CN113813203B - Cherry blossom enzymolysis fermentation product and preparation method and application thereof - Google Patents
Cherry blossom enzymolysis fermentation product and preparation method and application thereof Download PDFInfo
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Dermatology (AREA)
- Engineering & Computer Science (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Cosmetics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The application relates to the technical field of microbial fermentation, and discloses a sakura enzymolysis fermentation product, a preparation method and application thereof. The sakura enzymolysis ferment is prepared by pretreating sakura by enzyme liquid of aspergillus niger and aspergillus oryzae and fermenting by lactobacillus and saccharomycetes. According to the application, the enzymatic hydrolysis pretreatment of the sakura/jasmine is performed by using the enzymatic liquids of the aspergillus oryzae and the aspergillus niger, and then the sakura/jasmine is fermented by using specific lactic acid bacteria and microzyme, so that the obtained sakura/jasmine enzymatic fermentation product can remarkably improve the antioxidant activity, the anti-aging activity and the total flavone content, wherein the sakura enzymatic fermentation product is better, and a better foundation is provided for the application of the sakura/jasmine enzymatic fermentation product in the cosmetic field.
Description
Technical Field
The application relates to the technical field of microbial fermentation, in particular to a sakura enzymolysis fermentation product, a preparation method and application thereof.
Background
Sakura is a collective term for flowers of plants of the genus sakura of the family rosaceae. The main species of sakura in China are distributed in western and southwest regions of China. The application value of the sakura is more important in view of ornamental aspects and the medicinal value. The sakura has good effects of shrinking pores and balancing grease; in addition, the skin care product also contains rich natural vitamins A, B, E, and the cherry leaf flavone has the effects of maintaining beauty and keeping young, strengthening mucous membrane and promoting sugar metabolism, and has the effects of tendering the skin and brightening the skin, so that the cherry leaf flavone becomes one of important raw materials of the skin care product.
Jasmine (academic: jasminum sambac (L.) Ait) is a evergreen shrub of the genus Itopart. The Bay area of original production, the Han dynasty, was previously introduced into China with the communication of the middle and outer economies and cultures. The jasmine is fragrant in fragrance, elegant and pure, is deeply favored by people, and starts to be used as a medicinal plant from the open generation. In the "compendium of materia medica", the characters, cultivation and utilization of jasmine flower are described in more detail, and the jasmine flower is pointed out that "pungent heat is nontoxic", and can be used as "oil-steaming and liquid-taking, as face fat and head gloss, and long-lasting and dryness-moistening fragrance muscle". The jasmine flower extract has the main functions of resisting oxidation, whitening and removing freckles and promoting skin permeation, can eliminate free radicals, has strong penetrability and helps other nutrient components to be absorbed by skin; has certain tyrosinase inhibiting effect, and can assist in whitening. Because of its faint scent, it is commonly used in cosmetics for preparing flavors and fragrances.
At present, the extraction of the active ingredients in the flowers mainly adopts an alcohol reflux extraction technology, an ultrasonic extraction technology, a physical and chemical extraction means such as a desorption-thermal extraction two-step method and the like, and the extraction report of the active ingredients by utilizing a microbial fermentation technology is less, and the synergistic effect of different flower extracts is not clear.
Disclosure of Invention
Therefore, the application aims to provide the sakura enzymolysis ferment and the preparation method thereof, so that the prepared sakura enzymolysis ferment can obviously improve the antioxidation capability;
the application also aims to provide the sakura enzymolysis fermented product and the preparation method thereof, so that the prepared sakura enzymolysis fermented product can obviously improve the anti-aging activity;
the application also aims to provide the sakura enzymolysis ferment and the preparation method thereof, so that the prepared sakura enzymolysis ferment can obviously improve the total flavone content;
another object of the present application is to provide an application of the above-prepared enzymatic hydrolysate of sakura in preparing cosmetics;
it is another object of the present application to provide a plant combination ferment comprising the above-prepared sakura enzymolysis ferment and an application thereof in preparing cosmetics.
In order to solve the technical problems or at least partially solve the technical problems, the application provides a sakura enzymolysis ferment which is prepared by fermenting sakura through lactobacillus and saccharomycetes after being pretreated by enzyme liquid of aspergillus niger and aspergillus oryzae.
