CN116270360A - Honeysuckle flower fermentation liquor and preparation method and application thereof - Google Patents
Honeysuckle flower fermentation liquor and preparation method and application thereof Download PDFInfo
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- CN116270360A CN116270360A CN202310398204.4A CN202310398204A CN116270360A CN 116270360 A CN116270360 A CN 116270360A CN 202310398204 A CN202310398204 A CN 202310398204A CN 116270360 A CN116270360 A CN 116270360A
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- honeysuckle
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- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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Abstract
The invention provides honeysuckle flower fermentation liquor and a preparation method and application thereof, and belongs to the technical field of biological fermentation. The invention ferments the honeysuckle homogeneous liquid by means of specific parameters and the secondary fermentation process of specific strains, and solves the problem of limited efficacy of the honeysuckle fermentation product in the prior art. The preparation method disclosed by the invention can be used for remarkably improving the activity of the honeysuckle in the aspect of cosmetic application, and especially improving the whitening and wrinkle removing performances of the honeysuckle. The tyrosinase inhibition rate of the honeysuckle flower fermentation liquor prepared by the method is up to 69.39%, the effects of removing wrinkles and smoothing skin are also obviously improved, and the honeysuckle flower fermentation liquor has the effects of relieving and resisting allergy of the honeysuckle flower and has good application prospects.
Description
Technical Field
The invention relates to the technical field of biological fermentation, in particular to honeysuckle flower fermentation liquor and a preparation method and application thereof.
Background
Honeysuckle flower, also called honeysuckle flower, is a flower of honeysuckle (Lonicera japonica Thunb.) belonging to the family Caprifoliaceae, which is initially white and then yellow, so that the honeysuckle flower is an important medicinal and edible plant, and has the effects of clearing heat, detoxicating, resisting bacteria and viruses, resisting allergy, reducing blood sugar and lipid, resisting oxidization, resisting tumor and the like. The honeysuckle flower contains organic acid, flavone, polysaccharide and other components, has the functions of resisting aging, inhibiting bacteria, diminishing inflammation and the like, and has wide application prospect in the field of cosmetics.
The microbial fermentation extraction method is a process for extracting the plant active ingredients and performing bioconversion, and has the advantages of high yield of the active ingredients, various effects, avoidance of damage to the plant active ingredients, high bioavailability of the active ingredients and the like. The mixed liquor rich in active substances, which is produced by fermenting plant materials by microorganisms, is called plant fermentation extract (Plant Fermentation Extracts, PFEs), has the characteristics of antioxidation, antibiosis, whitening, anti-aging and the like, and is widely applied to the fields of cosmetics and health care products.
CN114668699a discloses a preparation method of honeysuckle fermentation primary pulp, which adopts bacillus subtilis to ferment honeysuckle, and the prepared honeysuckle fermentation primary pulp has active ingredients for removing DPPH free radicals and active ingredients for inhibiting tyrosinase activity and melanin synthesis. CN114948819a discloses a preparation method of a honeysuckle flower ferment with repairing effect, which comprises the steps of adding saccharomycetes and/or lactobacillus for fermentation after the honeysuckle flower is subjected to enzymolysis and sterilization, has free radical scavenging capability, has repairing effect, and can be used for repairing skin damage resistance. However, the improvement degree of the two fermentation technologies on the effect of the honeysuckle is limited, and other existing technologies for fermenting the honeysuckle by utilizing a microbial fermentation extraction method are also required to be improved.
Disclosure of Invention
The invention aims to provide honeysuckle flower fermentation liquor, a preparation method and application thereof, which are used for solving the problem of limited efficacy of honeysuckle flower fermentation products in the prior art.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of honeysuckle flower fermentation liquor, which comprises the following steps:
mixing honeysuckle and water, homogenizing to obtain a honeysuckle homogeneous solution;
mixing the honeysuckle homogeneous solution, the Bradyyeast solution and the lactobacillus plantarum solution, and performing primary fermentation to obtain a honeysuckle primary fermentation solution;
sterilizing the primary fermentation liquid of the honeysuckle, inoculating bifidobacterium adolescentis, and performing secondary fermentation to obtain the fermentation liquid of the honeysuckle.
