. ② antibacterial and antiviral effects. ③ lowering blood pressure, reducing blood fat, resisting thrombus, resisting arrhythmia, strengthening heart, etc. Fourthly, the stomach-invigorating and digestion-aiding functions. Fifthly, relieving cough and asthma and eliminating phlegm. Sixthly, cholagogue, liver protection and detoxification. And lowering blood sugar. The prescription is also provided with a preparation method. Ninthly, immunoregulation. Health food, health drink, wine and medicine produced and processed by taking edible fungi as raw materials are used in medical clinic and put into the health product market in large quantity.
The herba rehmanniae is also named as white snow tea and Taibai tea, is shaped like white chrysanthemum petals, is white like snow, so the Lijiang snow tea is named as Lijiang Yulong snow tea, and grows in a snow region alpine moss plant zone with the altitude of more than 4000 meters in Lijiang Yulong snow mountain. The chemical components of the snow tea have very good health care effect. The chemical components of the material comprise 3.5-7.0% of inorganic matters and 93-96.5% of organic matters. About 27 inorganic mineral elements in tea include phosphorus, potassium, sulfur, magnesium, manganese, fluorine, aluminum, calcium, sodium, iron, copper, zinc, selenium, etc. The organic compounds in folium Camelliae sinensis mainly include protein, lipid, carbohydrate, amino acids, alkaloids, tea polyphenols, organic acids, pigment, aroma components, vitamins, saponin, sterol, etc. The herba Hyperici Japonici has effects of reducing blood lipid and blood pressure, reducing weight, refreshing brain, moistening lung, clearing summerheat, promoting fluid production, relieving cough, removing liver fire, improving eyesight, removing toxin, suppressing hyperactive liver, lowering fire, nourishing yin, moistening lung, and can be used for treating pneumonia cough, hypertension, neurasthenia, pharyngolaryngitis, mania, dysphoria, etc. However, the research on the application of the Japanese ardisia herb to cosmetics is relatively small.
Disclosure of Invention
Therefore, the invention provides the camellia sinensis fermented product fermented by the edible fungi, which has a good moisturizing effect, has no stimulation to skin, can be used as a skin care product moisturizing agent, and can effectively improve the problems of dry, tight and fine lines of skin.
In order to achieve the above object, a first technical solution provided by the present invention is as follows:
a herba Thymi Japonici fermented product is prepared by fermenting herba Thymi Japonici with edible fungus.
Further, the above-mentioned fermented product of herba schizonepetae tea using edible fungi comprises Sparassis crispa, Grifola frondosa and Tricholoma matsutake.
Furthermore, the camellia sinensis fermentation product utilizing the edible fungi also comprises the truffle.
Furthermore, the camellia sinensis leaf fermented product utilizing the edible fungi also comprises the tuber summer truffle.
The second technical scheme provided by the invention is as follows:
further, the preparation method of the camellia sinensis fermentation product by using the edible fungi sequentially comprises the following steps of:
(1) preparation of seed liquid
Respectively shearing each edible fungus fruiting body under aseptic condition to obtain tissue segments, respectively inoculating to slant culture medium, and standing at 25-28 deg.C for 8-10 days; selecting 10-20 pieces of agar with hyphae from a slant culture medium under the aseptic condition, inoculating the agar into the aseptic slant culture medium again, performing dark culture at the temperature of 25-28 ℃ for 12-15 days, then performing illumination for 3-4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the mother strain into a seed culture medium, and performing culture for 5-7 days under the conditions that the temperature is 26-28 ℃ and the rotating speed of a shaking table is 150 plus materials at 200r/min to respectively obtain seed solutions of the edible fungi;
(2) ultrafine pulverizing herba Hyperici Japonici to 80-100 μm, adding 2-4 times of water, soaking at 30-40 deg.C for 1-2 days, and centrifuging to obtain herba Hyperici Japonici soaking extract;
(3) mixing the edible fungus seed liquids prepared in the step (1) according to an equal volume ratio in advance, inoculating 5-10% of inoculum size into a 5L fermentation tank of a high-density fermentation culture medium, adding the herba Hyperici Japonici soaking extract in the step (2) to start fermentation culture, wherein the mass ratio of the herba Hyperici Japonici soaking extract to the edible fungus seed liquids is 3-6: 1, the temperature is 28-30 ℃, the rotating speed is 100-250r/min, and the ventilation volume is 0.5-1.5dm3Fermenting and culturing for 5-10 days at min, performing submerged fermentation to obtain mixture of edible fungus fermented tea, centrifuging, collecting supernatant, and filtering.
(4) And (4) concentrating the filtrate obtained by filtering in the step (3) under reduced pressure at the temperature of below 60 ℃ until the water content is lower than 8%, and then freeze-drying to obtain powder, namely the camellia sinensis fermented product.
