Preparation method of rice fermentation product filtrate for relieving and controlling oil as well as product and application thereof

Abstract

The invention provides a preparation method of rice fermentation product filtrate for relieving and controlling oil as well as a product and application thereof, and the preparation method comprises the following steps: (1) mixing rice and water, heating, and carrying out gelatinization reaction; (2) adjusting the pH value, and adding saccharifying enzyme and neutral protease to carry out enzymolysis reaction; and (3) performing enzyme deactivation, inoculating compound bacteria for fermentation, performing sterilization after fermentation, and performing solid-liquid separation to obtain the rice fermentation product filtrate. According to the preparation method disclosed by the invention, the water solubility, absorptivity and stability of the rice fermentation product filtrate are improved, and the prepared rice fermentation product filtrate can remarkably inhibit the activity of 5alpha-reductase and inhibit abnormal secretion of skin grease from the source; meanwhile, the activity of hyaluronidase and tyrosinase is remarkably inhibited, the DPPH free radical scavenging rate is increased, the excellent effects of relieving, controlling oil, resisting oxidation, whitening and moisturizing are achieved, and the skin is endowed with refreshing and pleasant skin feeling experience while the skin barrier is repaired.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological fermentation, and particularly relates to a preparation method, product and application of a soothing and oil-control rice fermentation product filtrate. Background Art

[0002] The rice fermentation product filtrate has been applied in the fields of medicine, food and daily chemical industry. However, due to the instability of its oil-control performance, its effect in actual application is limited.

[0003] CN119700633A discloses a preparation method of a cosmetic raw material derived from a rice fermentation product filtrate. The specific operation process is to filter the rice fermentation product to remove solid impurities, further filter and clarify, adjust the pH of the solution with alkali, then filter to remove colored substances, add an adsorbent for decolorization, filter to remove the adsorbent, and sterilize and preserve to obtain a rice fermentation product active substance solution, which is used as a cosmetic raw material. The production process does not involve the use of organic solvents, has a high content of active substances, produces less waste, and has a stable process, which is suitable for large-scale production.

[0004] CN105193658A discloses a rice fermentation raw pulp cosmetic and its preparation method and application. The method for preparing the rice fermentation raw pulp includes the following steps: fermenting and culturing an initial system composed of Bifidobacterium bifidum, rice dry powder and water. The prepared rice fermentation raw pulp does not contain any chemical components and can be directly used as a finished product of a facial mask, essence or toner, which is more natural than other existing products on the market and will not cause any side effects to the skin.

[0005] The composition of the rice fermentation product filtrate prepared by the prior art still has limitations, and it cannot effectively extract and utilize the active components in rice. Therefore, there is an urgent need to develop a rice fermentation product filtrate with high water solubility and absorbability. Summary of the Invention

[0006] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a preparation method, product and application of a soothing and oil-control rice fermentation product filtrate. The preparation method of the present invention improves the water solubility and absorbability of the rice fermentation product filtrate, and the prepared rice fermentation product filtrate can exert excellent soothing and oil-control, antioxidant and whitening and moisturizing effects.

[0007] To achieve the purpose of this invention, the following technical solutions are adopted by the present invention:

[0008] In the first aspect, the present invention provides a preparation method of a soothing and oil-control rice fermentation product filtrate, and the preparation method includes:

[0009] (1) Mix rice and water and heat them to carry out a gelatinization reaction;

[0010] (2) Adjust the pH value, add glucoamylase and neutral protease, and carry out an enzymatic hydrolysis reaction;

[0011] (3) Inactivate the enzyme, inoculate with a complex bacterium for fermentation, sterilize after completion, and perform solid-liquid separation to obtain the filtrate of the rice fermentation product.

[0012] The preparation method of the present invention improves the water solubility, absorbability and stability of the filtrate of the rice fermentation product. The prepared filtrate of the rice fermentation product can significantly inhibit the activity of 5α-reductase, inhibit the abnormal secretion of skin oil from the root, and endow the skin with a fresh and pleasant skin feel experience; at the same time, it significantly inhibits the activities of hyaluronidase and tyrosinase, has a high DPPH free radical scavenging rate, and exerts excellent functions of soothing oil control, antioxidant and whitening and moisturizing.

[0013] The present invention creatively uses glucoamylase and neutral protease in the preparation of the filtrate of the rice fermentation product. The two have a synergistic effect and can fully decompose macromolecular substances in rice into various small molecule active components such as active peptides, amino acids, polysaccharides and organic acids, providing an abundant substrate source for the fermentation of the complex bacterium.

[0014] Preferably, in step (1), the mass ratio of the rice to water is 1:(40 - 50).

[0015] The specific point values in (40 - 50) can be 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50, etc.

[0016] Preferably, the heating temperature is 80 - 90°C, for example, it can be 80°C, 81°C, 82°C, 83°C, 84°C, 85°C, 86°C, 87°C, 88°C, 89°C or 90°C, etc.; the time is 50 - 70 min, for example, it can be 50 min, 52 min, 54 min, 56 min, 58 min, 60 min, 62 min, 64 min, 66 min, 68 min or 70 min, etc.

