Rose fermentation product and preparation method thereof, and extraction method and application of rose polysaccharide

Abstract

The invention relates to a rose fermentation product and a preparation method thereof, and an extraction method and application of rose polysaccharide, and belongs to the technical field of microbial fermentation. According to the method, a microbial fermentation mode is adopted, rose, rose leaves and rose roots are selected as main raw materials, red peony roots, trehalose and oat powder are used as auxiliary materials for extraction and fermentation to prepare the rose fermentation product, reaction conditions are mild, and no chemical reagent needs to be added. The invention also provides application of the rose fermentation product in preparation of whitening and anti-inflammatory products. According to the invention, each part in the rose is utilized, the effective components in the raw materials can be fully extracted, and the functional components and activity of plants can be relatively completely retained. Release of active ingredients of roses and synthesis of new substances are facilitated, and the content of small-molecule active substances in the product is higher.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a rose fermentation product and a preparation method thereof, and a rose polysaccharide extraction method and application thereof. Background Art

[0002] Rose is a plant of the genus Rosa in the family Rosaceae. Its petals are obovate, double or semi-double, and its stems are thorny and relatively soft. It is widely distributed around the world, with rich species and strong fragrance, and is more distributed in the northern hemisphere. Roses have been developed and utilized by people for a long time. Its flowers contain aromatic alcohols, aldehydes, fatty acids, flavonoids, phenols, and essential oils and fats, as well as sesquiterpenes, esters, polysaccharides, alkaloids and other substances, with a total of more than 300 chemical components. Rose essential oil, also known as rose water, is extracted from rose petals by distillation. It can effectively improve skin texture, promote blood circulation and metabolism, and it also has resistance to inflammation, tumors, oxidation, and toxin accumulation, and has a wide range of pharmacological activities.

[0003] Fermentation is a process in which microorganisms carry out physiological activities under certain conditions, decompose and transform organic matter with the help of enzymes to obtain various nutrients such as C, N, vitamins, etc. to grow bacteria, and produce various secondary metabolites, such as polysaccharides and amino acids. Compared with other processing methods, fermentation can produce new active substances, reduce toxicity, change efficacy, and at the same time, due to the hydrolysis of microorganisms, the fermentation products are easier to absorb.

[0004] In recent years, there have been many studies on rose fermentation, but most of them are aimed at the fermentation of rose flowers. There are fewer studies on rose leaves and roots, especially on the polysaccharides, polyphenols and monoterpenes obtained after fermentation of rose leaves and roots. Summary of the invention

[0005] In view of the problem that rose leaves and rose roots cannot be fully utilized in the prior art, the present invention provides a rose fermentation product and a preparation method, and a rose polysaccharide extraction method and application to solve the above problems. The present invention adopts a microbial fermentation method to prepare a fermentation liquid using rose leaves and rose roots, and the reaction conditions are mild, no chemical reagents need to be added, the product is highly safe, the fermentation liquid contains a large amount of fermentation enzymes, the effective components in the plant raw materials can be fully extracted, and the effective components and their activities of the plants can be relatively completely retained.

[0006] In a first aspect, the present invention provides a method for preparing a rose fermentation product, comprising the following steps: (1) Pretreatment: Weigh rose flowers, rose leaves, rose roots, red peony root, trehalose and oatmeal powder in proportion, crush, sieve, add to pure water, add pectinase and papain, and treat at 40-50°C for 30 min under stirring; sterilize at 105°C for 15-20 min to obtain rose culture medium; (2) Preliminary fermentation: Inoculate the fermented Lactobacillus rhamnosus seed liquid into rose culture medium and culture it in an anaerobic incubator at 37°C for 12-16 hours. Add 10% sodium hydroxide solution to adjust the pH to 6.0 to obtain the preliminary fermentation liquid; (3) Secondary fermentation: Inoculate the composite bacterial liquid of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus plantarum and Kluyveromyces marxianus into the primary fermentation liquid, incubate in an anaerobic incubator at 37°C for 24 hours, then adjust the temperature to 28-31°C and ferment for another 48-96 hours; (4) Decolorization: add 0.5% to 1.0% activated carbon based on the weight of the fermentation liquid, heat to 80°C, stir for 10 min, centrifuge at 10,000 rpm to obtain a centrifuge, filter with an 8-10 μm filter membrane to obtain a light yellow clear filtrate; (5) Concentration treatment: The clarified filtrate is concentrated to 50% by vacuum rotary evaporation equipment to obtain the rose fermentation product.