Preferably, the weight ratio of the honeysuckle to the water is 1:0.8-1.2;
the homogenizing rotating speed is 4000-6000 rpm;
the homogenizing time is 5-15 min.
Preferably, the live bacteria concentration of the Saccharomyces boulardii liquid is 1.0X10 9 ～1.0×10 10 CFU/mL；
The viable bacteria concentration of the lactobacillus plantarum bacterial liquid is 1.0x10 9 ～1.0×10 10 CFU/mL。
Preferably, the volume ratio of the honeysuckle homogeneous liquid to the Bradyyeast liquid to the lactobacillus plantarum liquid is 200:3-5:1.
Preferably, the temperature of the first fermentation is 27.5-28.5 ℃;
the rotating speed of the first fermentation is 400-600 rpm;
the time of the first fermentation is 5-10 d.
Preferably, the sterilization adopts normal pressure intermittent sterilization, the normal pressure intermittent sterilization comprises alternately performed working phases and intermittent phases, the temperature of the working phases is 70-80 ℃, the time of the working phases is 50-70 min, the temperature of the intermittent phases is 35-37 ℃, and the time of the intermittent phases is 0.5-1.5 d.
Preferably, the inoculation amount of the bifidobacterium adolescentis is 40-60 g/L.
Preferably, the temperature of the second fermentation is 37-39 ℃;
the rotating speed of the second fermentation is 500-700 rpm;
the time of the second fermentation is 25-35 d.
The invention also provides the honeysuckle flower fermentation liquor obtained by the preparation method.
The invention also provides application of the honeysuckle flower fermentation liquor in preparing cosmetics, wherein the cosmetics are products for relieving, whitening and/or removing wrinkles.
The invention has the technical effects and advantages that:
according to the invention, a group of special strains are obtained through a large number of screening experiments, and the honeysuckle homogeneous liquid is fermented by combining secondary fermentation with specific parameters, so that the activity of the honeysuckle in the aspect of cosmetic application can be remarkably improved, and particularly the whitening and wrinkle removing activities of the honeysuckle can be improved. The tyrosinase inhibition rate of the honeysuckle flower fermentation liquor prepared by the method is up to 69.39%, the effects of removing wrinkles and smoothing skin are also remarkably improved, and the honeysuckle flower fermentation liquor also has the effects of relieving and resisting allergy of the honeysuckle flower and has good application prospects.
Detailed Description
The invention provides a preparation method of honeysuckle flower fermentation liquor, which comprises the following steps: mixing honeysuckle and water, homogenizing to obtain a honeysuckle homogeneous solution; mixing the honeysuckle homogeneous solution, the Bradyyeast solution and the lactobacillus plantarum solution, and performing primary fermentation to obtain a honeysuckle primary fermentation solution; sterilizing the primary fermentation liquid of the honeysuckle, inoculating bifidobacterium adolescentis, and performing secondary fermentation to obtain the fermentation liquid of the honeysuckle; in the invention, the honeysuckle is preferably mixed with water after the impurities are removed by cleaning, and the weight ratio of the honeysuckle to the water is preferably 1:0.8-1.2, and more preferably 1:0.9-1.1; the rotation speed of the homogenization is preferably 4000 to 6000rpm, more preferably 4500 to 5500rpm; by a means ofThe homogenizing time is preferably 5 to 15 minutes, more preferably 8 to 12 minutes; the live bacteria concentration of the Saccharomyces boulardii liquid is preferably 1.0X10 9 ～1.0×10 10 CFU/mL, more preferably 8.0X10 9 ～2.0×10 10 CFU/mL; the viable bacteria concentration of the lactobacillus plantarum bacterial liquid is preferably 1.0x10 9 ～1.0×10 10 CFU/mL, more preferably 7.0X10 9 ～3.0×10 10 CFU/mL. The volume ratio of the honeysuckle homogeneous solution, the Bradyyeast solution and the lactobacillus plantarum solution is preferably 200:3-5:1, and more preferably 200:3.5-4.5:1.