[0017] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0018] Preferably, in step (2), the pH value is 5.5 - 6.5, for example, it can be 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4 or 6.5, etc.

[0019] Preferably, the mass percentage content of glucoamylase and neutral protease in the enzymatic hydrolysis reaction system is independently 0.01 - 1%, for example, it can be 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%, etc.

[0020] Preferably, the temperature of the enzymatic hydrolysis reaction is 40-60°C, for example, it can be 40°C, 42°C, 44°C, 46°C, 48°C, 50°C, 52°C, 54°C, 56°C, 58°C or 60°C, etc.; the time is 2-4h, for example, it can be 2h, 2.2h, 2.4h, 2.6h, 2.8h, 3h, 3.2h, 3.4h, 3.6h, 3.8h or 4h, etc.

[0021] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0022] Preferably, after the enzymatic hydrolysis reaction, it further includes an ultrasonic step.

[0023] Preferably, the frequency of the ultrasonic wave is 20-40kHz, for example, it can be 20kHz, 22kHz, 24kHz, 26kHz, 28kHz, 30kHz, 32kHz, 34kHz, 36kHz, 38kHz or 40kHz, etc.; the time is 10-30min, for example, it can be 10min, 12min, 14min, 16min, 18min, 20min, 22min, 24min, 26min, 28min or 30min, etc.

[0024] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0025] In the present invention, ultrasonic treatment at a specific frequency is carried out after the enzymatic hydrolysis reaction. Through cavitation, the enzymatic hydrolysis products are further decomposed and transformed, improving the conversion and utilization rate of the composite bacteria for the substrate, and enhancing the soothing oil control, antioxidant, and whitening and moisturizing effects of the filtrate of the rice fermentation product.

[0026] Preferably, in step (3), the composite bacteria include Saccharomyces cerevisiae and Lactobacillus acidophilus.

[0027] The present invention creatively uses Saccharomyces cerevisiae and Lactobacillus acidophilus in the preparation of the filtrate of the rice fermentation product. The two have a synergistic effect, carrying out composite fermentation on the enzymatic hydrolysis products, producing various amino acids, polypeptides, vitamins, organic acids and minerals, and the obtained filtrate of the rice fermentation product has excellent soothing oil control, antioxidant, and whitening and moisturizing effects.

[0028] Preferably, the viable count ratio of Saccharomyces cerevisiae to Lactobacillus acidophilus is (1-5):(1-5).

[0029] The specific point values in the first (1-5) can be 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5, etc.

[0030] The specific point values in the second (1-5) can be 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, etc.

[0031] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0032] Preferably, the fermentation includes two fermentations.

[0033] Preferably, the inoculation amount of the composite bacteria in the first fermentation is 5-10%, for example, it can be 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10%, etc.

[0034] Preferably, the temperature of the first fermentation is 25-35°C, for example, it can be 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C or 35°C, etc.; the time is 8-12h, for example, it can be 8h, 8.5h, 9h, 9.5h, 10h, 10.5h, 11h, 11.5h or 12h, etc.

[0035] Preferably, the inoculation amount of the composite bacteria in the second fermentation is 2-5%, for example, it can be 2%, 2.2%, 2.5%, 2.8%, 3%, 3.2%, 3.5%, 3.8%, 4%, 4.2%, 4.5%, 4.8% or 5%, etc.

[0036] Preferably, the temperature of the second fermentation is 28-30°C, for example, it can be 28°C, 28.2°C, 28.4°C, 28.6°C, 28.8°C, 29°C, 29.2°C, 29.4°C, 29.6°C, 29.8°C or 30°C, etc.; the time is 24-48h, for example, it can be 24h, 26h, 28h, 30h, 32h, 34h, 36h, 38h, 40h, 42h, 44h, 46h or 48h, etc.

[0037] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0038] In the present invention, the composite bacteria carry out a specific two-step fermentation process, which can further enhance the soothing oil control, antioxidant and whitening and moisturizing effects of the filtrate of the rice fermentation product. The composite bacteria carry out the first fermentation based on the enzymatic hydrolysis product, and initially carry out metabolic transformation; based on the metabolic product of the first fermentation, the composite bacteria carry out the second fermentation to further produce specific active ingredients beneficial to the skin, optimizing the active composition and its stability of the filtrate of the rice fermentation product.

[0039] Preferably, in step (3), the temperature for enzyme inactivation is 90 - 100°C, for example, it can be 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C or 100°C, etc.; the time is 25 - 40 min, for example, it can be 25 min, 26 min, 27 min, 28 min, 29 min, 30 min, 31 min, 32 min, 33 min, 34 min, 35 min, 36 min, 37 min, 38 min, 39 min or 40 min, etc.