[0007] Furthermore, step (1) comprises the following raw materials in parts by weight: 10 parts of rose flowers, 3 parts of rose leaves, 1 part of rose roots, 1 part of red peony root, 1 part of trehalose, 1 part of oatmeal powder, and 200 parts of water.

[0008] Furthermore, in step (1), the amount of pectinase added is 0.5% to 0.6% based on the total weight of the raw materials; the enzyme activity is ≥5×10 4 U / g; the amount of papain added is 0.4%~0.6% based on the total weight of the raw materials, and the enzyme activity is ≥5×10 4 U / g.

[0009] Furthermore, in step (2), the bacterial concentration of the Lactobacillus rhamnosus fermentation seed liquid is ≥10 6 CFU / ml.

[0010] Furthermore, in step (2), the amount of Lactobacillus rhamnosus fermented seed liquid added is 3% to 5% by weight of the rose culture medium.

[0011] Furthermore, in the composite bacterial solution of step (3), the mass ratio of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus plantarum and Kluyveromyces marxianus is: 6 ~10 8 CFU / ml.

[0012] In a second aspect, the present invention provides a rose fermentation product prepared by the above preparation method, wherein the rose polysaccharide content in the rose fermentation product is ≥10.4 g / L, and the polyphenol content is ≥5.8 g / L.

[0013] In a third aspect, the present invention provides a method for extracting rose polysaccharide, the steps of which are as follows: add 1 part of composite protease to 100 parts of rose fermentation products, adjust the pH to 7-8, and treat at 37°C for 2-4 hours. After digestion, treat at 80°C for 2 minutes. Add to 3-4 parts of 95% ethanol, let stand for 12-24 hours, centrifuge at 12000-16000 r / min for 5-10 minutes, obtain a precipitate, and dry at 45-60°C to obtain rose polysaccharide.

[0014] In a fourth aspect, the present invention provides an application of a rose fermentation product in the preparation of whitening and anti-inflammatory products.

[0015] The beneficial effects of the present invention are: (1) The present invention selects rose flowers, rose leaves and rose roots as main raw materials, which is significantly different from the previous single extraction of rose flowers, rose leaves or rose roots, and uses red peony root, trehalose and oatmeal powder as auxiliary materials for extraction and fermentation, making comprehensive use of different parts of roses, which is more conducive to the release of rose active ingredients and the synthesis of new substances, and the product has higher small molecule active substances.

[0016] (2) The present invention adopts a step-by-step extraction method, and preferentially uses enzymatic hydrolysis technology, which can promote the breaking of plant cell walls and facilitate the release of intracellular substances. The use of rhamnosus lactobacillus fermentation can promote the production of small molecule proteins and enzyme substances, such as acid proteases and amylases, which can decompose plant cell tissues and produce rich metabolites. The fermentation of secondary composite strains and low-temperature fermentation are conducive to the growth of bacteria and the slow accumulation of metabolites. It produces richer polysaccharides and polyphenol components, making the product have higher anti-inflammatory and whitening properties, and can effectively promote cell proliferation and metabolism, promote cell regeneration, improve skin elasticity, and have good free radical scavenging effects, and have good anti-aging effects.

[0017] (3) The present invention adds a small amount of oatmeal powder to the culture medium, and undergoes two-step fermentation and low-temperature long-term fermentation, which is more conducive to the synthesis of polysaccharides. The polysaccharide content in the rose fermentation product can reach 10.5g / L, the polyphenol content can reach 5.8g / L, and the fermentation product has excellent moisturizing and lubricating properties, good moisturizing effects, and more advantageous antioxidant and anti-inflammatory effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0019] Figure 1 This is the fermentation product prepared in Example 1 of the present invention.