Further, in the above preparation method of a camellia sinensis leaf fermented product using edible fungi, the slant culture medium in the step (1) is a PDA culture medium or a PDPA comprehensive culture medium.
Further, in the above method for preparing a fermented product of camellia sinensis leaves by using edible fungi, the seed culture medium in the step (1) is: pH6.0-6.5, calculated by 100% of total mass fraction, composed of glucose 1.5-2.5%, peptone 0.1-0.2%, beef extract 0.1-0.2%, KH2PO40.1-0.15％、MgSO40.1-0.15%, vitamin B10.01-0.02%, and water in balance.
Further, in the above preparation method of a fermented product of herba Hyperici Japonici using edible fungi, the pH of the fermentation medium in step (3) is 5.5-6.0, and the fermentation medium comprises, based on 100% of the total mass fraction, glucose-3%, peptone 0.4-0.6%, yeast extract 0.2-0.4%, and KH2PO40.15-0.3％、MgSO40.1-0.15%, vitamin B10.01-0.02%, and water in balance.
The last technical scheme of the invention is to provide the application of the camellia sinensis fermented product of the edible fungi as the moisturizing agent of the skin care product, wherein the moisturizing composition accounts for 0.1-20% of the skin care product by mass.
Compared with the prior art, the technical scheme provided by the invention has the following technical advantages:
1. the camellia sinensis leaf fermentation product is obtained by fermenting mycelium of edible fungi, organic solvents are removed in the production process, negative effects of the organic solvents on human bodies are avoided, and components sensitive to heat are reserved by adopting low-temperature fermentation; the high molecular compounds of the tea leaves are converted into low molecular compounds under the action of enzymes generated in the fermentation process, the obtained nutrient components are easy to permeate the skin, and the active ingredients of the edible fungi are combined, so that substances beneficial to the skin are added, and the tea leaves have good moisturizing and hydrating effects. The fermentation product can be applied to skin care cosmetics such as eye cream, face cream, facial mask, emulsion, gel, essence and the like, can bring good moisturizing effect to the products, and effectively improve the problems of dry, tight and fine lines of skin.
2. The technical scheme that this application provided utilizes multiple domestic fungus mixed fermentation ground tealeaves, and multiple domestic fungus can produce the enzyme system of different performance, and several kinds of domestic fungus are mutually favorable to be lived altogether, makes the enzyme system proportion coordinate, makes the zymolyte decompose more thoroughly, produces multiple small molecule active product, and these active product synergy reach better moisturizing effect.
Detailed Description
The invention will now be further described with reference to the following examples, which are not to be construed as limiting the invention in any way, and any limited number of modifications which can be made within the scope of the claims of the invention are still within the scope of the claims of the invention.
Example 1
The invention provides a fermented product of herba abri Japonici by edible fungi, which is obtained by fermenting herba abri Japonici with Sparassis crispa, Grifola frondosa and Tricholoma matsutake.
The above herba Hyperici Japonici fermented product is prepared by the following steps:
(1) preparing edible fungus seed liquid;
shearing Sparassis crispa fruiting body under aseptic condition to obtain tissue fragments, inoculating to PDA culture medium, and standing at 28 deg.C for 8 days; selecting 20 agar blocks with hyphae from a PDA culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 25 ℃ for 15 days, then performing illumination for 3 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 150r/min to obtain the sparassis crispa seed solution;
shearing the sporophore of Grifola frondosa under aseptic condition to obtain tissue fragment, inoculating to PDA culture medium, and standing at 28 deg.C for 8 days; selecting 20 agar blocks with hyphae from a PDA culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 25 ℃ for 15 days, then performing illumination for 3 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 150r/min to obtain the grifola frondosa seed solution;
shearing the Tricholoma matsutake fruiting body under aseptic condition to obtain tissue segment, inoculating to PDA culture medium, and standing at 28 deg.C for 8 days; selecting 20 agar blocks with hyphae from a PDA culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 25 ℃ for 15 days, then performing illumination for 3 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 150r/min to obtain the songi mushroom seed solution;
the seed culture medium in the step (1) is as follows: pH6.0, calculated by the total mass fraction of 100%, composed of 2.5% of glucose, 0.1% of peptone, 0.2% of beef extract and KH2PO40.1％、MgSO40.15 percent, 10.01 percent of vitamin B and the balance of water.
(2) Ultrafine pulverizing herba Hyperici Japonici to 100 μm, adding 2 times of water, soaking at 40 deg.C for 1 day, and centrifuging to obtain herba Hyperici Japonici soaking extract;
(3) various kinds prepared in the step (1)The volume ratio of the seed liquid to the mother liquid is 1: 1: 1, mixing, inoculating 10% of inoculum size into a 5L fermentation tank of a high-density fermentation medium, adding the herba Hyperici Japonici soaking extract obtained in the step (2), and starting fermentation culture, wherein the mass ratio of the herba Hyperici Japonici soaking extract to the edible fungus seed liquid is 3: 1, at a temperature of 30 ℃, a rotation speed of 100r/min and an air flow of 1.5dm3Fermenting and culturing for 5 days at min, performing submerged fermentation to obtain mixture of edible fungus fermented tea, centrifuging, collecting supernatant, and filtering.