[0040] Preferably, the temperature for sterilization is 90 - 100°C, for example, it can be 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C or 100°C, etc.; the time is 25 - 40 min, for example, it can be 25 min, 26 min, 27 min, 28 min, 29 min, 30 min, 31 min, 32 min, 33 min, 34 min, 35 min, 36 min, 37 min, 38 min, 39 min or 40 min, etc.

[0041] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0042] Preferably, in step (3), the method of solid - liquid separation is centrifugation.

[0043] Preferably, the rotation speed of centrifugation is 8000 - 10000 rpm, for example, it can be 8000 rpm, 8200 rpm, 8400 rpm, 8600 rpm, 8800 rpm, 9000 rpm, 9200 rpm, 9400 rpm, 9600 rpm, 9800 rpm or 10000 rpm, etc.; the time is 10 - 15 min, for example, it can be 10 min, 10.5 min, 11 min, 11.5 min, 12 min, 12.5 min, 13 min, 13.5 min, 14 min, 14.5 min or 15 min, etc.

[0044] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0045] Preferably, after the solid - liquid separation, it further includes the step of ultrafiltration membrane filtration.

[0046] In the second aspect, the present invention provides a filtrate of rice fermentation product prepared by the preparation method as described in the first aspect.

[0047] The filtrate of rice fermentation product provided by the present invention has good water - solubility, stability and absorbability, and exhibits excellent effects of soothing oil control, antioxidant and whitening and moisturizing.

[0048] In a third aspect, the present invention provides the use of the filtrate of the rice fermentation product as described in the second aspect in cosmetics.

[0049] The filtrate of the rice fermentation product provided by the present invention has good water solubility, stability and absorbability, and exerts excellent soothing oil control, antioxidant and whitening and moisturizing effects. It can be widely used in cosmetics, significantly reducing the addition amount of surfactants, fragrances or preservatives, reducing the irritation to the skin, and enhancing the mildness of cosmetic products.

[0050] Preferably, the cosmetic includes any one of lotion, emulsion, cream, essence, spray, facial mask, sunscreen, shampoo, body wash, hair conditioner or hair mask.

[0051] The filtrate of the rice fermentation product provided by the present invention can not only be applied to skin care products, but also to washing and care products, improving the fineness and durability of the foam of surfactants, ensuring mildness and no irritation while having excellent oil control and cleaning effects, and giving the skin a fresh and pleasant skin feeling experience.

[0052] In a fourth aspect, the present invention provides the use of the filtrate of the rice fermentation product as described in the second aspect in the preparation of a 5α-reductase inhibitor.

[0053] The filtrate of the rice fermentation product provided by the present invention can significantly inhibit the activity of 5α-reductase, and inhibit the abnormal secretion of skin oil from the root.

[0054] Compared with the prior art, the present invention has the following beneficial effects:

[0055] The preparation method of the present invention improves the water solubility, absorbability and stability of the filtrate of the rice fermentation product. The prepared filtrate of the rice fermentation product can significantly inhibit the activity of 5α-reductase, inhibit the abnormal secretion of skin oil from the root, and give the skin a fresh and pleasant skin feeling experience; at the same time, it significantly inhibits the activities of hyaluronidase and tyrosinase, has a high DPPH free radical scavenging rate, and exerts excellent soothing oil control, antioxidant and whitening and moisturizing effects. Detailed Embodiments

[0056] To further elaborate on the technical means and effects adopted by the present invention, the following further illustrates the technical solutions of the present invention in combination with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0057] For those not specifying specific techniques or conditions in the embodiments, they shall be carried out according to the techniques or conditions described in the literature in the field or according to the product specifications. For those reagents or instruments not indicating the manufacturer, they are all conventional products that can be obtained through regular channels.

[0058] The sources of the materials used in the following specific embodiments are as follows:

[0059] Raw material name Purchasing manufacturer Model Glucoamylase Shandong Sukahan Bioengineering Co., Ltd. Glucoamylase Neutral protease Shandong Sukahan Bioengineering Co., Ltd. Neutral protease Ferulic acid esterase Beijing Pusitang Biotechnology Co., Ltd. Ferulic acid esterase α-Amylase Shandong Sukahan Bioengineering Co., Ltd. α-Amylase Saccharomyces cerevisiae American Type Culture Collection ATCC 26603 Lactobacillus acidophilus American Type Culture Collection ATCC 4356

[0060] Example 1

[0061] This example provides a method for preparing a soothing and oil-control rice fermentation product filtrate. The preparation method includes:

[0062] (1) After pulverizing and sieving rice, mix it with water at a mass ratio of 1:45, heat at 85 °C for 60 min, and carry out a gelatinization reaction;

[0063] (2) Adjust the pH value to 6, add 0.5% saccharifying enzyme and 0.4% neutral protease by mass percentage, and carry out enzymatic hydrolysis at 50 °C for 3 h; after the enzymatic hydrolysis reaction ends, carry out ultrasonic treatment at a frequency of 30 kHz for 20 min;