In a specific embodiment of the application, the yeast is Mark Lu Wei yeast, which is available from China industry microbiological culture Collection center, and is numbered CICC 1218. The lactobacillus is Lactobacillus plantarum, specifically Lactobacillus plantarum subspecies, and is numbered CICC 20242. Aspergillus niger and Aspergillus oryzae in the enzymatic pretreatment stage of the application are also available from the China center for type culture Collection of microorganisms, and are numbered CICC 2039 and CICC 2005.
Preferably, the fermentation enzyme liquid of the aspergillus niger is obtained by solid fermentation of the aspergillus niger in a culture medium containing a carbon source, a nitrogen source, inorganic salt and jasmine. Wherein the carbon source and nitrogen source are orange peel and bran, and the inorganic salt is selected from MS, HB, LS, white inorganic salt such as (NH) 4 ) 2 SO 4 、K 2 HPO 4 、KCl、MgSO 4 、Na 2 SO 4 、FeSO 4 Etc. or their hydrates.
Preferably, the Aspergillus oryzae fermenting enzyme liquid is obtained by solid fermentation of Aspergillus oryzae in a mixture of Oryza Glutinosa and water.
Compared with the product obtained by adopting jasmine enzymolysis and fermentation, the cherry enzymolysis and fermentation product prepared by the application has obviously improved DPPH clearance and anti-aging activity, and simultaneously has obviously improved total flavone content. Therefore, the application provides application of the sakura enzymolysis fermentation product in preparing cosmetics, wherein the cosmetics are cosmetics which are required for additionally improving or enhancing whitening, freckle removing, anti-aging, antioxidation and other effects.
Meanwhile, the application also provides a preparation method of the sakura enzymolysis ferment, which comprises the following steps:
step 1, inoculating aspergillus niger and aspergillus oryzae into an enzyme-producing culture medium for solid fermentation to obtain an enzyme solution;
step 2, performing enzymolysis pretreatment on the sakura with an enzyme solution of aspergillus niger and aspergillus oryzae;
and 3, fermenting the enzymolysis pretreatment liquid by using lactobacillus and saccharomycetes to obtain the sakura enzymolysis fermentation product.
Preferably, the step 1 is:
inoculating Aspergillus niger into a culture medium containing a carbon source, a nitrogen source, inorganic salt and jasmine flower for solid fermentation, and adding citric acid buffer solution (preferably 1:20 mass-volume ratio) ice into the fermentation liquidBath extraction, centrifuging and taking supernatant to obtain Aspergillus niger enzyme solution; further preferably, the medium comprises citrus peel, bran, jasmine, (NH) 4 )2SO 4 、K 2 HPO 4 、KCl、MgSO 4 、Na 2 SO 4 、FeSO 4 The method comprises the steps of carrying out a first treatment on the surface of the In a specific embodiment, the medium comprises the following components per 1L:
2.5g of citrus peel, 1g of bran and 1g of jasmine, (NH) 4 ) 2 SO 4 1g，K 2 HPO 4 0.04g，KCl 0.03g，MgSO 4 ·7H 2 O 0.03g，Na 2 SO 4 0.02g，FeSO 4 ·7H 2 0.002g of O, adjusting the pH value to 4.5, adding a proper amount of purified water to a constant volume of 1L, and sterilizing for 20min at 121 ℃.
Inoculating Aspergillus oryzae into a mixture of Oryza Glutinosa and water for solid fermentation, adding citric acid buffer solution (preferably 1:20 mass/volume ratio) into the fermented solution, extracting in ice bath, centrifuging, and collecting supernatant to obtain Aspergillus oryzae enzyme solution. Further preferably, the glutinous rice: water = 30:36 (mass ratio); natural pH, sterilizing at 100deg.C for 60min; citric acid buffer ph4.8;
preferably, the enzymolysis pretreatment in the step 2 is carried out at a temperature within a proper range of the enzyme solution, for example, 50 ℃ +/-5 ℃;
preferably, the step 3 is as follows:
inoculating lactobacillus to the enzymolysis pretreatment liquid for fermentation in the first stage, inoculating saccharomycetes for fermentation in the second stage after fermentation, and centrifuging the fermentation liquid to obtain supernatant serving as the jasmine enzymolysis fermentation product. Further preferably, the two stages of fermentation time is 7d, the lactobacillus inoculation proportion is 5%, the yeast inoculation proportion is 1%, the fermentation environment is selected to be a proper environment of the strain, and a temperature range of 37+/-5 ℃ is provided in the implementation of the application.
