CN113318037A - Microbial fermentation method for increasing content of active ingredients in peony and application - Google Patents

Microbial fermentation method for increasing content of active ingredients in peony and application Download PDF

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CN113318037A
CN113318037A CN202110776090.3A CN202110776090A CN113318037A CN 113318037 A CN113318037 A CN 113318037A CN 202110776090 A CN202110776090 A CN 202110776090A CN 113318037 A CN113318037 A CN 113318037A
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peony
fermentation
content
active ingredients
fermentation broth
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CN113318037B (en
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刘向慧
张岩松
张丽雷
陈叶
谢婉月
杨会枝
押辉远
刘献明
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Luoyang Normal University
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • 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
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    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K2800/782—Enzyme inhibitors; Enzyme antagonists
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    • 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
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Abstract

The invention relates to the field of plant application methods, and particularly discloses a microbial fermentation method for improving the content of active ingredients of peony, which comprises the following steps: s1, drying and crushing fresh peony into peony powder, mixing with distilled water, and performing ultrasonic treatment to obtain a mixed solution for later use; s2, inoculating microorganisms into the mixed liquor obtained in S1, then performing fermentation culture, sterilizing, centrifuging and taking supernatant to obtain peony fermentation liquor, wherein the fermentation conditions are as follows: the inoculation amount is 1-10%, the temperature is 36-37 ℃, the fermentation time is 12-48h, and the initial pH is 5-7. The content of peony flavone and polyphenol in the peony fermentation liquid prepared by the fermentation method is obviously improved, and the antioxidant and whitening effects are also obviously improved.

Description

Microbial fermentation method for increasing content of active ingredients in peony and application
Technical Field
The invention relates to the field of plant application methods, in particular to a microbial fermentation method for improving the content of active ingredients of peony and application thereof.
Background
Peony (Paeonia suffruticosa Andr.) is a plant of the genus Paeonia, the family Paeoniaceae, and the class Dicotyledoneae, and is a perennial deciduous shrub. It is mainly produced in the fields of Shandong lotus and Henan Luo Yang. It is one of the most important ornamental plants in the international flower market. The flower is large and single-grown, usually dark red, pink and white, is popular with people because the flower is rich and luxurious, beautiful and modesty, and is easily reputed by the king in the flower. The peony has not only ornamental value, but also important economic value. The peony flowers published by the national institutes of health in 10 months in 2013 can be used as a new food raw material, and the peony series extracts are republished by the national drug and food supervision and management administration in 6 months in 2015 to be used as cosmetic raw materials. The peony contains rich nutrients necessary for organisms, including vitamins, proteins, saccharides, anthocyanins, mineral elements and the like, and has the functions of resisting aging and resisting external adverse environment. In addition, the peony also contains flavones such as astragalin, gallic acid and the like and polyphenol compounds, and the substances not only can remove free radicals in the chain reaction initiation stage, but also can directly capture free radicals in the free radical reaction chain, thereby playing a dual anti-oxidation role in blocking the free radical reaction chain. Researchers analyze the whole phenolic section and the antioxidant activity of different organs of the peony (red phoenix white), and the content of flavonoid glycoside in the petals is higher than that of other organs, so that the peony is a potential functional food raw material and a natural antioxidant raw material.
Peony is used as a cosmetic raw material and has the effects of resisting oxidation and whitening skin, but the capacity of obtaining effective components by a common extraction method is limited, and the problem of solubility exists, so that the application of the peony extract in cosmetics is limited and the peony extract is not popularized.
Disclosure of Invention
In order to solve the technical problems, the invention provides a microbial fermentation method for increasing the content of active ingredients of peony and application thereof.