[0064] (3) Maintain at 95 °C for 30 min to inactivate the enzyme; inoculate 4% activated yeast liquid and 4% activated Lactobacillus acidophilus liquid (the concentration of both liquid cultures is 1×10 10 CFU / mL), carry out the first fermentation, and ferment at 30 °C for 10 h; then inoculate 3% activated yeast liquid and 1% activated Lactobacillus acidophilus liquid (the concentration of both liquid cultures is 1×10 10 CFU / mL), carry out the second fermentation, and ferment at 29 °C for 36 h; after the fermentation ends, maintain at 95 °C for 30 min to sterilize; centrifuge at 9000 rpm for 12 min, collect the liquid, and filter it through an ultrafiltration membrane to obtain the rice fermentation product filtrate.

[0065] Example 2

[0066] This example provides a method for preparing a soothing and oil-control rice fermentation product filtrate. The preparation method includes:

[0067] (1) After pulverizing and sieving rice, mix it with water at a mass ratio of 1:40, heat at 80 °C for 70 min, and carry out a gelatinization reaction;

[0068] (2) Adjust the pH value to 5.5, add 0.8% saccharifying enzyme and 0.1% neutral protease by mass percentage, and carry out enzymatic hydrolysis at 40 °C for 4 h; after the enzymatic hydrolysis reaction ends, carry out ultrasonic treatment at a frequency of 20 kHz for 30 min;

[0069] (3) Maintain at 90 °C for 40 min to inactivate the enzyme; inoculate 2% activated yeast liquid and 3% activated Lactobacillus acidophilus liquid (the concentration of both liquid cultures is 1×10 10 CFU / mL), carry out the first fermentation, and ferment at 25 °C for 12 h; then inoculate 2% activated yeast liquid and 3% activated Lactobacillus acidophilus liquid (the concentration of both liquid cultures is 1×10 10CFU / mL), perform the second fermentation at 28°C for 48 h; after the fermentation is completed, maintain at 90°C for 40 min for sterilization; centrifuge at 8000 rpm for 15 min, collect the liquid, and filter through an ultrafiltration membrane to obtain the filtrate of the rice fermentation product.

[0070] Example 3

[0071] This example provides a method for preparing a filtrate of a rice fermentation product with soothing and oil-control effects. The preparation method includes:

[0072] (1) After pulverizing and sieving the rice, mix it with water at a mass ratio of 1:50, and heat at 90°C for 50 min for gelatinization reaction;

[0073] (2) Adjust the pH value to 6.5, add 0.1% glucoamylase and 0.8% neutral protease by mass percentage, and enzymatically hydrolyze at 60°C for 2 h; after the enzymatic hydrolysis reaction is completed, ultrasonicate at a frequency of 40 kHz for 10 min;

[0074] (3) Maintain at 100°C for 25 min to inactivate the enzymes; inoculate 6% activated liquid of Saccharomyces cerevisiae and 4% activated liquid of Lactobacillus acidophilus (the concentration of both bacterial liquids is 1×10 10 CFU / mL), perform the first fermentation at 35°C for 8 h; then inoculate 1% activated liquid of Saccharomyces cerevisiae and 1% activated liquid of Lactobacillus acidophilus (the concentration of both bacterial liquids is 1×10 10 CFU / mL), perform the second fermentation at 30°C for 24 h; after the fermentation is completed, maintain at 100°C for 25 min for sterilization; centrifuge at 10000 rpm for 10 min, collect the liquid, and filter through an ultrafiltration membrane to obtain the filtrate of the rice fermentation product.

[0075] Example 4

[0076] This example provides a method for preparing a filtrate of a rice fermentation product with soothing and oil-control effects. The difference from Example 1 is only that: in step (3), the activated liquid of Saccharomyces cerevisiae is not added, and the reduced amount is supplemented by the activated liquid of Lactobacillus acidophilus, and the other steps remain unchanged.

[0077] Example 5

[0078] This example provides a method for preparing a filtrate of a rice fermentation product with soothing and oil-control effects. The difference from Example 1 is only that: in step (3), the activated liquid of Lactobacillus acidophilus is not added, and the reduced amount is supplemented by the activated liquid of Saccharomyces cerevisiae, and the other steps remain unchanged.

[0079] Example 6

[0080] This embodiment provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Embodiment 1 in that: in step (3), replace "inoculate 4% of the activated liquid of Saccharomyces cerevisiae and 4% of the activated liquid of Lactobacillus acidophilus (the concentration of both liquid cultures is 1×10 10 CFU / mL), conduct the first fermentation, and ferment at 30°C for 10 h; then inoculate 3% of the activated liquid of Saccharomyces cerevisiae and 1% of the activated liquid of Lactobacillus acidophilus (the concentration of both liquid cultures is 1×10 10 CFU / mL), conduct the second fermentation, and ferment at 29°C for 36 h" with "inoculate 7% of the activated liquid of Saccharomyces cerevisiae (the concentration of the liquid culture is 1×10 10 CFU / mL), conduct the first fermentation, and ferment at 30°C for 10 h; then inoculate 5% of the activated liquid of Lactobacillus acidophilus (the concentration of the liquid culture is 1×10 10 CFU / mL), conduct the second fermentation, and ferment at 29°C for 36 h", and keep other steps unchanged.