[0020] Figure 2 This is the rose polysaccharide prepared in Example 4 of the present invention. DETAILED DESCRIPTION

[0021] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0022] The strains and compound protease models and sources used in the embodiments of the present invention are as follows: (1) Lactobacillus rhamnosus: Model 01, Manufacturer: Shandong Jiacheng Biotechnology Co., Ltd., Purchase website: https: / / detail.1688.com / offer / 689387068656.html?_t=1733730748098&spm=a2615.7691456.co_0_0_wangpu_score_0_0_0_0_1_0_0000_0.0; (2) Bifidobacterium longum: model PG-BLC107, manufacturer: Shandong Jiacheng Biotechnology Co., Ltd., purchase website: https: / / detail.1688.com / offer / 689999622130.html?_t=1733730925363&spm=a2615.7691456.co_0_0_wangpu_score_0_0_0_0_1_0_0000_0.0; (3) Lactobacillus acidophilus: model PG-LAS107, manufacturer: Shandong Jiacheng Biotechnology Co., Ltd., purchase website: https: / / detail.1688.com / offer / 782779962392.html?_t=1733731507892&spm=a2615.7691456.co_0_0_wangpu_score_0_0_0_0_1_0_0000_0.0; (4) Lactobacillus plantarum: model PG-LPZ131, manufacturer: Shandong Jiacheng Biotechnology Co., Ltd., purchase website: https: / / detail.1688.com / offer / 724612644381.html?_t=1733731557399&spm=a2615.7691456.co_0_0_wangpu_score_0_0_0_0_1_0_0000_0.0; (5) Kluyveromyces marxianus: Manufacturer: Shandong Pingju Biotechnology Co., Ltd., purchase website: https: / / china.guidechem.com / trade / pdetail28156064.html; (6) Compound protease: Manufacturer: Jiangsu Ruiduo Bioengineering Co., Ltd., purchase website: https: / / b2b.baidu.com / land?url=https%3A%2F%2Fb2bwork.baidu.com%2Fland%3Flid%3D1760693813176149064&query=%E8%9B%8B%E7%99%BD%E9%85%B6&lattr=&xzhid=42454205&pi=b2b.shop.main.10..6097468723170060&ca tegory=%E5%8C%96%E5%B7%A5%E8%83%BD%E6%BA%90%3B%E9%A3%9F%E5%93%81%E6%B7%BB%E5%8A%A0%E5%89%82%3B%E9%85%B6%E5%88%B6% E5%89%82&fid=67436544%2C1735622905720&iid=15410f6f494316e1221d546ef3233cc4&miniId=8469&jid=2930774346&prod_type=0.

[0023] Example 1 A rose fermentation product comprises the following raw materials in parts by weight: 1000 g of rose flower, 300 g of rose leaf, 100 g of rose root, 100 g of red peony root, 100 g of trehalose and 100 g of oatmeal powder.

[0024] The preparation method of rose fermentation product comprises the following steps: (1) Pretreatment: Weigh 1000 g of rose flower, 300 g of rose leaf, 100 g of rose root, 100 g of red peony root, 100 g of trehalose, and 100 g of oatmeal powder according to the proportion, grind them, pass through a 60-mesh sieve, add them to 20 kg of pure water, and add 5 g of pectinase (5 × 10 4 U / g and 5g papain (5×10 4 U / g), treated at 50℃ for 30min under stirring conditions; sterilized at 105℃ for 15min to obtain rose culture medium; (2) Preliminary fermentation: ferment the Lactobacillus rhamnosus seed solution (bacterial concentration 10 6 CFU / ml) was inoculated into rose culture medium, and cultured in an anaerobic incubator at 37°C for 12 h. 10% sodium hydroxide solution was added to adjust the pH to 6.0; (3) Secondary fermentation: Bifidobacterium longum: Lactobacillus acidophilus: Lactobacillus plantarum: Kluyveromyces marxianus = 2:1:1:2 (mass ratio, bacterial concentration 1×10 8 CFU / ml) was inoculated into the preliminary fermentation liquid, and after culturing at 37℃ in an anaerobic incubator for 24h, the temperature was adjusted to 28-31℃ and fermented for 48h; (4) Decolorization: add 10 g of activated carbon, heat to 80 °C, stir for 10 min, centrifuge at 10,000 rpm to obtain a centrifuge, filter with a 10 μm filter membrane to obtain a light yellow clear filtrate; (5) Concentration treatment: The clarified filtrate is concentrated to 50% of the original volume by vacuum rotary evaporation equipment to obtain the rose fermentation product. Figure 1 shown.

[0025] After testing, the rose polysaccharide content in the rose fermentation product was 10.53g / L, and the polyphenol content was 5.80g / L.

[0026] Example 2 A rose fermentation product comprises the following raw materials in parts by weight: 1000 g of rose flower, 300 g of rose leaf, 100 g of rose root, 100 g of red peony root, 100 g of trehalose and 100 g of oatmeal powder.