In the present invention, the temperature of the first fermentation is preferably 27.5 to 28.5 ℃, and more preferably 27.8 to 28.2; the rotation speed of the first fermentation is preferably 400-600 rpm, more preferably 450-550 rpm; the time of the first fermentation is preferably 5-10 d, more preferably 7-9 d, the sterilization is preferably an atmospheric intermittent sterilization, the atmospheric intermittent sterilization comprises alternating working phases and intermittent phases, the temperature of the working phases is preferably 70-80 ℃, more preferably 73-77 ℃, the time of the working phases is preferably 50-70 min, more preferably 55-65 min, the temperature of the intermittent phases is preferably 35-37 ℃, more preferably 35.5-36.5 ℃, the time of the intermittent phases is preferably 0.5-1.5 d, more preferably 0.8-1.2 d; the inoculation amount of the bifidobacterium adolescentis is preferably 40-60 g/L, more preferably 45-55 g/L, and the temperature of the second fermentation is preferably 37-39 ℃, more preferably 37.5-38.5 ℃; the rotation speed of the second fermentation is preferably 500-700 rpm, more preferably 550-650 rpm; the time for the second fermentation is preferably 25 to 35d, more preferably 28 to 32d.
The invention also provides the honeysuckle flower fermentation liquor obtained by the preparation method, the fermentation liquor is preferably light yellow in state, the smell is preferably light grass and wood fragrance, no peculiar smell is generated, and meanwhile, the honeysuckle flower fermentation liquor is taken as a sign of successful fermentation.
The invention also provides application of the honeysuckle fermentation liquor in preparing cosmetics, the honeysuckle fermentation liquor is preferably used for preparing whitening and/or wrinkle removing products, the formulation of the whitening and/or wrinkle removing products is preferably cream, mask, emulsion, toning lotion, bath foam, facial cleanser and the like, the addition amount of the honeysuckle fermentation liquor in the products is preferably 5-100%, and more preferably 40-90%, and in the invention, the whitening and/or wrinkle removing products also comprise auxiliary materials acceptable by cosmetics, and the auxiliary materials comprise moisturizers, emulsifiers, thickeners, skin conditioning agents, pH regulators, preservatives, water and the like; the emulsifier is preferably sodium lauroyl glutamate, lauramidopropyl betaine, isopropyl myristate or cocoyl glucoside; the humectant is preferably sodium hyaluronate, glycerol, propylene glycol, butylene glycol, dipropylene glycol, sorbitol, inositol, beta-glucan or trehalose; the thickener is preferably disodium EDTA, carbomer, xanthan gum, ammonium acryloyldimethyl taurate/VP copolymer or sodium polyacrylate grafted starch; the skin conditioning agent is preferably octanoyl hydroxamic acid, allantoin, ceramide, nicotinamide, vitamin C derivatives, arbutin, tranexamic acid, yeast/hydrolytic yeast; the pH regulator is preferably arginine, sodium citrate, citric acid, phosphoric acid, tartaric acid, sodium dihydrogen phosphate and triethanolamine, the preservative is preferably methylparaben, butylparaben, ethylparaben, isobutyl paraben, propyl paraben, potassium sorbate and sodium benzoate, and the auxiliary materials preferably do not influence the efficacy of the honeysuckle fermentation broth.
The technical effects provided by the present invention will be described in detail with reference to examples, but they should not be construed as limiting the scope of the present invention.
Lactobacillus plantarum lyophilized powder (lactobacillus plantarum) SHBCCD 81363, saccharomyces boulardii lyophilized powder (Saccharomyces cerevisiae var boulardii) CNCM I-1079, candida mycota (Candida boleticola) SHBCC D53761, lactobacillus rhamnosus lyophilized powder (Lactobacillus rhamnosus) SHBCC D80286, purchased from Shanghai preservation biotechnology center;
bifidobacterium adolescentis probiotic freeze-dried powder (product number: ZKJY-Bifidobacterium adolescentis) is purchased from the micro-ecological research institute of Carnis nutritional medicine (Shandong) of the middle-jiao, and the viable count is more than or equal to 1.0X10 10 CFU/g；
Lactobacillus paracasei freeze-dried lactobacillus powder (product number: LP 075), and viable count is not less than 1.0X10 10 CFU/g, lactobacillus bulgaricus freeze-dried lactobacillus powder (product number: LB 005), viable count is not less than 1.0X10 10 CFU/g, available from Zhengzhou Bai Bao Biotechnology Co.