[0081] Example 7

[0082] This embodiment provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Embodiment 1 in that: in step (3), replace "inoculate 4% of the activated liquid of Saccharomyces cerevisiae and 4% of the activated liquid of Lactobacillus acidophilus (the concentration of both liquid cultures is 1×10 10 CFU / mL), conduct the first fermentation, and ferment at 30°C for 10 h; then inoculate 3% of the activated liquid of Saccharomyces cerevisiae and 1% of the activated liquid of Lactobacillus acidophilus (the concentration of both liquid cultures is 1×10 10 CFU / mL), conduct the second fermentation, and ferment at 29°C for 36 h" with "inoculate 5% of the activated liquid of Lactobacillus acidophilus (the concentration of the liquid culture is 1×10 10 CFU / mL), conduct the first fermentation, and ferment at 30°C for 10 h; then inoculate 7% of the activated liquid of Saccharomyces cerevisiae (the concentration of the liquid culture is 1×10 10 CFU / mL), conduct the second fermentation, and ferment at 29°C for 36 h", and keep other steps unchanged.

[0083] Example 8

[0084] This embodiment provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Embodiment 1 in that: in step (3), replace "inoculate 4% of the activated liquid of Saccharomyces cerevisiae and 4% of the activated liquid of Lactobacillus acidophilus (the concentration of both liquid cultures is 1×10 10 CFU / mL), conduct the first fermentation, and ferment at 30°C for 10 h; then inoculate 3% of the activated liquid of Saccharomyces cerevisiae and 1% of the activated liquid of Lactobacillus acidophilus (the concentration of both liquid cultures is 1×10 10"(CFU / mL), and perform the second fermentation at 29°C for 36 h" is replaced with "Inoculate 7% activated liquid of Saccharomyces cerevisiae and 5% activated liquid of Lactobacillus acidophilus (the concentration of both liquid cultures is 1×10 10 CFU / mL), and only perform one fermentation at 29°C for 46 h", and other steps remain unchanged.

[0085] Example 9

[0086] This example provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Example 1 in that: in step (2), "ultrasonic at a frequency of 30 kHz" is replaced with "ultrasonic at a frequency of 50 kHz", and other steps remain unchanged.

[0087] Example 10

[0088] This example provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Example 1 in that: in step (2), "ultrasonic at a frequency of 30 kHz" is replaced with "ultrasonic at a frequency of 10 kHz", and other steps remain unchanged.

[0089] Comparative Example 1

[0090] This comparative example provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Example 1 in that: in step (2), no glucoamylase is added, and the reduced amount is supplemented by neutral protease, and other steps remain unchanged.

[0091] Comparative Example 2

[0092] This comparative example provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Example 1 in that: in step (2), no neutral protease is added, and the reduced amount is supplemented by glucoamylase, and other steps remain unchanged.

[0093] Comparative Example 3

[0094] This comparative example provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Example 1 in that: in step (2), glucoamylase is replaced with an equal amount of ferulic acid esterase, and other steps remain unchanged.

[0095] Comparative Example 4

[0096] This comparative example provides a method for preparing a soothing and oil-control rice fermentation product filtrate, which is only different from Example 1 in that: in step (2), neutral protease is replaced with an equal amount of α-amylase, and other steps remain unchanged.

[0097] Comparative Example 5

[0098] This comparative example provides a method for preparing a soothing and oil-control rice fermentation product filtrate, and the preparation method includes:

[0099] (1) After crushing and sieving rice, mix it with water at a mass ratio of 1:45, heat at 85 °C for 60 min, and carry out a gelatinization reaction;

[0100] (2) Inoculate 4% activated yeast solution and 4% activated Lactobacillus acidophilus solution (the concentration of both solutions is 1×10 10 CFU / mL), carry out the first fermentation, and ferment at 30 °C for 10 h; then inoculate 3% activated yeast solution and 1% activated Lactobacillus acidophilus solution (the concentration of both solutions is 1×10 10 CFU / mL), carry out the second fermentation, and ferment at 29 °C for 36 h; after the fermentation ends, maintain at 95 °C for 30 min to carry out sterilization;

[0101] (3) Adjust the pH value to 6, add 0.5% glucoamylase and 0.4% neutral protease by mass percentage, carry out enzymatic hydrolysis at 50 °C for 3 h; after the enzymatic hydrolysis reaction ends, carry out ultrasonic treatment at a frequency of 30 kHz for 20 min; maintain at 95 °C for 30 min to inactivate the enzyme; centrifuge at 9000 rpm for 12 min, collect the liquid, and filter it through an ultrafiltration membrane to obtain the rice fermentation product filtrate.