In addition, in view of the excellent performance of the sakura enzymolysis fermentation product, the application also provides a plant composition fermentation product containing the sakura enzymolysis fermentation product, which is used for being combined with other components such as plant extracts, plant fermentation products and the like which can be used for cosmetics on the basis of the sakura enzymolysis fermentation product. Preferably, the plant combination fermentation product provided by the application comprises a sakura enzymolysis fermentation product and a jasmine enzymolysis fermentation product, wherein the preferable mass ratio of the sakura enzymolysis fermentation product to the jasmine enzymolysis fermentation product is 1-2:1-2; the jasmine enzymolysis fermentation product is prepared by fermenting jasmine after being pretreated by enzyme liquid of aspergillus niger and aspergillus oryzae by lactic acid bacteria and saccharomycetes. As the best scheme of the plant combination ferment, the preparation of the jasmine enzymolysis ferment can refer to the preparation process of the cherry enzymolysis ferment.
In the optimal scheme, indexes such as antioxidant capacity and the like of the jasmine enzymolysis fermentation product are researched and compared, compared with other fermentation products of different strains and different preparation processes and jasmine alcohol extracts, the jasmine enzymolysis fermentation product obtained by the application has obviously improved DPPH clearance, ABTS clearance, hydroxyl radical clearance and superoxide anion clearance, and also has obviously improved crude polysaccharide and total flavone contents. Based on the above, the application also provides application of the plant combination fermentation product in preparing cosmetics, wherein the cosmetics are cosmetics which are required for additionally improving or enhancing whitening, freckle removing, anti-aging, antioxidant and other effects.
According to the technical scheme, the fermentation enzyme liquid of aspergillus oryzae and aspergillus niger is used for carrying out enzymolysis pretreatment on sakura/jasmine, and then specific lactic acid bacteria and saccharomycetes are used for fermenting the sakura/jasmine, so that the obtained sakura/jasmine enzymolysis fermented product can obviously improve the antioxidant activity, the anti-aging activity and the total flavone content, wherein the sakura enzymolysis fermented product is better, and a better foundation is provided for the application of the sakura/jasmine enzymolysis fermented product in the cosmetic field.
Detailed Description
The application discloses a cherry blossom enzymolysis ferment, a preparation method and application thereof, and a person skilled in the art can properly improve the technological parameters by referring to the content of the application. It is expressly noted that all such similar substitutions and modifications will be apparent to those skilled in the art, and are deemed to be included in the present application. While the process, application, and product of the present application have been described in terms of preferred embodiments, it will be apparent to those skilled in the relevant art that the process, application, and product of the present application can be modified or adapted to implement and use the teachings of the present application without departing from the spirit, scope, and range of equivalents of the application. It will be apparent that the described embodiments are some, but not all, embodiments of the application. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
It should be noted that in this document, relational terms such as "first" and "second," "step 1" and "step 2," and "(1)" and "(2)" and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.
In the comparison experiment of the application, except for due differences of all groups, other experimental environments, technological parameters, raw material source batches and the like which are not specifically described are kept consistent, so that the comparability of experimental results is ensured.
1. Preparation of enzyme solution for pretreatment of cherry/jasmine enzymolysis in the application (Aspergillus niger enzyme solution: aspergillus oryzae enzyme solution=2:1)
(1) Aspergillus niger enzyme production method:
aspergillus niger CICC 2039, china center for type culture Collection of Industrial microorganisms;
enzyme-producing medium: 2.5g of citrus peel, 1g of bran and 1g of jasmine, (NH) 4 ) 2 SO 4 1g，K 2 HPO 4 0.04g，KCl 0.03g，MgSO 4 ·7H 2 O 0.03g，Na 2 SO4 0.02g，FeSO 4 ·7H 2 0.002g of O, adjusting the pH value to 4.5, adding a proper amount of purified water to a constant volume of 1L, and sterilizing for 20min at 121 ℃.
Carrying out solid fermentation, adding citric acid buffer solution (1:20 mass/volume ratio) into the fermentation finished solution, carrying out ice bath extraction, centrifuging to obtain supernatant, obtaining fermentation, centrifuging the fermentation solution, and obtaining supernatant to obtain Aspergillus niger enzyme solution.
(2) The method for producing the enzyme by the aspergillus oryzae comprises the following steps:
aspergillus oryzae: CICC 2005, china center for type culture Collection of Industrial microorganisms;
enzyme-producing medium: glutinous rice: water=30:36 (mass ratio); natural pH, sterilizing at 100deg.C for 60min.