The invention aims to provide a microbial fermentation method for improving the content of active ingredients in peony, which comprises the following steps:
s1, drying and crushing fresh peony into peony powder, mixing with distilled water, and performing ultrasonic treatment to obtain a mixed solution for later use;
s2, sterilizing and cooling the mixed solution prepared in the S1, inoculating probiotics, performing fermentation culture, centrifuging, and taking supernatant to obtain peony fermentation liquor;
the probiotics are lactobacillus plantarum or streptococcus thermophilus;
the fermentation conditions were: the inoculation amount is 1-10 vol%, the temperature is 36-37 ℃, the fermentation time is 12-48h, and the initial pH of the fermentation liquid is 5-7.
Preferably, in S2, the fermentation conditions are: the inoculation amount is 1 vol%, the fermentation time is 36h, and the initial pH of the fermentation liquid is 6.5.
Preferably, in S1, the ratio of the peony powder to distilled water is 7.5 g: 140 and 160 mL.
Preferably, in S1, the drying condition is vacuum drying at 50 ℃.
Preferably, in S1, the ultrasonic treatment condition is 60 ℃ for 30 min.
Preferably, in S2, the centrifugation is performed for 10min under 10000 r/min.
The second purpose of the invention is to provide a peony fermentation liquid prepared by the microbial fermentation method.
The third purpose of the invention is to provide the application of the peony fermentation liquor in preparing skin care products.
Preferably, the peony fermentation liquor is used for preparing an antioxidant skin care product.
Preferably, the peony fermentation liquor is used for preparing a skin care product with a whitening effect.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the microbial fermentation method, the peony fermentation broth is obtained through fermentation under specific fermentation conditions (the inoculation amount is 1-10 vol%, the temperature is 36-37 ℃, the fermentation time is 12-48h, and the initial pH is 5-7), detection results show that the content of peony flavone and polyphenol in the peony fermentation broth is obviously improved, and the determination of DPPH free radical, hydroxyl free radical, ABTS free radical clearance rate and tyrosinase inhibition rate shows that the peony fermentation broth obtained through the method provided by the invention has the advantages that the oxidation resistance and whitening effect are obviously improved, and the application prospect for preparing skin care products is realized.
2. The invention finds that the optimum setting inoculation amount is 1 vol%, the temperature is 36 ℃, the fermentation time is 36h, and the initial pH is 6.5, the peony fermentation broth has the highest content promotion degree on peony flavone and polyphenol, and the antioxidation effect is also the most obvious (and has obvious difference with other parameters).
3. The method combines the traditional plant resources of China with the modern fermentation technology, utilizes peony and probiotics to ferment to obtain peony fermentation liquor, and the peony fermentation liquor has advantages in the aspects of safety, toxic and side effects, effectiveness and the like and has great application potential in the field of cosmetic application.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a graph showing the growth of Lactobacillus plantarum in accordance with the present invention;
FIG. 2 shows the differences of Lactobacillus plantarum in the present inventionIC of flavone content and DPPH free radical in peony fermentation liquor under inoculation amount condition50A value;
FIG. 3 shows IC of flavone content and DPPH free radical in peony fermentation broth at different fermentation times in the present invention50A value;
FIG. 4 shows IC of xanthone content and DPPH radical in peony fermentation broth at different initial pH values in the present invention50A value;
FIG. 5 shows the polyphenol content in the water extract of peony, the water extract of sterilized peony, the fermentation broth of Streptococcus thermophilus of peony and the fermentation broth of Lactobacillus plantarum of peony in the present invention;
FIG. 6 shows the xanthone content in the water extract of peony, the water extract of sterilized peony, the fermentation broth of Streptococcus thermophilus of peony and the fermentation broth of Lactobacillus plantarum of peony in the present invention;
FIG. 7 shows the scavenging rates of the peony water extract, the sterilized peony water extract, the peony streptococcus thermophilus fermentation broth and the peony lactobacillus plantarum fermentation broth DPPH free radical in the present invention;
FIG. 8 shows the hydroxyl radical scavenging rate of the peony water extract, the sterilized peony water extract, the peony streptococcus thermophilus fermentation broth and the peony lactobacillus plantarum fermentation broth in the present invention;
FIG. 9 shows the radical scavenging rates of the peony water extract, the sterilized peony water extract, the peony streptococcus thermophilus fermentation broth and the peony lactobacillus plantarum fermentation broth ABTS in the present invention.