[0102] Test Example 1 5α-Reductase Inhibition Rate

[0103] Prepare reagents: ① Testosterone solution: Dissolve testosterone in absolute ethanol to obtain a testosterone solution with a concentration of 5 mM; ② NADPH solution: Dissolve NADPH (reduced nicotinamide adenine dinucleotide phosphate) tetrasodium salt in water to obtain an NADPH solution with a concentration of 2 mM; ③ Reaction solution: Dissolve dithiothreitol (DTT) and sodium phosphate in water together to make the final concentrations of DTT and sodium phosphate 1 mM and 20 mM respectively to obtain the reaction solution; ④ Test sample solution: After freeze-drying the rice fermentation product filtrate prepared in Examples 1-10 or Comparative Examples 1-5, add water to prepare a solution with a final concentration of 10%.

[0104] Mix 155 μL of the reaction solution, 10 μL of the testosterone solution, 10 μL of the NADPH solution, 15 μL of the test sample solution, and 20 μL of 5α-reductase in sequence under the condition of 37 °C, continuously monitor the change in the absorbance value of the reaction system at 340 nm within 4 min, and reflect the activity of 5α-reductase with the change speed (in a reaction system at a temperature of 37 °C, the amount of enzyme that causes the NADPH concentration to decrease by 1 μM per minute is one enzyme activity unit), calculate the inactivated enzyme activity of 5α-reductase with reference to the NADPH standard curve, record the test result without the rice fermentation product filtrate as the initial enzyme activity, and calculate the 5α-reductase inhibition rate. The 5α-reductase inhibition rate (%) = inactivated enzyme activity / initial enzyme activity × 100%.

[0105] The test results are shown in Table 1. The filtrate of the rice fermentation product prepared by the method of the present invention can significantly inhibit the activity of 5α-reductase, thereby inhibiting the abnormal secretion of skin oil at the root. The present invention uses a process of enzymatic hydrolysis followed by fermentation to fully convert and utilize the active ingredients in rice. After enzymatic hydrolysis, ultrasonic treatment at a specific frequency is carried out to improve the conversion and utilization rate of the composite bacteria for the substrate. Glucoamylase and neutral protease cooperate to fully decompose the macromolecular substances in rice, and Saccharomyces cerevisiae and Lactobacillus acidophilus cooperate to carry out composite fermentation on the enzymatic hydrolysis products, producing various amino acids, polypeptides, vitamins, organic acids and minerals. The synergistic fermentation effect of Saccharomyces cerevisiae and Lactobacillus acidophilus is reflected in the two fermentation processes. In the first fermentation, metabolic conversion is initially carried out. Based on the metabolites of the first fermentation, the composite bacteria carry out a second fermentation to further produce specific active ingredients beneficial to the skin, optimize the active composition and its stability of the rice fermentation product filtrate, and enhance the oil control effect.

[0106] Table 1

[0107]

[0108]

[0109] Inhibitory rate of hyaluronidase in Test Example 2

[0110] Preparation of solutions: ① Acetate buffer solution (pH = 5.6): Take 1155 μL of glacial acetic acid in a volumetric flask, dilute it to 100 mL, mix well, and take 4.8 mL as solution a; Take 2.72 g of sodium acetate crystal, dissolve and make up the volume to 100 mL, mix well, and measure 45.2 mL as solution b; Mix solutions a and b, and make up the volume to 100 mL with deionized water and mix well; Precisely measure its pH value, and adjust the pH value to 5.6 with solution a or b; ② Hyaluronidase solution: Hyaluronidase is dissolved in the acetate buffer solution, and the final concentration of hyaluronidase is 1250 μg / mL; ③ Sodium hyaluronate solution: Sodium hyaluronate is dissolved in the acetate buffer solution, and the final concentration of sodium hyaluronate is 0.5 mg / mL; ④ Ehrlich reagent: 0.8 g of p-dimethylaminobenzaldehyde is dissolved in 15 mL of concentrated hydrochloric acid and 15 mL of absolute ethanol; ⑤ Acetylacetone solution: 3.5 mL of acetylacetone is dissolved in 50 mL of 1 M sodium carbonate solution, and the acetylacetone solution needs to be prepared freshly before use; ⑥ Test sample solution: After freeze-drying the filtrate of the rice fermentation product prepared in Examples 1-10 or Comparative Examples 1-5, add water to prepare a solution with a final concentration of 10%.