Mixing the activated Aspergillus oryzae seed liquid according to 10 6 Inoculating the spores/g of glutinous rice into a glutinous rice solid enzyme-producing culture medium, and standing and culturing at 30 ℃ for 72 hours. Preparing a citric acid buffer solution with pH of 4.8, adding the citric acid buffer solution according to a mass-volume ratio of 1:20, carrying out ice bath overnight, centrifuging, and taking the supernatant as Aspergillus oryzae crude liquid.
(3) The monascus enzyme production method comprises the following steps:
monascus cic cc40941 collection of industrial microbial strains in china; the method is the same as that of Aspergillus niger.
2. Fermentation strain and activation thereof
Lactic acid bacteria: lactobacillus plantarum (lactobacillus plantarum subspecies CICC 20242);
yeast: saccharomyces cerevisiae (BNCC 188427, north Nanoorganism, strain for comparison), saccharomyces marxianus Lu Wei (Kluyveromyces marxianus CICC 1218);
yeast: inoculating strain into YPD culture medium, culturing at 30deg.C for 36-48 hr, and continuously passaging for 3 times to fully activate strain until viable count is 1.0X10 7 CFU/mL or more;
lactic acid bacteria: inoculating strain into MRS culture medium, culturing at 35deg.C for 24 hr, and continuously passaging for 3 times to fully activate strain until viable count is 1.0X10 8 CFU/mL or more.
3. Enzymolysis fermentation mode
Aspergillus niger and Aspergillus oryzae fermentation broth: jasmine flower/sakura dry powder=20:1 ratio, after enzymolysis for 3 hours at 50 ℃, adding 4 times of volume of water, subpackaging in 250mL blue cap reagent bottles (200 mL/bottle), sterilizing (115 ℃ for 30 min) and preparing for fermentation. Firstly, inoculating activated lactobacillus strains according to the proportion of 5% of inoculum size, uniformly mixing, placing in an incubator at 37 ℃, fermenting for 7 days, inoculating saccharomycetes according to the proportion of 1%, fermenting for 7 days, and centrifuging for 15min at 9000r/min to obtain jasmine enzymolysis fermentation product or sakura enzymolysis fermentation product for index detection.
4. Index detection method
(1) Detection of crude polysaccharide content: the total content of crude polysaccharide in the fermentation broth and extract was determined with reference to SN/T4260-2015.
(2) And (3) detecting the total flavone content: the total flavone content in the fermentation broth and the extract was determined by the method described in GB/T12143-2008 appendix G.
(3) Measurement of DPPH radical scavenging ability:
the experimental group was added with 1ml of the sample solution+1 ml of DPPH at 0.2 mmol/L. Control group was added with 1ml absolute ethanol+1 ml stock solution. The blank was added with 1ml of absolute ethanol+1 ml of DPPH at 0.2mmol/L, and the mixture was allowed to stand at room temperature (25 ℃) for 30 minutes. At 517nm, 300ul of the sample was aspirated, and the absorbance of the test group was measured and recorded as Ai. The absorbance of the control group was measured and designated Aj. Blank set is denoted as A0. Each sample was measured 3 times in parallel.
DPPH clearance (%) = [1- (Ai-Aj)/A0 ] ×100%
(4) Superoxide anion radical scavenging ability assay:
5.7mL of Tris-HCl buffer solution (50 mmol/L, pH 8.2) is added into a 10mL test tube, 0.2mL of sample is added for mixing, the mixture is placed in a 25 ℃ incubator, after 10min, the mixture is taken out, 0.1mL (10 mmol/L) of pyrogallol solution (preheated) is added, after rapid mixing, the increase value (Aj) of absorbance within 1min at 320nm wavelength is measured by a multifunctional enzyme-labeling instrument, the increase value of absorbance per 1min is calculated within a linear range, the reagent is taken out, equal volume of water is used for replacing the sample, and the increase value (Ai) of absorbance within 1min at 320nm wavelength is measured.
Superoxide anion radical ion radical clearance (%) = (Ai-Aj)/ai×100%
(5) Determination of the scavenging ability of hydroxyl radicals (. OH):
hydroxyl radical (. OH) scavenging test method referring to the Fenton reaction method, ferrous sulfate solution (6 mmol/L), hydrogen peroxide solution (6 mmol/L) and 2mL of each sample are sequentially added into a test tube, and then the test tube is left standing for 10min, 2mL of salicylic acid solution (6 mmol/L) is added, and after standing for 30min, absorbance A0 is measured at 510nm, distilled water is used for replacing the sample solution to perform the same treatment, and the absorbance Ax is measured.