[0027] The preparation method of rose fermentation product comprises the following steps: (1) Pretreatment: Weigh 1000 g of rose flower, 300 g of rose leaf, 100 g of rose root, 100 g of red peony root, 100 g of trehalose, and 100 g of oatmeal powder according to the proportion, grind them, pass through a 100-mesh sieve, add them to 20 kg of pure water, and add 5 g of pectinase (5 × 10 4 U / g and 5g papain (5×10 4 U / g), treated at 50℃ for 30min under stirring conditions; sterilized at 105℃ for 15min to obtain rose culture medium; (2) Preliminary fermentation: ferment the Lactobacillus rhamnosus seed solution (bacterial concentration 10 6 CFU / ml) was inoculated into rose culture medium, and cultured in an anaerobic incubator at 37°C for 14 h. 10% sodium hydroxide solution was added to adjust the pH to 6.0; (3) Secondary fermentation: Bifidobacterium longum: Lactobacillus acidophilus: Lactobacillus plantarum: Kluyveromyces marxianus = 2:1:1:2 (mass ratio, bacterial concentration 2×10 7 CFU / ml) was inoculated into the preliminary fermentation liquid, and after culturing at 37℃ in an anaerobic incubator for 24h, the temperature was adjusted to 28-31℃ and fermented for 72h; (4) Decolorization: add 8 g of activated carbon, heat to 80 °C, stir for 10 min, centrifuge at 10,000 rpm to obtain the centrifuge liquid, filter with a 10 μm filter membrane to obtain a light yellow clear filtrate; (5) Concentration treatment: The clarified filtrate is concentrated to 50% of the original volume by a vacuum rotary evaporation device to obtain a rose fermentation product.

[0028] After testing, the rose polysaccharide content in the rose fermentation product was 10.46g / L, and the polyphenol content was 5.83g / L.

[0029] Example 3 A rose fermentation product comprises the following raw materials in parts by weight: 1000 g of rose flower, 300 g of rose leaf, 100 g of rose root, 100 g of red peony root, 100 g of trehalose and 100 g of oatmeal powder.

[0030] The preparation method of rose fermentation product comprises the following steps: (1) Pretreatment: Weigh 1000 g of rose flower, 300 g of rose leaf, 100 g of rose root, 100 g of red peony root, 100 g of trehalose, and 100 g of oatmeal powder according to the proportion, grind them, pass through a 100-mesh sieve, add them to 20 kg of pure water, and add 6 g of pectinase (5 × 10 4 U / g and 5g papain (5×10 4 U / g), treated at 50℃ for 30min under stirring conditions; sterilized at 105℃ for 15min to obtain rose culture medium; (2) Preliminary fermentation: ferment the Lactobacillus rhamnosus seed solution (bacterial concentration 10 6 CFU / ml) was inoculated into rose culture medium, and cultured in an anaerobic incubator at 37°C for 16 h. 10% sodium hydroxide solution was added to adjust the pH to 6.0; (3) Secondary fermentation: Bifidobacterium longum: Lactobacillus acidophilus: Lactobacillus plantarum: Kluyveromyces marxianus = 2:1:1:2 (mass ratio, bacterial concentration 8×10 6 CFU / ml) was inoculated into the preliminary fermentation liquid, and after culturing at 37℃ in an anaerobic incubator for 24h, the temperature was adjusted to 28-31℃ and fermented for 96h; (4) Decolorization: add 10 g of activated carbon, heat to 80 °C, stir for 10 min, centrifuge at 10,000 rpm to obtain a centrifuge, filter with a 10 μm filter membrane to obtain a light yellow clear filtrate; (5) Concentration treatment: The clarified filtrate is concentrated to 50% of the original volume by a vacuum rotary evaporation device to obtain a rose fermentation product.

[0031] After testing, the rose polysaccharide content in the rose fermentation product was 10.41g / L, and the polyphenol content was 5.81g / L.

[0032] Example 4 A method for preparing rose polysaccharide, comprising the following steps: Take 5 kg of the rose fermentation product prepared according to the method of Example 1, add 50 g of composite protease, adjust the pH to 7, and treat at 37°C for 2 h. After digestion, treat at 80°C for 2 min. Add to 500 g of 95% ethanol, let stand for 12 h, centrifuge at 12000 r / min for 10 min, obtain a precipitate, and dry at 50°C to obtain rose polysaccharide.