[0111] Add 0.1 mL of 0.25 mM CaCl2 solution and 0.5 mL of hyaluronidase solution into a test tube, incubate at 37 °C for 20 min; add 0.5 mL of test sample solution, incubate at 37 °C for 20 min; add 0.5 mL of sodium hyaluronate solution, incubate at 37 °C for 30 min; then let it stand at room temperature for 5 min, add 0.1 mL of 0.4 M NaOH solution and 0.5 mL of acetylacetone solution, heat and react in a boiling water bath for 15 min and then immediately ice-bath for 5 min; then add 1 mL of Ehrlich reagent and dilute it with 3 mL of absolute ethanol, let it stand for 20 min and then develop color, measure its absorbance value, calculate the hyaluronidase inhibition rate, hyaluronidase inhibition rate (%) = {[(A - B) - (C - D)] / (A - B)} × 100%, where A is the absorbance value of the control solution (using acetate buffer solution instead of test sample solution), B is the absorbance value of the blank control solution (using acetate buffer solution instead of test sample solution and enzyme solution), C is the absorbance value of the test sample solution, and D is the absorbance value of the sample blank solution (using acetate buffer solution instead of enzyme solution).

[0112] The test results are shown in Table 2. The filtrate of the rice fermentation product prepared by the method of the present invention can significantly inhibit the activity of hyaluronidase and has excellent anti-inflammatory and soothing effects; through the process of enzymatic hydrolysis first and then fermentation, the active ingredients in rice are fully transformed and utilized. After enzymatic hydrolysis, ultrasonic treatment at a specific frequency is carried out to improve the conversion and utilization rate of the complex bacteria for the substrate; glucoamylase and neutral protease synergistically decompose the macromolecular substances in rice, and Saccharomyces cerevisiae and Lactobacillus acidophilus synergistically carry out complex fermentation on the enzymatic hydrolysis products to produce various amino acids, polypeptides, vitamins, organic acids and minerals; the synergistic fermentation effect of Saccharomyces cerevisiae and Lactobacillus acidophilus is reflected in the two fermentation processes. In the first fermentation, preliminary metabolic transformation is carried out. Based on the metabolic products of the first fermentation, the complex bacteria carry out the second fermentation to further produce specific active ingredients beneficial to the skin, optimize the active composition and its stability of the rice fermentation product filtrate, and enhance the soothing effect.

[0113] Table 2

[0114]

[0115]

[0116] Test Example 3 DPPH Scavenging Rate

[0117] According to "Cosmetics - Free Radical (DPPH) Scavenging Experiment Method (T / SHRH006 - 2018)", the DPPH scavenging rates of the filtrates of the rice fermentation products prepared in Examples 1 - 10 or Comparative Examples 1 - 5 were detected respectively: A 96 - well plate was used, with 3 replicates in each group, and the system was 200 μL; after the filtrates of the rice fermentation products prepared in Examples 1 - 10 or Comparative Examples 1 - 5 were freeze - dried, a test sample solution with a final concentration of 10% was prepared by adding water. 100 μL of the test sample solution was taken, and 100 μL of 0.1 mM DPPH solution was added to the reaction system; 100 μL of distilled water + 100 μL of 0.1 mM DPPH solution was used as the control group; after the reaction system was constructed, it was shaken in the dark for 10 min, and the absorbance at 520 nm was measured using an enzyme - linked immunosorbent assay (ELISA) reader, and the DPPH scavenging rate was calculated. DPPH scavenging rate (%) = [(absorbance of the control group - absorbance of the sample group) / absorbance of the control group]×100%.

[0118] The test results are shown in Table 3. The filtrates of the rice fermentation products prepared by the method of the present invention can significantly improve the DPPH scavenging rate and have excellent antioxidant properties; through the process of enzymatic hydrolysis first and then fermentation, the active ingredients in rice are fully transformed and utilized. After enzymatic hydrolysis, ultrasonic treatment at a specific frequency is carried out to improve the conversion and utilization rate of the complex bacteria for the substrate; glucoamylase and neutral protease cooperate to fully decompose the macromolecular substances in rice, and Saccharomyces cerevisiae and Lactobacillus acidophilus cooperate to carry out complex fermentation on the enzymatic hydrolysis products, producing various amino acids, polypeptides, vitamins, organic acids and minerals; the synergistic fermentation effect of Saccharomyces cerevisiae and Lactobacillus acidophilus is reflected in the two - stage fermentation process. In the first fermentation, metabolic transformation is carried out initially. Based on the metabolic products of the first fermentation, the complex bacteria carry out the second fermentation to further produce specific active ingredients beneficial to the skin, optimize the active combination and its stability of the filtrate of the rice fermentation product, and enhance the antioxidant effect.

[0119] Table 3

[0120]

[0121]

[0122] Test Example 4 Tyrosinase Inhibition Rate

[0123] Preparation of solutions: ① Phosphate buffer: Take 250 mL of 0.2 M potassium dihydrogen phosphate solution, add 118 mL of 0.2 M sodium hydroxide solution, and make up to 1000 mL with deionized water; ② Tyrosine solution: Weigh 27.17 mg of L-tyrosine, dissolve it with phosphate buffer and make up to 100 mL in a volumetric flask to prepare a 1.5 mM L-tyrosine solution, which should be used immediately after preparation; ③ Tyrosinase solution: Weigh 18.5 mg of tyrosinase (25 KU, Solarbio), add 12.5 mL of phosphate buffer and dilute to 2 KU / mL, dispense and store frozen; dilute to 200 U / mL with phosphate buffer before use; ④ Test sample solution: After freeze-drying the filtrate of the rice fermentation product prepared in Examples 1-10 or Comparative Examples 1-5, add water to prepare a solution with a final concentration of 10%.