FIG. 10 shows the tyrosinase inhibition rates of the peony water extract, the sterilized peony water extract, the peony streptococcus thermophilus fermentation broth, and the peony lactobacillus fermentation broth of the present invention.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. The experimental methods described in the examples of the present invention are all conventional methods unless otherwise specified.
Example 1
The embodiment provides a microbial fermentation method for improving the content of active ingredients of peony.
Lactobacillus plantarum fermented peony
1. Determination of strain growth curve
Inoculating activated Lactobacillus plantarum into MRS liquid culture medium, culturing in a constant temperature incubator at 36 deg.C, determining viable count at corresponding time by plate colony counting method, taking out at corresponding time, transferring into a cuvette, determining absorbance of Lactobacillus plantarum culture solution at 600nm with distilled water as blank control, recording, taking culture time as abscissa, and viable count and OD600A growth curve was plotted on the ordinate (as shown in FIG. 1), and the optimal activation culture time of Lactobacillus plantarum was determined to be 24 h.
2. Raw material treatment
Drying fresh peony in a vacuum drying oven at 50 ℃, crushing dried peony into peony powder, sieving with a 80-mesh sieve, weighing 7.5g, mixing with 150mL of distilled water, performing ultrasonic treatment at 60 ℃ for 30min, centrifuging at 10000r/min for 10min, and taking supernatant, namely peony water extract, as a control. Sterilizing with high pressure steam at 121 deg.C for 20min after ultrasonic treatment, centrifuging under the same conditions for 10min, and collecting supernatant to obtain sterilized water extractive solution of flos moutan.
3. Microbial fermentation
Fresh peony is dried in a vacuum drying oven at 50 ℃, the dried peony is made into peony powder by a pulverizer, the peony powder is sieved by a 80-mesh sieve, 7.5g of the peony powder is weighed and mixed with 150mL of distilled water, the mixture is subjected to ultrasonic treatment at 60 ℃ for 30min and then is sterilized by high-pressure steam at 121 ℃ for 20min, lactobacillus plantarum activated for 24h is added into the mixture according to the inoculation amount of 1 vol% after the mixture is cooled to room temperature, the mixture is cultured in a constant-temperature incubator at 36 ℃ for 48h, the culture medium is centrifuged (10000rpm and 10min) after 48h to obtain peony fermentation liquor, and the peony water extract is used as a reference to respectively measure the flavone content and DPPH removing capacity of the peony water extract, the sterilized peony water and the peony fermentation liquor.
Example 2
This example is substantially the same as example 1, except that:
the inoculation amount of lactobacillus plantarum was 2 vol%.
Example 3
This example is substantially the same as example 1, except that:
the inoculation amount of lactobacillus plantarum was 5 vol%.
Example 4
This example is substantially the same as example 1, except that:
the inoculation amount of lactobacillus plantarum was 10 vol%.
The results of measuring the flavone content and DPPH removing ability of the peony fermentation broth obtained in the above examples 1 to 4 are shown in FIG. 2.
As shown in fig. 2, compared with the peony water extract, the peony fermentation broth prepared in examples 1 to 3 has significantly improved flavone content and DPPH scavenging ability, and has significant difference (# # P is less than or equal to 0.01);
compared with the sterilized water extract of peony, the peony fermentation broth prepared in example 1 has significant difference in flavone content (P is less than or equal to 0.01); there was a significant difference in DPPH clearance capacity (. about.0.01 < p.ltoreq.0.05).
Example 5
This example is substantially the same as example 1, except that:
culturing in 36 deg.C constant temperature incubator for 12h, respectively, i.e. fermentation time is 12 h.