Example 1
Putting 1kg of cleaned and impurity-removed honeysuckle flower into a container, adding 1kg of water, homogenizing for 10min by using a homogenizer (SL-RH-LAB 200 homogenizing emulsifying machine) at a rotation speed of 5000rpm to obtain a honeysuckle flower homogenized liquid for later use.
Activating the freeze-dried powder of the Buddy yeast according to the commodity instruction, then passaging twice, and performing expanded culture to 5.0X10 by adopting malt extract broth culture medium without adding agar 9 CFU/mL, lactobacillus plantarum freeze-dried powder is activated according to a commodity instruction and then passaged twice, and the agar-free MRS culture medium is subjected to expansion culture until the concentration is 5.0x10 9 CFU/mL, inoculating 20mL of Bradyyeast liquid and 5mL of lactobacillus plantarum liquid into 1L of honeysuckle homogeneous liquid, transferring a fermentation system into a stainless steel fermentation tank, and fermenting for 7d at 28 ℃ and 500rpm to obtain a honeysuckle primary fermentation liquid.
Intermittently sterilizing the primary fermentation broth of flos Lonicerae, heating at 75deg.C for 60min, cooling to 37deg.C, culturing for 1d, heating at 75deg.C for 60min, and cooling to 37deg.C, and culturing for 12 h.
Inoculating the bifidobacterium adolescentis probiotic freeze-dried powder into the sterilized honeysuckle primary fermentation broth with the inoculation amount of 50g/L, transferring the honeysuckle primary fermentation broth into a stainless steel fermentation tank, and fermenting for 30d at 38 ℃ and 600rpm to obtain a honeysuckle fermentation broth final product.
Example 2
1kg of cleaned and impurity-removed honeysuckle flower is put into a container, 0.8kg of water is added, and the mixture is homogenized for 5min under the condition of 4000rpm of a homogenizer (SL-RH-LAB 200 homogenizing emulsifying machine) to obtain a honeysuckle flower homogenized liquid for standby.
Activating the freeze-dried powder of the Buddy yeast according to the commodity instruction, then passaging twice, and performing expanded culture to 1.0X10 by adopting malt extract broth culture medium without adding agar 9 CFU/mL, plantsThe lactobacillus freeze-dried powder is activated according to the commodity instruction and then passaged twice, and the MRS culture medium without agar is subjected to expansion culture until the concentration is 1.0x10 10 CFU/mL, inoculating 25mL of Bradyyeast liquid and 5mL of lactobacillus plantarum liquid into 1L of honeysuckle homogeneous liquid, transferring a fermentation system into a stainless steel fermentation tank, and fermenting for 5d at 28.5 ℃ and 400rpm to obtain a honeysuckle primary fermentation liquid.
Intermittently sterilizing the primary fermentation broth of flos Lonicerae, heating at 70deg.C for 50min, cooling to 36deg.C, culturing for 1.5d, heating at 75deg.C for 70min again, cooling to 37deg.C, culturing for 0.5d, heating at 75deg.C for 50min again, and cooling to 36deg.C, and culturing for 10 hr.
Inoculating the bifidobacterium adolescentis probiotic freeze-dried powder into the sterilized honeysuckle primary fermentation broth with the inoculation amount of 40g/L, transferring the honeysuckle primary fermentation broth into a stainless steel fermentation tank, and fermenting for 25d at 39 ℃ and 500rpm to obtain a honeysuckle fermentation broth final product.