[0033] After testing, the content of rose polysaccharide in the sample was 93.5%, and the rest was impurities. The polysaccharide had good biological activity.

[0034] Example 5 A method for preparing rose polysaccharide, comprising the following steps: Take 5 kg of rose fermentation product prepared according to the method of Example 2, add 50 g of composite protease, adjust the pH to 7, and treat at 37°C for 2 h. After digestion, treat at 80°C for 2 min. Add to 500 g of 95% ethanol, let stand for 12 h, centrifuge at 12000 r / min for 10 min, obtain precipitate, and dry at 50°C to obtain rose polysaccharide.

[0035] After testing, the content of rose polysaccharide in this sample was 91.2%.

[0036] Example 6 A method for preparing rose polysaccharide, comprising the following steps: Take 5 kg of the rose fermentation product prepared according to the method of Example 3, add 50 g of composite protease, adjust the pH to 7, and treat at 37°C for 2 h. After digestion, treat at 80°C for 2 min. Add to 500 g of 95% ethanol, let stand for 12 h, centrifuge at 12000 r / min for 10 min, obtain a precipitate, and dry at 50°C to obtain rose polysaccharide.

[0037] After testing, the content of rose polysaccharide in this sample was 90.1%.

[0038] Comparative Example 1 A method for preparing rose polysaccharide comprises the following raw materials in parts by weight: 1000 g of rose flowers.

[0039] The specific steps are as follows: (1) After the rose flowers were crushed, 20 L of deionized water was added, ultrasonicated for 10 min, and then heated in 90 °C water for 2 h, and filtered to obtain the extract; the filter residue was repeatedly extracted twice with 10 L × 2 deionized water, the extracts were combined, and concentrated to a residual volume of 20 L; the rose polysaccharide content in the extract was detected to be 5.72 g / L, and the polyphenol content was 2.40 g / L.

[0040] (2) Add 60 L of anhydrous ethanol to the concentrated extract, let it stand for 12 hours, centrifuge, and dry the solid at 50°C to obtain rose polysaccharide.

[0041] After testing, the content of rose polysaccharide in this sample was 74.9%.

[0042] Comparative Example 2 A method for preparing rose polysaccharide comprises the following raw materials in parts by weight: 1000 g of rose flowers.

[0043] (1) After crushing the rose, add 20L of deionized water and sonicate for 10 minutes; then add (2) Crushing of roses: Add roses to 20L of deionized water and soak for 30min, heat to 85℃ and soak for 2h; then transfer the liquid containing roses to a colloid mill for crushing to obtain rose crushing liquid; (3) Heat the rose flower liquid for disinfection and keep it for 10 minutes; then cool it to 30°C, add 0.03% brewer's yeast according to the total weight of the liquid, ferment it at 28°C for 48 hours, and when the alcohol content reaches 5 degrees, heat it to 32°C, add 0.05% acetic acid bacteria, ferment it at 35°C for 48 hours, then add 0.3% plant lactobacillus at 35°C, and ferment it at 35°C for 72 hours. After the fermentation is completed, heat it to 85-90°C, keep it warm for 5 minutes, and terminate the fermentation; the rose polysaccharide content in the fermentation liquid is 5.11g / L, and the polyphenol content is 2.23g / L (4) The fermentation liquid is filtered using a filter membrane to obtain a clear liquid; 60 L of anhydrous ethanol is added to the clear liquid, the mixture is allowed to stand for 12 h, and the mixture is centrifuged. The solid is dried at 50 °C to obtain rose polysaccharide.

[0044] After testing, the content of rose polysaccharide in this sample was 58.2%.

[0045] By comparing Examples 1 to 6 and Comparative Examples 1 to 2, it can be seen that the contents of rose polysaccharides and polyphenols in the rose fermentation product prepared by the present invention are significantly increased, wherein the content of rose polysaccharides is increased by 84.1%, and the content of polyphenols is increased by 141.6%.

[0046] Test Case 1. Whitening efficacy test In the biosynthesis of melanin in the skin, tyrosinase is a key enzyme that acts on DOPA to form DOPA quinone, which spontaneously undergoes a series of reactions to finally form melanin. In a phosphoric acid solution with a pH of 6.8, tyrosinase can catalyze the conversion of DOPA into DOPA quinone, and the absorbance value can be measured at 475nm on a spectrophotometer. Raw materials that inhibit tyrosinase activity can reduce the conversion of DOPA into DOPA quinone, thereby reducing the absorbance value. The inhibitory effect of the raw material on tyrosinase activity can be evaluated based on the change in absorbance value.