Example 6
This example is substantially the same as example 1, except that:
respectively culturing in a constant temperature incubator at 36 ℃ for 24h, namely, the fermentation time is 24 h.
Example 7
This example is substantially the same as example 1, except that:
respectively culturing in a constant temperature incubator at 36 ℃ for 36h, namely, the fermentation time is 36 h.
The flavone content and DPPH removing ability of the peony fermentation broth obtained in the above examples 5 to 7 were measured, and the best effect was found in example 7. (as shown in FIG. 3), there was a significant difference (# # P ≦ 0.01) compared to the aqueous extract from peony, and a significant difference (# # P ≦ 0.01,. 0.01< P ≦ 0.05) compared to the aqueous extract from sterilized peony.
Example 8
This example is substantially the same as example 1, except that:
respectively adjusting the initial pH value of the mixed solution to 5 by using 1% NaOH solution, sterilizing the mixed solution for 20min by using high-pressure steam at the temperature of 121 ℃, cooling the mixed solution to room temperature, inoculating activated lactobacillus plantarum according to the proportion of 1 vol%, and using peony water extract and sterilized peony water extract as a control. Culturing in a constant temperature incubator at 36 deg.C for 24h, centrifuging (10000rpm, 10min), and collecting supernatant to obtain peony fermentation broth. The content of flavone in the water extract of peony, the sterilized water extract of peony and the fermentation liquid of peony and the DPPH removing ability are respectively measured.
Example 9
This example is substantially the same as example 8 except that:
the initial pH was adjusted to 6.
Example 10
This example is substantially the same as example 8 except that:
the initial pH was adjusted to 6.5.
Example 11
This example is substantially the same as example 8 except that:
the initial pH was adjusted to 7.
The xanthone contents and their DPPH removing abilities were measured in the water extracts of peony, the sterilized water extracts of peony and the fermentation solutions of peony of examples 8 to 11, respectively, and it was found that example 10 is the most preferable (as shown in FIG. 4), and that there is a significant difference (# # P ≦ 0.01) compared to the water extracts of peony and a significant difference (# # P ≦ 0.01; < P ≦ 0.05) compared to the water extracts of sterilized peony.
Example 12
Method for measuring content of flavone
(1) Preparation of a standard curve: accurately weighing 25mg of rutin standard substance, and diluting to 25mL with anhydrous ethanol to obtain rutin standard substance solution. Accurately sucking the reagent amount in Table 2, shaking, standing for 15min, and measuring absorbance at 510nm wavelength. And (3) drawing a standard curve by taking different rutin concentrations as a horizontal coordinate and the corresponding absorbance as a vertical coordinate, and solving a regression equation.
TABLE 1 rutin Standard Curve sample addition
Figure BDA0003154862180000081
And (3) total flavone content determination: and (3) carrying out reaction on the sample according to the method for preparing the standard curve to determine the absorbance value, substituting the absorbance value into a linear equation, and calculating the content of the total flavone.
(2) DPPH radical scavenging Rate determination
Loading samples according to the following table 2, placing the samples in dark for 30min, measuring the absorbance of the samples at the wavelength of 517nm, calculating the DPPH free radical clearance of different samples, and taking Vc as a positive control, wherein the DPPH free radical clearance is measured according to a formula:
Figure BDA0003154862180000082
in the formula, A0: absorbance values of blank control; and A, absorbance value of each sample.
TABLE 2 sample addition for DPPH free radical scavenging experiments
Figure BDA0003154862180000083
Secondly, probiotic fermented peony
1. Preparation of samples
Preparing peony powder from dried peony flowers by using a pulverizer, sieving with a 80-mesh sieve, weighing 7.5g of sieved pollen, mixing with 150mL of distilled water, performing ultrasonic treatment at 60 ℃ for 30min, centrifuging at 10000r/min for 10min, and taking supernatant fluid, namely the peony flower water extract. And (3) carrying out high-temperature high-pressure sterilization after the same ultrasonic treatment, cooling, centrifuging and taking supernatant to obtain the sterilized peony water extract.