Example 3
1kg of cleaned and impurity-removed honeysuckle flower is put into a container, 1.2kg of water is added, and the mixture is homogenized for 15min by a homogenizer (SL-RH-LAB 200 homogenizing emulsifying machine) at a rotating speed of 6000rpm, so as to obtain a honeysuckle flower homogenized liquid for standby.
Activating the freeze-dried powder of the Buddy yeast according to the commodity instruction, then passaging twice, and performing expanded culture to 1.0X10 by adopting malt extract broth culture medium without adding agar 10 CFU/mL, lactobacillus plantarum freeze-dried powder is activated according to a commodity instruction and then passaged twice, and the agar-free MRS culture medium is subjected to expansion culture until the concentration is 1.0x10 9 CFU/mL, inoculating 15mL of Bradyyeast liquid and 5mL of lactobacillus plantarum liquid into 1L of honeysuckle homogeneous liquid, transferring a fermentation system into a stainless steel fermentation tank, and fermenting for 10d at the temperature of 27.5 ℃ and the speed of 600rpm to obtain a honeysuckle primary fermentation liquid.
Intermittently sterilizing the primary fermentation broth of flos Lonicerae, heating at 80deg.C for 70min, cooling to 35deg.C, culturing for 0.5d, heating again at 75deg.C for 70min, cooling to 35deg.C, culturing again for 1.5d, heating again at 75deg.C for 50min, and cooling to 35deg.C, and culturing for 14 h.
Inoculating the bifidobacterium adolescentis probiotic freeze-dried powder into the sterilized honeysuckle primary fermentation broth with the inoculation amount of 60g/L, transferring the honeysuckle primary fermentation broth into a stainless steel fermentation tank, and fermenting for 35d at 37 ℃ and 700rpm to obtain a honeysuckle fermentation broth final product.
Comparative example 1
The only difference compared to example 1 is that the brainstem yeast lyophilized powder was replaced with candida mycota.
Comparative example 2
The only difference compared to example 1 is that the lactobacillus plantarum freeze-dried powder is replaced by lactobacillus rhamnosus freeze-dried powder.
Comparative example 3
The only difference compared to example 1 is that the bifidobacterium adolescentis probiotic freeze-dried powder is replaced by lactobacillus paracasei freeze-dried lactic acid bacteria powder.
Comparative example 4
The only difference compared to example 1 is that the freeze-dried powder of bifidobacterium adolescentis probiotics is replaced by the freeze-dried powder of lactobacillus bulgaricus.
Experimental example 1 verification of whitening Effect
The sample sources are shown in table 1 below:
TABLE 1 Experimental sample Source
| Experimental grouping | Sample source |
| Experimental group A | Honeysuckle homogeneous solution in example 1 |
| Experiment group B | Honeysuckle primary fermentation liquor in example 1 |
| Experiment group C | The honeysuckle flower fermentation broth end product in example 1 |
| Experiment group D | The final product of comparative example 1 |
| Experiment group E | The final product of comparative example 2 |
| Experiment group F | The final product of comparative example 3 |
| Experiment group G | The final product of comparative example 4 |
The whitening efficacy of each group of samples was measured separately.
Referring to the method of Aoki Y et al (Aoki Y, tanigawa T, abe H, et al, melanogenesis inhibi-tion by an oolong tea extract in B mouse melanoma cells and UV-induced skin pigmentation in brownish guinea pigs [ J ]. Biosci Biotechnol Biochem,2007,71 (8): 1879-1885.), each group of samples was characterized by tyrosinase inhibition, and the positive control was assayed in triplicate using a 50 μg/mL concentration of kojic acid, and the tyrosinase inhibition (%) was recorded for each group, as shown in Table 2 below:
TABLE 2 tyrosinase inhibition results
The results show that the final product of the honeysuckle fermentation liquor in the embodiment 1 shows the highest tyrosinase inhibition rate, which shows that the whitening effect is optimal, compared with the honeysuckle raw material, the effect is obviously improved, and the effect is superior to other strain combinations.
Experimental example 2
The sample sources were the same as in experimental example 1, and the samples were pasteurized for 30min and then used for the test.