The pH value of the fermentation medium in the step (3) is 5.5, and the fermentation medium is composed of 3% of glucose, 0.4% of peptone, 0.4% of yeast extract and KH according to 100% of total mass fraction2PO40.15％、MgSO40.15 percent, 10.01 percent of vitamin B and the balance of water.
(4) And (4) carrying out reduced pressure concentration on the filtrate obtained by filtering in the step (3) at the temperature of 60 ℃ until the water content is 8%, and then carrying out freeze drying to obtain powder, namely the camellia sinensis fermented product.
Example 2
The invention provides a fermented product of herba pachyrhizus using edible fungi, which is obtained by fermenting herba pachyrhizus using summer truffle, sparassis crispa, grifola frondosa and tricholoma matsutake.
The above herba Hyperici Japonici fermented product is prepared by the following steps:
(1) preparing edible fungus seed liquid
Shearing Sparassis crispa fruiting body under aseptic condition to obtain tissue fragment, inoculating to PDPA comprehensive culture medium, and standing at 25 deg.C for 10 days; selecting 10 pieces of agar with hyphae from a PDPA comprehensive culture medium under aseptic conditions, inoculating the agar into the aseptic PDPA comprehensive culture medium again, performing dark culture at 28 ℃ for 12 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the strain into a seed culture medium, and performing culture for 5 days at the temperature of 26 ℃ and the rotating speed of a shaking table of 200r/min to obtain a sparassis crispa seed solution;
shearing the sporophore of Grifola frondosa under aseptic condition to obtain tissue fragment, inoculating to PDPA comprehensive culture medium, and standing at 25 deg.C for 10 days; selecting 10 pieces of agar with hyphae from a PDPA comprehensive culture medium under aseptic conditions, inoculating into the aseptic PDPA comprehensive culture medium again, performing dark culture at 28 ℃ for 12 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 5 days at the temperature of 26 ℃ and the rotating speed of a shaking table of 200r/min to obtain a grifola frondosa seed solution;
shearing the sporophore of Tricholoma matsutake under aseptic condition to obtain tissue fragment, inoculating to PDPA comprehensive culture medium, and standing at 25 deg.C for 10 days; selecting 10 pieces of agar with hyphae from a PDPA comprehensive culture medium under aseptic conditions, inoculating the agar into the aseptic PDPA comprehensive culture medium again, performing dark culture at 28 ℃ for 12 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the strain into a seed culture medium, and performing culture for 5 days at the temperature of 26 ℃ and the rotating speed of a shaking table of 200r/min to obtain a songi mushroom seed solution;
shearing the fruit body of the summer truffle under the aseptic condition to obtain tissue segments, inoculating the tissue segments to a PDPA comprehensive culture medium, and standing and culturing for 10 days at the temperature of 25 ℃; selecting 10 pieces of agar with hyphae from a PDPA comprehensive culture medium under aseptic conditions, inoculating the agar into the aseptic PDPA comprehensive culture medium again, culturing in dark at 28 ℃ for 12 days, then illuminating for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the strain into a seed culture medium, and culturing for 5 days at 26 ℃ and the rotating speed of a shaking table of 200r/min to obtain a tuber muskmelon seed solution;
the seed culture medium in the step (1) is as follows: pH6.0, calculated by the total mass fraction of 100%, composed of 1.5% of glucose, 0.2% of peptone, 0.1% of beef extract and KH2PO40.15％、MgSO40.1 percent, 10.02 percent of vitamin B and the balance of water.
(2) Ultrafine pulverizing herba Hyperici Japonici to 80 μm, adding 4 times of water, soaking at 30 deg.C for 2 days, and centrifuging to obtain herba Hyperici Japonici soaking extract;
(3) pre-mixing each seed solution prepared in the step (1) according to a volume ratio of 1: 1: 1: 1, mixing, inoculating 5% of inoculum size into 5L fermentation tank of high density fermentation medium, adding herba Hyperici Japonici soaking extractive solution of step (2) to start fermentation culture,the mass ratio of the ground tea leaf soaking extract to the edible fungus seed liquid is 6: 1, at a temperature of 28 ℃, a rotation speed of 250r/min and an air flow of 0.5dm3Fermenting and culturing for 10 days at min, performing submerged fermentation to obtain mixture of edible fungus fermented tea, centrifuging, collecting supernatant, and filtering.
The pH value of the fermentation medium in the step (3) is 5.5-6.0, and the fermentation medium is composed of 2% of glucose, 0.6% of peptone, 0.2% of yeast extract and KH (KH) according to 100% of the total mass fraction2PO40.3％、MgSO40.1 percent, 10.02 percent of vitamin B and the balance of water.