Inoculating lactobacillus plantarum into an MRS culture medium for strain activation and propagation for 24h, inoculating 1 vol% of bacterial liquid into the non-centrifuged mixed liquid after high-temperature and high-pressure sterilization, and culturing in a constant temperature box at 37 ℃ for 24h to obtain a fermentation product. Centrifuging for 10min under 10000r/min, discarding the precipitate, and collecting the supernatant to obtain the peony plant lactobacillus fermentation liquid.
Inoculating the preserved streptococcus thermophilus into an M17 culture medium for strain propagation, inoculating 5 vol% of bacterial liquid into the non-centrifuged mixed liquid after high-temperature and high-pressure sterilization, and fermenting in a thermostat at 42 ℃ for 48 hours to obtain a fermentation product. And (4) performing the same post-treatment on the fermentation product as the lactobacillus plantarum fermentation liquor to obtain the peony streptococcus thermophilus fermentation liquor.
2. Change of main components before and after probiotic fermentation of peony
(1) And (3) measuring the polyphenol content: samples were loaded as in Table 3, left for 30min in the dark, absorbance was measured at 750nm, and 3 replicates were repeated. And (3) drawing a standard curve by taking different gallic acid concentrations as abscissa and the corresponding absorbance as ordinate, and solving a regression equation.
TABLE 3 Gallic acid Standard Curve for each solution addition
Figure BDA0003154862180000091
Figure BDA0003154862180000101
And (3) taking the peony water extract, the sterilized peony water extract, the peony streptococcus thermophilus fermentation broth and the peony plant lactobacillus fermentation broth, determining the polyphenol content of the samples according to the method for preparing the standard curve, recording the absorbance value of each sample, substituting into a linear equation, and calculating the polyphenol content.
(2) Measurement of flavone content
And (3) accurately weighing 25mg of rutin standard substance in the preparation of the standard curve, and fixing the volume to 25mL by using absolute ethyl alcohol to obtain a rutin standard substance solution. Accurately sucking the reagent amount in Table 2, shaking, standing for 15min, and measuring absorbance at 510nm wavelength. And (3) drawing a standard curve by taking different rutin concentrations as a horizontal coordinate and the corresponding absorbance as a vertical coordinate, and solving a regression equation.
And (3) taking the peony water extract, the sterilized peony water extract, the peony streptococcus thermophilus fermentation broth and the peony plant lactobacillus fermentation broth, determining the total flavone content of the samples according to the method for preparing the standard curve, recording the absorbance value of each sample, substituting into a linear equation, and calculating the total flavone content.
3. Before and after probiotic fermentation of peony, change of antioxidant and whitening effects
(1) DPPH radical scavenging Rate determination
DPPH free radical scavenging experiment has the advantages of simplicity, convenience, direct feasibility and is widely used for the research of antioxidants. DPPH + H+→ DPPH, which has a strong absorption at 517nm, the absorbance of the system at the maximum absorption wavelength becomes small and the solution color becomes light after the addition of the antioxidant substance.
Taking the reagents in the table 2, mixing uniformly, standing for 30min in the dark at room temperature, measuring the absorbance value at 517nm, and determining the DPPH free radical clearance formula:
Figure BDA0003154862180000102
in the formula, A0: absorbance values of blank control; and A, absorbance value of each sample.
(2) Measurement of hydroxyl radical scavenging Rate
The reaction mechanism of the hydroxyl radical scavenging test is: h2O2+Fe2+＝·OH+H2O+Fe3+Then salicylic acid is added into the reaction system, as shown in the following reaction formula, if a substance with the function of removing hydroxyl radicals is added into the reaction system, hydroxyl radicals are consumed, so that the hydroxyl radicals in the system are reduced, the generation of 2, 3-dihydroxy benzoic acid is reduced, namely, the generation amount of a colored compound is correspondingly reduced, and the absorbance is reduced when the reaction solution containing the antioxidant substance is measured at 510 nm.