Hydroxyl radical (·oh) clearance (%) = (1-A0/Ax) ×100%
(6) Determination of ABTS clearance:
0.2mL of 7.4mmoL/L ABTS was mixed with 0.2mL of 2.6mmoL/L K S2O8, reacted at room temperature in the dark for 12 hours, and after the reaction was completed, diluted 40-50 times with 95% ethanol (pH=7.4 phosphate buffer, 95% ethanol or methanol) so that the absorbance of the mixed solution at 734nm was 0.68-0.72 (working solution).
Preparing and sucking 0.2mL of Vc standard solution with the concentration of 0 mug/mL, 4 mug/mL, 8 mug/mL, 12 mug/mL, 16 mug/mL and 20 mug/mL into a 2mL centrifuge tube, adding 0.8mL of diluted working solution, uniformly mixing, standing for reaction for 6-8min, and rapidly measuring the absorbance at 734nm wavelength. And drawing a standard curve by taking the concentration of the standard solution as an abscissa and the absorbance value as an ordinate. And (3) respectively sucking 0.2mL of diluted enzyme samples after diluting the different treatment enzyme samples, adding 0.8mL of diluted working solution, uniformly mixing, standing for reaction for 6-8min, and rapidly measuring the absorbance at the wavelength of 734 nm. Each sample was measured 3 times in parallel.
(7) And (3) anti-aging capability detection:
in order to determine whether the flower fermentation broth HAs anti-aging capability, in vitro level anti-aging evaluation is carried out on the flower fermentation broth, and the levels of matrix metalloproteinase-1 (MMP-1) secreted by HFF-1 fibroblasts, human L-hydroxyproline (L-HyP), human type I collagen (Col-I) and human Hyaluronic Acid (HA) are detected.
Collecting HFF-1 cells grown to more than 80% in the flask at 1×10 per well 4 The cell suspension with the concentration of one/mL is paved on a 96-well plate, then is placed in a constant temperature incubator at 37 ℃ for culturing for 24 hours, and then flower enzymolysis liquid or fermentation liquid which is diluted 10 times is added. All reagents and samples were equilibrated slowly to room temperature (18-25 ℃) before use, and the reagents could not be dissolved directly at 37 ℃. Culturing cell supernatantCentrifuging the nutrient solution at the temperature of 2-8 ℃ for 15min at the speed of 1000r/min, and taking the supernatant as a sample to be detected. The concentration of MMP-1, L-HyP, col-I, HA in the supernatants of HFF-1 cells of each experimental group was calculated by measuring OD450 using a 96-well plate microplate reader according to the methods described in the specifications of human MMP-1, L-HyP, col-I and HA.
Example 1: the experimental groups and the results of the detection of the oxidation resistance and the anti-aging activity of the flower enzymolysis fermented product and the combined fermented product are shown in the following table 1;
TABLE 1
Note that: the jasmine flower/cherry flower zymolyte is prepared by only adopting Aspergillus niger and Aspergillus oryzae zymolyte for enzymolysis, centrifuging, taking supernatant for detection, and not carrying out subsequent fermentation links;
the DPPH clearance rate and the total flavone content result show that the jasmine flower/sakura enzymolysis fermented product and the combined fermented product of the jasmine flower/sakura enzymolysis fermented product are obviously higher than those of the jasmine flower/sakura enzymolysis fermented product group;
meanwhile, after the jasmine flower/sakura enzymolysis fermented product and the combined fermented product of the jasmine flower/sakura enzymolysis fermented product and the sakura enzymolysis fermented product are diluted by 10 times, HFF-1 cells are cultured, and the MMP-1 content is obviously lower than that of a blank control group and a jasmine flower/sakura enzymolysis fermented product group. Thus, it was demonstrated that each group achieved an anti-aging effect by inhibiting the secretion of MMP-1 by HFF-1 cells. The L-HyP concentration, HA concentration and Col-I concentration of each group in HFF-1 cells are obviously higher than those of the blank group and the jasmine flower/sakura zymolyte group. In the HFF-1 extracellular matrix assay, each group had anti-aging ability to promote secretion of L-HyP, HA, col-I by cells.