(4) And (4) carrying out reduced pressure concentration on the filtrate obtained by filtering in the step (3) at 50 ℃ until the water content is 5%, and then carrying out freeze drying to obtain powder, namely the camellia sinensis fermented product.
Example 3
The invention provides a fermented product of herba pachyrhizus using edible fungi, which is obtained by fermenting herba pachyrhizus using white fungus, Sparassis crispa, Grifola frondosa and Tricholoma matsutake.
The above herba Hyperici Japonici fermented product is prepared by the following steps:
(1) preparing edible fungus seed liquid
Shearing Sparassis crispa fruiting body under aseptic condition to obtain tissue fragment, inoculating to PDPA comprehensive culture medium, and standing at 26 deg.C for 9 days; selecting 15 agar blocks with hyphae from a PDPA comprehensive culture medium under a sterile condition, inoculating the agar blocks into a sterile PDA culture medium or a PDPA comprehensive culture medium again, performing dark culture at 26 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the strain into a seed culture medium, and performing culture for 6 days under the conditions that the temperature is 27 ℃ and the rotating speed of a shaking table is 180r/min to obtain the Sparassis crispa seed solution;
shearing tissue segments of grifola frondosa sporocarp under an aseptic condition, inoculating the tissue segments to a PDPA comprehensive culture medium, carrying out static culture at the temperature of 26 ℃ for 9 days, selecting 15 agar blocks with hyphae from the PDPA comprehensive culture medium under the aseptic condition, inoculating the agar blocks into a sterile PDA culture medium or PDPA comprehensive culture medium again, carrying out dark culture at the temperature of 26 ℃ for 14 days, illuminating for 4 days, selecting a strain which is fast in color change and regular in hyphae growth as a mother strain, then inoculating the mother strain into a seed culture medium, and culturing for 6 days at the temperature of 27 ℃ and the rotating speed of a shaking table of 180r/min to obtain an edible fungus seed solution;
shearing the sporophore of Tricholoma matsutake under aseptic condition to obtain tissue fragment, inoculating to PDPA comprehensive culture medium, and standing at 26 deg.C for 9 days; selecting 15 agar blocks with hyphae from a PDPA comprehensive culture medium under a sterile condition, inoculating the agar blocks into a sterile PDA culture medium or a PDPA comprehensive culture medium again, performing dark culture at 26 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the strain into a seed culture medium, and performing culture for 6 days under the conditions that the temperature is 27 ℃ and the rotating speed of a shaking table is 180r/min to obtain the Tricholoma matsutake seed solution;
shearing the fruiting body of the truffle under aseptic condition to obtain tissue segments, inoculating to PDPA comprehensive culture medium, and standing and culturing at 26 deg.C for 9 days; selecting 15 agar blocks with hyphae from a PDPA comprehensive culture medium under the aseptic condition, inoculating the agar blocks into a sterile PDA culture medium or a PDPA comprehensive culture medium again, performing dark culture at 26 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the strain into a seed culture medium, and performing culture for 6 days under the conditions that the temperature is 27 ℃ and the rotating speed of a shaking table is 180r/min to obtain the chrysosporium leucatum seed solution;
the seed culture medium in the step (1) is as follows: pH6.3, calculated by the total mass fraction of 100%, composed of 2% of glucose, 0.15% of peptone, 0.15% of beef extract and KH2PO40.12％、MgSO40.12 percent of vitamin B, 10.015 percent of vitamin B and the balance of water.
(2) Ultrafine pulverizing herba Hyperici Japonici to 90 μm, adding 3 times of water, soaking at 35 deg.C for 1.5 days, and centrifuging to obtain herba Hyperici Japonici soaking extract;
(3) pre-mixing each seed solution prepared in the step (1) according to a volume ratio of 1: 1: 1: 1, mixing, inoculating 7% of inoculum size into a 5L fermentation tank of a high-density fermentation medium, adding the herba Hyperici Japonici soaking extract obtained in the step (2), and starting fermentation culture, wherein the mass ratio of the herba Hyperici Japonici soaking extract to the edible fungus seed liquid is 5: 1At a temperature of 28 deg.C, a rotation speed of 200r/min and a ventilation amount of 1.0dm3Fermenting and culturing for 8 days at min, performing submerged fermentation to obtain mixture of edible fungus fermented tea, centrifuging, collecting supernatant, and filtering.
The pH value of the fermentation medium in the step (3) is 5.8, and the fermentation medium comprises 2.5 percent of glucose, 0.5 percent of peptone, 0.25 percent of yeast extract and KH according to 100 percent of total mass fraction2PO40.2％、MgSO40.12 percent of vitamin B, 10.015 percent of vitamin B and the balance of water.