Respectively taking the reagents in the table 4, mixing uniformly, standing at 37 ℃ for 30min, measuring the absorbance value by using an enzyme-labeling instrument at the wavelength of 510nm, and determining the hydroxyl radical clearance by using a formula:
Figure BDA0003154862180000111
TABLE 4 measurement of hydroxyl radical scavenging Rate
Figure BDA0003154862180000112
(3) ABTS free radical clearance assay
The reaction principle of ABTS free radical scavenging is as follows: ABTS + K2S2O8→ABTS+(blue-green) + K2SO4The ABTS produced by the reaction has maximum absorption peaks at 734nm, 414nm, 645nm and 805nm, and if a compound for scavenging free radicals reacts with the ABTS, the ABTS becomes colorless. The absorbance of the reaction was measured at 734nm and the corresponding clearance was calculated.
The reagents in Table 5 were mixed well, left in the dark at room temperature for 30min, and the absorbance value was measured at 734 nm. ABTS radical clearance was calculated for each sample according to equation (3).
Figure BDA0003154862180000113
In the formula, A0: absorbance values of blank control; and A, absorbance value of each sample.
TABLE 5 ABTS radical scavenging Rate determination test sample addition
Figure BDA0003154862180000114
(4) Determination of tyrosinase inhibitory Effect
The reagents in Table 6 were mixed well, left at 37 ℃ for 25min, and the absorbance was measured at 490 nm.
ABTS radical clearance was calculated for each sample according to equation (4).
Figure BDA0003154862180000121
In the formula, T is the absorbance of the enzyme mixed solution of the extract without adding the sample; t0 is the absorbance of the enzyme-free mixture of the extract without sample; c is the absorbance of the enzyme-added mixed solution of the sample-added extract; c0 is the absorbance of the enzyme mixture without the addition of the sample extract.
TABLE 6 tyrosinase inhibition experiment dosing sequence
Figure BDA0003154862180000122
4. The result of the detection
(1) The method is used for detecting the contents of polyphenol and flavone in the peony water extract, the sterile peony water extract, the peony streptococcus thermophilus fermentation liquor and the peony plant lactobacillus fermentation liquor, and the detection results are shown in fig. 5 and fig. 6.
The microbial fermentation method adopted by the invention obviously improves the contents of peony flavone and polyphenol.
(2) DPPH free radical clearance determination test results
The determination results of the DPPH free radical clearance after different gradient dilution of the peony water extract, the sterile peony water extract, the peony streptococcus thermophilus fermentation broth and the peony lactobacillus plantarum fermentation broth are shown in FIG. 7, with the decrease of the concentration, the DPPH clearance of the extract and the fermentation broth show the trend of increasing first and then decreasing, and the DPPH clearance of the peony streptococcus thermophilus fermentation broth and the lactobacillus plantarum fermentation broth is higher than that of the peony water extract and the sterile water extract between 1.25% and 10%. Its free radical scavenging rate IC50Values are shown in Table 7, IC50Lower values indicate better radical scavenging in this respect.
TABLE 7 DPPH radical scavenging ratio IC50Value of
Figure BDA0003154862180000131
The results show that the peony streptococcus thermophilus fermentation liquor and the peonyDpph free radical scavenging activity (IC) of lactobacillus plantarum fermentation broth50The values are respectively 1.41% and 1.37%), which are obviously higher than the water extract of peony and the water extract of sterilized peony (IC)50The values are 2.45% and 1.74%, respectively), and the removal effect of the two fermentation liquors is not obvious. The reason for this analysis may be: after the peony flower extract is subjected to high-pressure heating and liquid fermentation, plant cell walls of the peony flower extract are damaged and are easy to crack, and enzyme systems secreted by microorganisms can degrade cell walls, so that active ingredients such as flavone and polyphenol in the peony flower extract are easy to dissolve out, the content of the active ingredients is improved to different degrees, and the flavone and the polyphenol have oxidation resistance, so that the oxidation resistance of the peony flower extract is obviously improved after fermentation.