Selecting 35 female volunteers with eye marks, wherein the exclusion conditions meet the exclusion standard of cosmetic contact dermatitis diagnostic standard and treatment principle, the conditions of non-allergic physique or skin diseases exist, and the women in non-pregnant women or lactation period do not participate in other cosmetic experiments 2 months before the experiment; the same experimental group samples were used randomly for every 5 volunteers.
Wrinkle removal performance of each group of samples was tested using a VisioScan VC20 plus skin surface texture tester, and R3 and S were recorded esm The lower these two indices, the smoother the skin, the lower the visibility of wrinkles; the subjects respectively use the same cleaning product to clean the face in the same environment (the temperature is 27 ℃ and the test is started at four afternoon), after sitting still for 30min, the initial values are measured, then the samples are respectively smeared at the eye marks, the samples are massaged to be absorbed, and after 30min, the test is performed again; the test was performed at four afternoon points 7d, 14d, 28d, respectively, using the same procedure each day, and the results were recorded as shown in table 3 below:
table 3 texture test results
The results show that the two-step fermentation method provided by the invention is matched with specific strains, so that the wrinkle removing effect of the honeysuckle fermentation liquid can be improved, the skin is smoother, and the honeysuckle fermentation product provided by the invention can be improved in the effect of delaying aging and has a good application prospect.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. The preparation method of the honeysuckle flower fermentation liquor is characterized by comprising the following steps of:
mixing honeysuckle and water, homogenizing to obtain a honeysuckle homogeneous solution;
mixing the honeysuckle homogeneous solution, the Bradyyeast solution and the lactobacillus plantarum solution, and performing primary fermentation to obtain a honeysuckle primary fermentation solution;
sterilizing the primary fermentation liquid of the honeysuckle, inoculating bifidobacterium adolescentis, and performing secondary fermentation to obtain the fermentation liquid of the honeysuckle.
2. The preparation method of the honeysuckle fermentation broth according to claim 1, wherein the weight ratio of the honeysuckle to the water is 1:0.8-1.2;
the homogenizing rotating speed is 4000-6000 rpm;
the homogenizing time is 5-15 min.
3. The method for preparing the honeysuckle fermentation broth according to claim 2, wherein the live bacteria concentration of the brazil yeast liquid is 1.0 x 10 9 ～1.0×10 10 CFU/mL；
The viable bacteria concentration of the lactobacillus plantarum bacterial liquid is 1.0x10 9 ～1.0×10 10 CFU/mL。
4. The preparation method of the honeysuckle fermentation broth according to claim 3, wherein the volume ratio of the honeysuckle homogeneous solution, the branchia yeast solution and the lactobacillus plantarum solution is 200:3-5:1.
5. The method for preparing honeysuckle fermentation broth according to claim 4, wherein the temperature of the first fermentation is 27.5-28.5 ℃;
the rotating speed of the first fermentation is 400-600 rpm;
the time of the first fermentation is 5-10 d.
6. The preparation method of honeysuckle flower fermentation broth according to claim 5, wherein the sterilization adopts normal pressure intermittent sterilization, the normal pressure intermittent sterilization comprises alternately performed working phases and intermittent phases, the temperature of the working phases is 70-80 ℃, the time of the working phases is 50-70 min, the temperature of the intermittent phases is 35-37 ℃, and the time of the intermittent phases is 0.5-1.5 d.
7. The method for preparing a honeysuckle fermentation broth according to claim 6, wherein the inoculation amount of bifidobacterium adolescentis is 40-60 g/L.
8. The method for preparing honeysuckle fermentation broth according to claim 7, wherein the temperature of the second fermentation is 37-39 ℃;
the rotating speed of the second fermentation is 500-700 rpm;
the time of the second fermentation is 25-35 d.
9. The honeysuckle flower fermentation broth obtained by the preparation method of any one of claims 1 to 8.
10. Use of the honeysuckle fermentation broth according to claim 9 for the preparation of a cosmetic product, characterized in that the cosmetic product is a soothing, whitening and/or wrinkle-removing product.
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