(4) And (4) concentrating the filtrate obtained by filtering in the step (3) under reduced pressure at 55 ℃ until the water content is 6%, and then freeze-drying to obtain powder, namely the camellia sinensis fermented product.
Example 4
The invention provides a camellia sinensis leaf fermented product using edible fungi, which is obtained by fermenting camellia sinensis leaves by using fungus tuber, white truffle, sparassis crispa, grifola frondosa and tricholoma matsutake.
The above herba Hyperici Japonici fermented product is prepared by the following steps:
(1) preparing edible fungus seed liquid
Shearing Sparassis crispa fruiting body under aseptic condition to obtain tissue fragments, inoculating to PDA culture medium, and standing at 27 deg.C for 9 days; selecting 18 agar blocks with hyphae from a PDA culture medium or a PDPA comprehensive culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 27 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 170r/min to obtain the edible fungus seed liquid;
shearing the fruiting body of the truffle under aseptic condition to obtain tissue segments, inoculating to PDA culture medium, and standing at 27 deg.C for 9 days; selecting 18 agar blocks with hyphae from a PDA culture medium or a PDPA comprehensive culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 27 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 170r/min to obtain the white truffle seed solution;
shearing the sporophore of Grifola frondosa under aseptic condition to obtain tissue fragment, inoculating to PDA culture medium, and standing at 27 deg.C for 9 days; selecting 18 agar blocks with hyphae from a PDA culture medium or a PDPA comprehensive culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 27 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 170r/min to obtain the grifola frondosa seed solution;
shearing the sporophore of Tricholoma matsutake under aseptic condition to obtain tissue fragment, inoculating to PDA culture medium, and standing at 27 deg.C for 9 days; selecting 18 agar blocks with hyphae from a PDA culture medium or a PDPA comprehensive culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 27 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 170r/min to obtain a songi mushroom seed solution;
shearing the fruiting body of the summer truffle under aseptic condition to obtain tissue segments, inoculating the tissue segments to a PDA culture medium, and standing and culturing for 9 days at the temperature of 27 ℃; selecting 18 agar blocks with hyphae from a PDA culture medium or a PDPA comprehensive culture medium under the aseptic condition, inoculating into the aseptic PDA culture medium again, performing dark culture at 27 ℃ for 14 days, then performing illumination for 4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating into a seed culture medium, and performing culture for 7 days at the temperature of 28 ℃ and the rotating speed of a shaking table of 170r/min to obtain a summer truffle seed solution;
the seed culture medium in the step (1) is as follows: pH6.2, calculated by the total mass fraction of 100%, composed of 2% of glucose, 0.16% of peptone, 0.16% of beef extract and KH2PO40.13％、MgSO40.13 percent of vitamin B10.016 percent and the balance of water.
(2) Ultrafine pulverizing herba Hyperici Japonici to 90 μm, adding 3 times of water, soaking at 36 deg.C for 1.5 days, and centrifuging to obtain herba Hyperici Japonici soaking extract;
(3) pre-mixing each seed solution prepared in the step (1) according to a volume ratio of 1: 1: 1: 1: 1, mixing, inoculating 7% of inoculum size into a 5L fermentation tank of a high-density fermentation medium, adding the herba Hyperici Japonici soaking extract obtained in the step (2), and starting fermentation culture, wherein the mass ratio of the herba Hyperici Japonici soaking extract to the edible fungus seed liquid is 4: 1, at a temperature of 30 ℃, a rotation speed of 150r/min and an air flow of 0.8dm3Fermenting and culturing for 7 days at min, performing submerged fermentation to obtain mixture of edible fungus fermented tea, centrifuging, collecting supernatant, and filtering.
The pH value of the fermentation medium in the step (3) is 5.9, and the fermentation medium comprises 2.4 percent of glucose, 0.6 percent of peptone, 0.3 percent of yeast extract and KH according to 100 percent of total mass fraction2PO40.18％、MgSO40.13 percent of vitamin B10.016 percent and the balance of water.
(4) And (4) carrying out reduced pressure concentration on the filtrate obtained by filtering in the step (3) at the temperature of 60 ℃ until the water content is 8%, and then carrying out freeze drying to obtain powder, namely the camellia sinensis fermented product.
The ground tea fermentation product provided by the invention is prepared by mixing various edible fungi including Sparassis crispa, Grifola frondosa, Tricholoma matsutake and summer truffle to ferment ground tea, wherein the various edible fungi can generate enzyme systems with different performances, and the edible fungi can mutually benefit and coexist, so that the enzyme systems are coordinated in proportion, fermentation substrates are decomposed more thoroughly, various small molecular active products are generated, and the active products are synergized to achieve better moisturizing and water replenishing effects.