(3) Measurement of hydroxyl radical scavenging Rate
The determination results of the hydroxyl radical clearance rates of the peony water extract, the sterilized peony water extract, the peony streptococcus thermophilus fermentation broth and the peony lactobacillus plantarum fermentation broth are shown in fig. 8, the hydroxyl radical clearance rates of the extract and the fermentation broth show a tendency of decreasing with the decrease of the concentration, and the hydroxyl radical clearance rates of the peony streptococcus thermophilus fermentation broth and the peony lactobacillus plantarum fermentation broth are higher than those of the peony water extract and the sterilized water extract. Its hydroxyl radical scavenging rate IC50The values are shown in Table 8.
TABLE 8 hydroxyl radical scavenging ratio IC50Value of
Figure BDA0003154862180000141
The results show that the hydroxyl radical scavenging ability (IC) of the peony plant lactobacillus fermentation broth and the peony streptococcus thermophilus fermentation broth50The values are respectively 12.27 percent and 12.50 percent, which are obviously higher than the water extract of peony and the sterilized water extract (IC)50Values 21.31%, 18.07%, respectively). The hydroxyl radical scavenging capacity of the two fermentation liquors has no obvious difference.
(4) Determination of ABTS radical scavenging Rate
Peony water extract, sterile peony water extract, peony streptococcus thermophilus fermentation broth and peony plant lactobacillusThe measurement result of fermentation broth ABTS free radical clearance is shown in FIG. 9, and its ABTS free radical clearance IC50The values are shown in Table 9.
TABLE 9 ABTS radical clearance IC50Value of
Figure BDA0003154862180000142
The results show that the ABTS free radical clearance of all samples shows a trend of increasing and then decreasing with decreasing concentration. The clearance rates of the fermentation liquor of the peony streptococcus thermophilus and the fermentation liquor of the peony plant lactobacillus are higher than that of an aqueous extract and a sterilized aqueous extract when the clearance rates of the fermentation liquor of the peony streptococcus thermophilus and the fermentation liquor of the peony plant lactobacillus are between 0.16% and 0.63% and above 10%, wherein the clearance rates of the fermentation liquor of the peony plant lactobacillus and the fermentation liquor of the peony streptococcus thermophilus (IC)50The values are 0.295 percent and 0.296 percent), and the hydroxyl radical scavenging capacity is obviously higher than that of the peony water extract and the sterilized peony water extract (IC)50Values of 0.416%, 0.362%). Compared with fermentation liquor of different strains, the ABTS free radical scavenging capacity of the lactobacillus plantarum fermentation liquor is not greatly different from that of the streptococcus thermophilus fermentation liquor.
(5) Determination of tyrosinase inhibitory Effect
The determination results of tyrosinase inhibition effects of water extractive solution of flos moutan, sterile water extractive solution of flos moutan, Streptococcus thermophilus fermentation broth of flos moutan, and Lactobacillus plantarum fermentation broth of flos moutan are shown in FIG. 10, and the tyrosinase inhibition rate IC is shown in FIG. 1050The values are shown in Table 10.
TABLE 10 tyrosinase inhibition IC50Value of
Figure BDA0003154862180000151
The results show that the tyrosinase inhibition rates of all samples show a tendency to decrease with decreasing concentration, and the inhibition rates of both fermentation liquors are higher than those of the aqueous extract. Peony plant lactobacillus fermentation broth and peony streptococcus thermophilus fermentation broth (IC)50Values of 1.325% and 1.374%) are obviousHigher than peony water extract (IC)50Value of 2.435%) and sterile water extract (IC)50The value is 1.988%), and the tyrosinase inhibition effect of the peony plant lactobacillus fermentation liquor and the peony streptococcus thermophilus fermentation liquor is higher than that of the sterilized water extract.