Among the above groups, the group effect related to the enzymatic fermentation of cherry blossom is better.
Example 2: antioxidant capacity of jasmine enzymolysis ferment of different processes and comparison of total flavone and crude polysaccharide content
The following enzymolysis fermentation modes of different treatments are compared, and the grouping scheme is as follows (5 g of jasmine flower dry powder in each group):
(1) Jasmine enzymolysis pretreatment
Enzyme liquid obtained by fermenting Aspergillus niger and Aspergillus oryzae: jasmine flower dry powder=20:1 ratio, extracting at 50 ℃ for 3 hours, centrifuging, and taking supernatant for index detection.
(2) Alcohol reflux extraction of jasmine
Weighing 5g of jasmine flower dry powder, precisely weighing, placing into a round bottom flask, adding 50-90% ethanol 50mL, reflux-extracting at 80deg.C, taking out liquid every 1h, filtering, continuously adding 50-90% ethanol 50mL into the residue, reflux-extracting, and repeating the above steps for 3 times.
(3) Jasmine flower direct fermentation (lactobacillus + Saccharomyces cerevisiae)
Taking jasmine flower dry powder, adding water according to the mass-volume ratio of 1:100, subpackaging in 250mL blue-cap reagent bottles (200 mL/bottle), sterilizing (115 ℃ C., 30 min), and preparing for fermentation. Firstly, inoculating activated lactobacillus strains according to the proportion of 5% of the inoculum size, uniformly mixing, placing in an incubator at 37 ℃, fermenting for 7 days, inoculating Saccharomyces cerevisiae according to the proportion of 1%, fermenting for 7 days, and centrifuging for 15min at 9000r/min to obtain jasmine flower fermented product for index detection.
(4) Jasmine flower direct fermentation (lactobacillus + Ma Kelu vitamin yeast)
In the same way as in the process of the group (3), the yeast is selected from the yeast of Mark Lu Wei;
(5) Jasmine enzymolysis pretreatment (Aspergillus oryzae+Aspergillus niger) +fermentation
Enzyme liquid obtained by fermenting Aspergillus niger and Aspergillus oryzae: jasmine flower dry powder=20:1 ratio, after enzymolysis for 3 hours at 50 ℃, adding water with 4 times of volume, subpackaging in 250mL blue-cap reagent bottles (200 mL/bottle), sterilizing (115 ℃ for 30 min) and preparing for fermentation. Firstly, inoculating activated lactobacillus strains according to the proportion of 5% of inoculum size, uniformly mixing, placing in an incubator at 37 ℃, fermenting for 7 days, inoculating Kluyveromyces marxianus according to the proportion of 1%, fermenting for 7 days, and centrifuging for 15min at 9000r/min to obtain jasmine flower fermented product for index detection.
(6) Fermenting all strains of jasmine together
Taking jasmine flower dry powder, adding water according to the mass-volume ratio of 1:100, subpackaging in 250mL blue-cap reagent bottles (200 mL/bottle), sterilizing (115 ℃ C., 30 min), and preparing for fermentation. Firstly inoculating activated Aspergillus oryzae and Aspergillus niger strains according to the proportion of 5% of inoculum size, culturing in an incubator at 28 ℃ for 7 days, inoculating lactobacillus strains according to the proportion of 5% of inoculum size, mixing uniformly, fermenting in an incubator at 37 ℃ for 7 days, inoculating Saccharomyces cerevisiae according to the proportion of 1%, fermenting at 30 ℃ for 7 days, and centrifuging at 9000r/min for 15min to obtain jasmine fermentation product for index detection.
(7) Jasmine enzymolysis pretreatment (aspergillus oryzae) +fermentation
Referring to the process of group (5), the difference is that Aspergillus oryzae is used entirely;
(8) Jasmine enzymolysis pretreatment (Aspergillus niger) +fermentation
Referring to the process of group (5), the difference is that all Aspergillus niger is used;
(9) Jasmine enzymolysis pretreatment (Aspergillus oryzae+monascus) +fermentation
Referring to the process of group (5), the difference is that Aspergillus niger is replaced with Monascus purpureus;
the detection results of the indexes of each group are shown in Table 2;
TABLE 2
As can be seen from the results of Table 1, the pretreatment with Aspergillus oryzae and Aspergillus niger enzymes and the subsequent fermentation gave higher antioxidant capacity, crude polysaccharide content and total flavone content, whereas the control group, which was set by some means or similar microorganism strains in the prior art, had a certain antioxidant capacity, but was clearly inferior to the enzymatic jasmine fermented product of the present application (group 5 process), and the alcoholic extract of jasmine was not as whole as the microbial fermentation process.