The camellia sinensis fermented product fermented by the edible fungi can be used as a skin care product additive and added into skin care products in various dosage forms such as eye cream, face cream, facial mask, emulsion, gel, essence and the like, so that the product has better effects of moisturizing and supplementing water, and the problems of dry skin, tightness and fine lines of the skin are effectively solved; the composition in the skin care product is 0.1-20% by mass.
Comparative example 1 preparation of common Dicha leaf extract
The tea extract is prepared by adopting a conventional hot reflux extraction process, and the method comprises the following steps: ultrafine pulverizing herba Hyperici Japonici to 90 μm, adding 10 times of water, heating at 70 deg.C for 5 hr for hot reflux extraction, separating supernatant from precipitate, repeatedly extracting for 3 times, filtering extract, concentrating under reduced pressure at 55 deg.C, and freeze drying to obtain powder.
Comparative example 2 preparation of fermented Dicha leaf Using Sparassis crispa
The preparation method and fermentation culture method of Sparassis crispa seed solution are the same as in example 4.
Comparative example 3 preparation of fermented product of Dicha leaf from Grifola frondosa
The preparation method and fermentation culture method of Grifola frondosa seed liquid are the same as those of example 4.
Comparative example 4 preparation of fermented Dicha leaf Using Tricholoma matsutake
The preparation method and fermentation culture method of Tricholoma matsutake seed liquid are the same as those of example 4.
Comparative example 5 preparation of fermented product of Dicha leaves Using Armillaria Albizziae
The preparation method and fermentation culture method of the truffle seed liquid are the same as those of example 4.
Comparative example 6 preparation of fermented product of Dicha leaf Using Tuber Prunellae
The preparation method and fermentation culture method of the truffle seed liquid are the same as those of example 4.
Experimental example 1 moisture retention Performance test
1. The test population: 60 subjects with dry skin, age 20-60 years, were randomized sex and divided into 12 groups of 5 individuals each.
2. Preparation of test samples: the fermented products of camellia sinensis leaves prepared in examples 1 to 4, the extract of camellia sinensis leaves prepared in comparative example 1, the fermented products of camellia sinensis leaves prepared in comparative examples 2 to 6, and sodium hyaluronate were prepared into a 1% aqueous solution with distilled water.
2. The test method comprises the following steps:
1) the technician uses the skin moisture content test Corneometer CM825 to measure the test site of the subject, and the average is taken and recorded as a blank value.
2) Subjects were measured 5 times by a technician using a Corneometer CM825 for skin moisture content testing after 0.5h, 1h, 2h, 4h, 6h of continuous use of the test samples, averaged, and the values recorded, the results are shown in table 1;
TABLE 1 moisture content test results
| Test sample | 0h | 0.5h | 1h | 2h | 4h | 6h |
| Example 1 fermented product of Dicha | 35.22 | 64.38 | 56.04 | 51.79 | 49.71 | 49.31 |
| Example 2 tea fermentation | 36.81 | 69.53 | 60.98 | 56.68 | 54.85 | 54.32 |
| Example 3 tea fermentation | 35.65 | 67.88 | 59.21 | 55.67 | 53.45 | 52.71 |
| Example 4 tea fermentation | 34.15 | 68.74 | 61.48 | 56.35 | 54.38 | 53.86 |
| Comparative example 1 extract of Dicha leaves | 34.43 | 51.23 | 44.52 | 41.01 | 39.60 | 38.92 |
| Comparative example 2 fermented product of tea leaves | 36.82 | 61.68 | 52.79 | 49.92 | 47.08 | 46.10 |
| Comparative example 3 fermented product of tea leaves | 37.90 | 61.45 | 53.54 | 50.87 | 48.03 | 47.22 |
| Comparative example 4 fermented product of tea leaves | 37.08 | 61.63 | 50.98 | 48.53 | 46.75 | 45.76 |
| Comparative example 5 fermented product of tea leaves | 36.72 | 61.21 | 50.68 | 48.19 | 46.67 | 45.82 |
| Comparative example 6 fermented product of tea leaves | 37.90 | 62.39 | 52.72 | 48.97 | 47.43 | 46.42 |
| Hyaluronic acid sodium salt | 37.08 | 58.43 | 50.38 | 47.23 | 45.85 | 44.78 |
| Distilled water | 36.72 | 37.11 | 36.68 | 36.59 | 36.67 | 36.42 |
The applicant calculated the skin hydration rate after using the test sample according to the formula { (test value-blank value)/blank value } × 100%, and the results are shown in table 2.