[0047] Referring to Table 1, use 10mL test tubes to set up sample tubes (T), sample background (T0), enzyme reaction tubes (C) and solvent background (C0). For each sample tube (T) of each tested concentration, three parallel tubes need to be set up, and three parallel tubes need to be set up for the enzyme reaction tube (C). 1mL of sample solution of the same concentration is added to the sample tube (T) and sample background (T0), and 1m sodium dihydrogen phosphate-citrate buffer is added to the enzyme reaction tube (C) and solvent background (C0).

[0048] Add 0.5 mL of tyrosinase solution to each of the sample tube (T) and the enzyme reaction tube (C). Replace the sample background (T0) and the solvent background (C0) with 0.5 ml of disodium hydrogen phosphate-citrate buffer. Mix the sample and tyrosinase thoroughly and incubate in a 37°C water bath for 10 minutes.

[0049] 2 mL of L-DOPA solution was added to each tube in turn, and the reaction time of each tube was controlled to be 5 minutes. The reaction solution of each tube was immediately transferred to a cuvette, and the absorbance value was measured at 475 nm.

[0050] Table 1 - Product addition amount

[0051] The preparation method of the rose fermentation product sample is: dilute the rose fermentation product with pure water 50 times. The preparation method of the rose polysaccharide sample is: dilute the rose polysaccharide with pure water 1000 times.

[0052] Tyrosinase activity inhibition rate (%) = (1-(A T -A T0 )÷(A C -A C0 ))×100%; In the formula, A T -The absorbance value of the sample tube, i.e., the absorbance value of the solution after the sample reacts with tyrosinase; A T0 -Sample background absorbance; A C - The average value of the absorbance of the enzyme reaction tube three times, that is, the absorbance of the reaction between tyrosinase and DOPA when no sample is added; A C0 -Solvent background absorbance.

[0053] The test results are shown in Table 2.

[0054] Table 2 - Absorbance calculation results

[0055] It can be seen from the absorbance test results in Table 2 that the rose fermentation product and rose polysaccharide prepared by the present invention have a good inhibitory effect on the activity of tyrosinase.

[0056] 2. Anti-inflammatory test Preparation of sample solution: Using lipopolysaccharide (10,000 units / mL) as the diluent, prepare the samples into 1% volume concentration solutions and filter them through a 0.22 μm filter membrane.

[0057] Control group: Raw264.7 cells were cultured at 5×10 4 / ml were inoculated in 24-well plates and cultured at 37°C, 5% CO2 for 24 h. Serum-free culture medium was added and cultured for another 24 h. Inflammatory factors were detected using ELISA kits.

[0058] Model group: Raw264.7 cells were cultured at 5×10 4 / ml was inoculated into a 24-well plate and cultured at 37°C, 5% CO2 for 24 h. An equal volume of lipopolysaccharide (10,000 units / mL) to that of the serum-free medium of the normal control group was added and cultured for another 24 h. Inflammatory factors were detected using an ELISA kit.

[0059] Experimental group: Raw264.7 cells were cultured at 5×10 4 / ml were inoculated in 24-well plates and cultured at 37°C, 5% CO2 for 24 h. The sample solution was added with an equal volume of serum-free culture medium to that of the normal control group and cultured for another 24 h. Inflammatory factors were detected using ELISA kits.

[0060] The inhibition rate was calculated as follows: ; Table 3 - Inflammation inhibition rate

[0061] It can be seen from the data in Table 3 that after being treated with the sample solution, the inflammatory factors decreased significantly, rose fermentation products and rose polysaccharides had a significant inhibitory effect on inflammation, and the anti-inflammatory effect of rose fermentation products was more obvious.

[0062] 3. Skin elasticity test efficacy 60 volunteers (half male and half female, aged 20-35) were selected. Before the test, the skin elasticity condition A0 of the left face was tested with a skin elasticity tester (Cutometer MPA 580). The volunteers used the samples of the embodiment (the samples were (1) an essence containing 2% of rose fermentation product and (2) an essence containing 0.1% of rose polysaccharide) on the left face respectively according to the group, and applied them to the face twice a day in the morning and evening for 2 weeks. The skin elasticity condition A1 of the left face was tested with a skin elasticity tester (Cutometer MPA 580), and the skin elastin growth rate was calculated, skin elasticity growth rate=(A1-A0) / A0. The test results are shown in Table 4.