In addition, the application detects the DPPH clearance rate of the process of the group (5) by 50 times of stock solution and dilution, and the DPPH clearance rate is about 99 percent and 95 percent respectively.
The foregoing is only a specific embodiment of the application to enable those skilled in the art to understand or practice the application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the application. Thus, the present application is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (5)
1. The preparation method of the sakura enzymolysis ferment is characterized by comprising the following steps:
step 1, inoculating Aspergillus niger of CICC 2039 to a mixture comprising pericarp, bran, flos Jasmini sambac, (NH) 4 ) 2 SO 4 、K 2 HPO 4 、KCl 、MgSO 4 、Na 2 SO 4 、FeSO 4 Performing solid fermentation in the culture medium of (1), adding citric acid buffer solution into the fermentation broth, performing ice bath extraction, centrifuging, and taking the supernatant to obtain Aspergillus niger enzyme solution; inoculating Aspergillus oryzae of CICC 2005 into a mixture of Oryza Glutinosa and water for solid fermentation, adding citric acid buffer solution after fermentation, extracting in ice bath, centrifuging, and collecting supernatant to obtain Aspergillus oryzae enzyme solution;
step 2, performing enzymolysis pretreatment on the sakura with enzyme solutions of aspergillus niger and aspergillus oryzae at 50+/-5 ℃; wherein, aspergillus niger enzyme liquid: aspergillus oryzae enzyme liquor = 2:1, aspergillus niger and Aspergillus oryzae fermentation enzyme liquor: sakura = 20:1;
step 3, inoculating lactobacillus plantarum of CICC 20242 into the enzymolysis pretreatment liquid for fermentation in the first stage, inoculating the microzyme of Mark Lu Wei of CICC 1218 for fermentation in the second stage after the fermentation is finished, and centrifuging the fermentation liquid to obtain supernatant which is the sakura enzymolysis fermentation product; the fermentation time of the two stages is 7d, and the fermentation temperature is 37+/-5 ℃.
2. The cherry blossom enzymolysis fermented product is characterized by being prepared by the preparation method according to claim 1.
3. Use of the enzymatic ferment of sakura in accordance with claim 2 for the preparation of anti-aging and anti-oxidative cosmetics.
4. A plant composite ferment, characterized by comprising the sakura enzymolysis ferment of claim 2, and a jasmine enzymolysis ferment, a jasmine enzymolysis ferment: the mass ratio of the cherry blossom enzymolysis ferment is 1:2;
the jasmine enzymolysis fermentation product is prepared by the following preparation method:
step 1, inoculating Aspergillus niger of CICC 2039 to a mixture comprising pericarp, bran, flos Jasmini sambac, (NH) 4 ) 2 SO 4 、K 2 HPO 4 、KCl 、MgSO 4 、Na 2 SO 4 、FeSO 4 Performing solid fermentation in the culture medium of (1), adding citric acid buffer solution into the fermentation broth, performing ice bath extraction, centrifuging, and taking the supernatant to obtain Aspergillus niger enzyme solution; inoculating Aspergillus oryzae of CICC 2005 into a mixture of Oryza Glutinosa and water for solid fermentation, adding citric acid buffer solution after fermentation, extracting in ice bath, centrifuging, and collecting supernatant to obtain Aspergillus oryzae enzyme solution;
step 2, performing enzymolysis pretreatment on the sakura with enzyme solutions of aspergillus niger and aspergillus oryzae at 50+/-5 ℃; wherein, aspergillus niger enzyme liquid: aspergillus oryzae enzyme liquor = 2:1, aspergillus niger and Aspergillus oryzae fermentation enzyme liquor: jasmine = 20:1;
step 3, inoculating lactobacillus plantarum of CICC 20242 into the enzymolysis pretreatment liquid for fermentation in the first stage, inoculating the mark Lu Wei saccharomycete of CICC 1218 for fermentation in the second stage after the fermentation is finished, and centrifuging the fermentation liquid to obtain supernatant which is the jasmine enzymolysis fermentation product; the fermentation time of the two stages is 7d, and the fermentation temperature is 37+/-5 ℃.
5. Use of the plant combination ferment according to claim 4 for the preparation of anti-aging and anti-oxidation cosmetics.
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