Table 2 hydration test results
| Test sample | 0h | 0.5h | 1h | 2h | 4h | 6h |
| Example 1 fermented product of Dicha | 0 | 82.79 | 59.11 | 47.05 | 41.14 | 40.01 |
| Example 2 tea fermentation | 0 | 88.89 | 65.66 | 53.98 | 49.01 | 47.57 |
| Example 3 tea fermentation | 0 | 90.41 | 66.09 | 56.16 | 49.93 | 47.85 |
| Example 4 tea fermentation | 0 | 101.29 | 80.03 | 65.01 | 59.24 | 57.72 |
| Comparative example 1 extract of Dicha leaves | 0 | 48.79 | 29.31 | 19.11 | 15.02 | 13.04 |
| Comparative example 2 fermented product of tea leaves | 0 | 67.52 | 43.37 | 35.58 | 27.87 | 25.20 |
| Comparative example 3 fermented product of tea leaves | 0 | 62.14 | 41.27 | 34.22 | 26.73 | 24.59 |
| Comparative example 4 fermented product of tea leaves | 0 | 66.21 | 37.49 | 30.88 | 26.08 | 23.41 |
| Comparative example 5 fermented product of tea leaves | 0 | 66.69 | 38.02 | 31.24 | 27.10 | 24.78 |
| Comparative example 6 fermented product of tea leaves | 0 | 64.62 | 39.10 | 29.21 | 25.15 | 22.48 |
| Hyaluronic acid sodium salt | 0 | 57.58 | 35.87 | 27.37 | 23.65 | 20.77 |
| Distilled water | 0 | 1.06 | -0.11 | -0.35 | -0.14 | -0.82 |
The above results show that the fermented products of the camellia sinensis leaves of examples 1 to 4 of the present invention all have good moisturizing properties. After the fermented herba schizonepetae tea is used for 6 hours, the moisture content value of the skin is more than 49, the skin hydration rate is more than 40%, and the effect of keeping moisture of the fermented herba schizonepetae tea prepared by the invention is proved to be lasting.
In particular, the water content of the fermented product of camellia sinensis leaves prepared in example 4 was significantly higher than that of other examples 1 to 3, comparative examples 1 to 6, and sodium hyaluronate and distilled water. Therefore, the best moisturizing performance is achieved by adopting the camellia sinensis leaf fermented product obtained by mixing and fermenting the sparassis crispa, the grifola frondosa, the tricholoma matsutake, the white truffle and the summer truffle.
Experimental example 2 Observation on clinical trial
1) Testing the population: subjects aged 20-50 years, dry, tight and striae were divided into 10 groups of 200 persons. 20 people per group.
Experimental samples: the fermented herba desmodii dibotrys provided in examples 1-4 of the application, the herba desmodii dibotrys extract prepared in comparative example 1 and the fermented herba desmodii dibotrys prepared in comparative examples 2-6 are added into an essence base, and the addition amount of the fermented herba desmodii dibotrys is 10% of the total mass of the skin care product. The moisturizing essence liquid is named as the moisturizing essence liquid in the examples 1 to 4 and the moisturizing essence liquid in the comparative examples 1 to 6 in sequence.
3) The using method comprises the following steps: the test specimens were rubbed on the face after cleaning the face every morning and evening for 2 months.
4) As a result: more than 85% of users showed that the skin was dry, tight and fine lines were significantly improved after using the moisturizing essence of examples 1-4 for 2 months, wherein more than 95% of users showed that the skin became moist, smooth and fine after using the moisturizing essence of example 4 for 2 months; while the moisturizing essence of comparative examples 1 to 6 was inferior to that of inventive examples 1 to 4, only 40% of users showed improvement of dry skin after 2 months using the moisturizing essence of example 1.
5) After the test sample prepared by the invention is insisted on the test sample for 3 months, the test sample does not have allergic symptoms such as face pain and itch, red swelling, edema, inflammation and the like.
In conclusion, the camellia sinensis leaf fermented product utilizing the edible fungi has a good moisturizing effect, can effectively solve the problems of dry skin, tightness and fine lines, and cannot cause skin allergy.
Claims (7)
1. A herba Blumeae Laciniatae fermented product using edible fungi is prepared by fermenting herba Blumeae Laciniatae, Maitake Mushroom, Tricholoma matsutake, herba Agastaches, and summer truffle to obtain herba Blumeae Laciniatae fermented product as skin care product humectant;
the camellia sinensis leaf fermented product utilizing the edible fungi is prepared by the following method in sequence:
(1) preparation of seed liquid
Respectively shearing each edible fungus fruiting body under aseptic condition to obtain tissue segments, respectively inoculating to slant culture medium, and standing at 25-28 deg.C for 8-10 days; selecting 10-20 pieces of agar with hyphae from a slant culture medium under the aseptic condition, inoculating the agar into the aseptic slant culture medium again, performing dark culture at the temperature of 25-28 ℃ for 12-15 days, then performing illumination for 3-4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the mother strain into a seed culture medium, and performing culture for 5-7 days under the conditions that the temperature is 26-28 ℃ and the rotating speed of a shaking table is 150 plus materials at 200r/min to respectively obtain seed solutions of the edible fungi;
(2) ultrafine pulverizing herba Hyperici Japonici to 80-100 μm, adding 2-4 times of water, soaking at 30-40 deg.C for 1-2 days, and centrifuging to obtain herba Hyperici Japonici soaking extract;
(3) mixing the edible fungus seed liquids prepared in the step (1) according to an equal volume ratio in advance, inoculating 5-10% of inoculum size into a 5L fermentation tank of a high-density fermentation culture medium, adding the herba Hyperici Japonici soaking extract in the step (2) to start fermentation culture, wherein the mass ratio of the herba Hyperici Japonici soaking extract to the edible fungus seed liquids is 3-6: 1, the temperature is 28-30 ℃, the rotating speed is 100-250r/min, and the ventilation volume is 0.5-1.5dm3Fermenting and culturing for 5-10 days at min to obtain mixture of edible fungus fermented tea, centrifuging, and filtering supernatant;
(4) and (4) concentrating the filtrate obtained by filtering in the step (3) under reduced pressure at the temperature of below 60 ℃ until the water content is lower than 8%, and then freeze-drying to obtain powder, namely the camellia sinensis fermented product.