It should be noted that lactobacillus plantarum (strain deposit No. CICC 20265) and streptococcus thermophilus (strain deposit No. CICC 20370) used in the present invention were purchased from china industrial microorganism culture collection.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Claims (10)

1. A microbial fermentation method for improving the content of active ingredients in peony is characterized by comprising the following steps:
s1, drying and crushing fresh peony into peony powder, mixing with distilled water, and performing ultrasonic treatment to obtain a mixed solution for later use;
s2, sterilizing and cooling the mixed solution obtained in the step S1, inoculating probiotics, performing fermentation culture, centrifuging, and taking supernate to obtain peony fermentation liquor;
the probiotics are lactobacillus plantarum or streptococcus thermophilus;
the fermentation conditions were: the inoculation amount is 1-10 vol%, the temperature is 36-37 ℃, the fermentation time is 12-48h, and the initial pH of the fermentation liquid is 5-7.
2. The microbial fermentation method for increasing the content of active ingredients in peony of claim 1, wherein in S2, the fermentation conditions are as follows: the inoculation amount is 1 vol%, the fermentation time is 36h, and the initial pH of the fermentation liquid is 6.5.
3. The microbial fermentation method for increasing the content of active ingredients in peony of claim 1, wherein in S1, the ratio of the peony powder to distilled water is 7.5 g: 140 and 160 mL.
4. The microbial fermentation method for increasing the content of active ingredients in peony of claim 1, wherein in S1, the drying condition is vacuum drying at 50 ℃.
5. The microbial fermentation method for increasing the content of active ingredients in peony according to claim 1, wherein in S1, the ultrasonic treatment condition is ultrasonic treatment at 60 ℃ for 30 min.
6. The microbial fermentation method for increasing the content of active ingredients in peony of claim 1, wherein in S2, centrifugation is performed at 10000r/min for 10 min.
7. A peony fermentation broth prepared by the microbial fermentation method for increasing the content of active ingredients in peony of claim 1.
8. Use of the peony fermentation broth of claim 7 in the preparation of a skin care product.
9. Use of the peony fermentation broth in the preparation of skin care products according to claim 8, wherein the peony fermentation broth is used for preparing antioxidant skin care products.
10. Use of the peony fermentation broth in the preparation of skin care products according to claim 8, wherein the peony fermentation broth is used for preparing skin care products with whitening effect.
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CN113925805A (en) * 2021-09-14 2022-01-14 上海植纳生物科技有限公司 Peony cell fermentation liquor and preparation method and application thereof
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CN114712282A (en) * 2022-04-07 2022-07-08 洛阳师范学院 Peony skin-care antibacterial composition and skin-care antibacterial liquid soap prepared from same
CN115251203A (en) * 2022-07-27 2022-11-01 中华全国供销合作总社济南果品研究所 Application and method of Xie Washi aspergillus in fermentation preparation of peony petal tea
CN115251203B (en) * 2022-07-27 2024-07-05 中华全国供销合作总社济南果品研究所 Application of Xie Washi aspergillus in preparing peony petal tea by fermentation and method
CN115067516A (en) * 2022-08-23 2022-09-20 山东凤凰生物科技股份有限公司 Application of lactobacillus plantarum BLCC2-0015, peony fermentation broth and application thereof
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CN115671024A (en) * 2022-12-07 2023-02-03 花安堂生物科技集团有限公司 Cherry flower glabridin composite fermentation liquid, product containing same, preparation and application thereof
CN116035975A (en) * 2023-02-10 2023-05-02 菏泽牡丹产业技术研究院 Application of peony flower water in skin care product
CN116035975B (en) * 2023-02-10 2024-01-26 菏泽牡丹产业技术研究院 Application of peony flower water in skin care product

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