[0063] Table 4 - Skin elastin test results

[0064] The results of the skin elasticity test showed that 2% rose fermentation products can significantly improve the elasticity of volunteers' facial skin, with an increase rate of more than 60%. 0.1% fermentation product polysaccharide can improve the elasticity of volunteers' facial skin by more than 68%. Both have good anti-aging effects.

[0065] Although the present invention has been described in detail with reference to the accompanying drawings and in combination with the preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily think of changes or substitutions within the technical scope disclosed by the present invention, and these shall be within the scope of protection of the present invention.

Claims

1. A method for preparing a rose fermentation product, characterized in that: The following steps are involved: (1) Pretreatment: Weigh rose flowers, rose leaves, rose roots, red peony root, trehalose and oatmeal powder in proportion, crush, sieve, add to pure water, add pectinase and papain, and treat at 40-50°C for 30 min under stirring; sterilize at 105°C for 15-20 min to obtain rose culture medium; (2) Preliminary fermentation: inoculate the fermented Lactobacillus rhamnosus seed liquid into rose culture medium and culture it in an anaerobic incubator at 37°C for 12 to 16 hours; add 10% sodium hydroxide solution to adjust the pH to 6.0 to obtain the preliminary fermentation liquid; (3) Secondary fermentation: Inoculate the composite bacterial liquid of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus plantarum and Kluyveromyces marxianus into the primary fermentation liquid, incubate in an anaerobic incubator at 37°C for 24 hours, then adjust the temperature to 28-31°C and ferment for another 48-96 hours; (4) Decolorization: add 0.5% to 1.0% activated carbon based on the weight of the fermentation liquid, heat to 80°C, stir for 10 min, centrifuge at 10,000 rpm to obtain a centrifuge, filter with an 8-10 μm filter membrane to obtain a light yellow clear filtrate; (5) Concentration treatment: The clarified filtrate is concentrated to 50% by vacuum rotary evaporation equipment to obtain the rose fermentation product.

2. The method for preparing a rose fermentation product according to claim 1, characterized in that: In step (1), the following raw materials are included in weight parts: 10 parts of rose flowers, 3 parts of rose leaves, 1 part of rose roots, 1 part of red peony root, 1 part of trehalose, 1 part of oatmeal powder, and 200 parts of water.

3. The method for preparing the rose fermentation product according to claim 1, characterized in that: In step (1), the amount of pectinase added is 0.5% to 0.6% based on the total weight of the raw materials; the enzyme activity is ≥5×10 4 U / g; the amount of papain added is 0.4%~0.6% based on the total weight of the raw materials, and the enzyme activity is ≥5×10 4 U / g.

4. The method for preparing the rose fermentation product according to claim 1, characterized in that: The bacterial concentration in the Lactobacillus rhamnosus fermentation seed liquid in step (2) is ≥10 6 CFU / ml.

5. The method for preparing a rose fermentation product according to claim 1, characterized in that: In step (2), the amount of Lactobacillus rhamnosus fermented seed liquid added is 3% to 5% by weight of the rose culture medium.

6. The method for preparing the rose fermentation product according to claim 1, characterized in that: In the composite bacterial solution of step (3), the mass ratio of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus plantarum and Kluyveromyces marxianus is 2:1:1:2; the bacterial concentration is 10 6 ~10 8 CFU / ml.

7. A rose fermentation product prepared by the preparation method according to claim 1, characterized in that: The rose polysaccharide content in the rose fermentation product is ≥10.4g / L, and the polyphenol content is ≥5.8g / L.

8. A method for extracting rose polysaccharides, characterized in that: The steps are as follows: add 1 part of composite protease to 100 parts of the rose fermentation product as claimed in claim 7, adjust the pH to 7-8; treat at 37°C for 2-4 hours; after digestion, treat at 80°C for 2 minutes; add to 3-4 parts of 95% ethanol, let stand for 12-24 hours, centrifuge at 12000-16000 r / min for 5-10 minutes, obtain precipitation, and dry at 45-60°C to obtain rose polysaccharide.

9. Use of the rose fermentation product as claimed in claim 7 in the preparation of whitening and anti-inflammatory products.

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