2. The method for preparing a fermented product of herba Hyperici Japonici using edible fungi of claim 1, which comprises the following steps in order:
(1) preparation of seed liquid
Respectively shearing each edible fungus fruiting body under aseptic condition to obtain tissue segments, respectively inoculating to slant culture medium, and standing at 25-28 deg.C for 8-10 days; selecting 10-20 pieces of agar with hyphae from a slant culture medium under the aseptic condition, inoculating the agar into the aseptic slant culture medium again, performing dark culture at the temperature of 25-28 ℃ for 12-15 days, then performing illumination for 3-4 days, selecting a strain with rapid color change and regular hyphae growth as a mother strain, then inoculating the mother strain into a seed culture medium, and performing culture for 5-7 days under the conditions that the temperature is 26-28 ℃ and the rotating speed of a shaking table is 150 plus materials at 200r/min to respectively obtain seed solutions of the edible fungi;
(2) ultrafine pulverizing herba Hyperici Japonici to 80-100 μm, adding 2-4 times of water, soaking at 30-40 deg.C for 1-2 days, and centrifuging to obtain herba Hyperici Japonici soaking extract;
(3) mixing the edible fungus seed liquids prepared in the step (1) according to an equal volume ratio in advance, inoculating 5-10% of inoculum size into a 5L fermentation tank of a high-density fermentation culture medium, adding the herba Hyperici Japonici soaking extract in the step (2) to start fermentation culture, wherein the mass ratio of the herba Hyperici Japonici soaking extract to the edible fungus seed liquids is 3-6: 1, the temperature is 28-30 ℃, the rotating speed is 100-250r/min, and the ventilation volume is 0.5-1.5dm3Fermenting and culturing for 5-10 days at min to obtain mixture of edible fungus fermented tea, centrifuging, and filtering supernatant;
(4) and (4) concentrating the filtrate obtained by filtering in the step (3) under reduced pressure at the temperature of below 60 ℃ until the water content is lower than 8%, and then freeze-drying to obtain powder, namely the camellia sinensis fermented product.
3. The method for preparing a camellia sinensis leaf fermented product using edible fungi according to claim 2, wherein the slant culture medium in the step (1) is a PDA culture medium or a PDPA complex culture medium.
4. The method for preparing a fermented product of herba Hyperici Japonici by using edible fungi as claimed in claim 2, wherein the seed culture medium in step (1) is: pH6.0-6.5, calculated by 100% of total mass fraction, composed of glucose 1.5-2.5%, peptone 0.1-0.2%, beef extract 0.1-0.2%, KH2PO4 0.1-0.15％、MgSO40.1-0.15%, vitamin B10.01-0.02%, and water in balance.
5. The method of claim 2The preparation method of the camellia sinensis leaf fermentation product by using edible fungi is characterized in that in the step (3), the pH of the fermentation medium is 5.5-6.0, and the fermentation medium comprises 2-3% of glucose, 0.4-0.6% of peptone, 0.2-0.4% of yeast extract and KH according to 100% of total mass fraction2PO4 0.15-0.3％、MgSO40.1-0.15%, vitamin B10.01-0.02%, and water in balance.
6. The use of the fermented product of camellia sinensis leaves of edible fungi as a moisturizer of skin care products as set forth in claim 1.
7. The use of the kohlrabi leaf fermented product using the edible fungi as the skin care product moisturizer as claimed in claim 1, wherein the mass percentage of the kohlrabi leaf fermented product using the edible fungi in the skin care product is 0.1-20%.
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| CN1891027A (en) * | 2002-07-18 | 2007-01-10 | 福建省农业科学院土壤肥料研究所 | Process for producing Grifolar Frondosus fermented tea |
| CN104872664A (en) * | 2015-05-26 | 2015-09-02 | 成都易创思生物科技有限公司 | Method for manufacturing health product by utilizing cordyceps grown in Nagqu Prefecture |
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