[0124] Under light-shielded conditions, add 50 μL of phosphate buffer, 50 μL of test sample solution, 50 μL of tyrosine solution, and 50 μL of tyrosinase solution respectively, incubate at a constant temperature of 37 °C in an enzyme-labeling instrument for 10 min, and measure the absorbance value of the sample group at 475 nm; use phosphate buffer to replace the test sample as the control group. In addition, A0 is 50 μL of tyrosine solution + 100 μL of buffer + 50 μL of tyrosinase solution, and A1 is 50 μL of tyrosine solution + 150 μL of buffer. Calculate the tyrosinase inhibition rate. Tyrosinase inhibition rate (%) = [1 - (sample group - control group) / (A0 - A1)] × 100%.

[0125] The test results are shown in Table 4. The filtrate of the rice fermentation product prepared by the method of the present invention can significantly inhibit the activity of tyrosinase and inhibit melanin production from the source; through the process of enzymatic hydrolysis first and then fermentation, the active ingredients in rice are fully transformed and utilized. After enzymatic hydrolysis, ultrasonic treatment at a specific frequency is carried out to improve the conversion and utilization rate of the complex bacteria for the substrate; glucoamylase and neutral protease cooperate to fully decompose the macromolecular substances in rice, and Saccharomyces cerevisiae and Lactobacillus acidophilus cooperate to carry out complex fermentation on the enzymatic hydrolysis products to produce various amino acids, polypeptides, vitamins, organic acids and minerals; the synergistic fermentation effect of Saccharomyces cerevisiae and Lactobacillus acidophilus is reflected in the two fermentation processes. In the first fermentation, metabolic transformation is carried out preliminarily. Based on the metabolites of the first fermentation, the complex bacteria carry out the second fermentation to further produce specific active ingredients beneficial to the skin, optimize the active composition and its stability of the filtrate of the rice fermentation product, and enhance the whitening effect.

[0126] Table 4

[0127] Test sample Tyrosinase inhibition rate (%) Example 1 89.2 Example 2 87.5 Example 3 86.3 Example 4 65.4 Example 5 63.7 Example 6 58.2 Example 7 55.9 Example 8 72.3 Example 9 78.1 Example 10 77.7 Comparative example 1 42.6 Comparative example 2 40.3 Comparative example 3 36.7 Comparative example 4 34.2 Comparative example 5 28.1

[0128] The present invention illustrates a preparation method, product and application of a soothing and oil-control rice fermentation product filtrate through the above embodiments. However, the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and disclosure scope of the present invention.

[0129] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0130] In addition, it should be noted that, in the case of no contradiction, the various specific technical features described in the above specific embodiments can be combined in any suitable manner. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A preparation method of a rice fermentation product filtrate for soothing oil control, characterized in that, The preparation method includes: (1) Mix rice and water and heat them to carry out a gelatinization reaction; (2) Adjust the pH value, add saccharifying enzyme and neutral protease to carry out an enzymatic hydrolysis reaction; (3) Inactivate the enzyme, inoculate with a compound bacterium for fermentation, sterilize after completion, and perform solid-liquid separation to obtain the filtrate of the rice fermentation product.

2. The preparation method according to claim 1, characterized in that, In step (1), the mass ratio of the rice to the water is 1:(40 - 50); Preferably, the heating temperature is 80 - 90°C and the time is 50 - 70 min.

3. The preparation method according to claim 1, characterized in that, In step (2), the pH value is 5.5 - 6.5; Preferably, the mass percentage content of the saccharifying enzyme and the neutral protease in the enzymatic hydrolysis reaction system is independently 0.01 - 1%; Preferably, the temperature of the enzymatic hydrolysis reaction is 40 - 60°C and the time is 2 - 4 h; Preferably, after the enzymatic hydrolysis reaction, there is also a step of ultrasonic treatment; Preferably, the frequency of the ultrasonic treatment is 20 - 40 kHz and the time is 10 - 30 min.

4. The preparation method according to claim 1, wherein, In step (3), the compound bacterium includes Saccharomyces cerevisiae and Lactobacillus acidophilus; Preferably, the viable count ratio of Saccharomyces cerevisiae to Lactobacillus acidophilus is (1 - 5):(1 - 5); Preferably, the fermentation includes two fermentations; Preferably, the inoculation amount of the compound bacterium in the first fermentation is 5 - 10%; Preferably, the temperature of the first fermentation is 25 - 35°C and the time is 8 - 12 h; Preferably, the inoculation amount of the compound bacterium in the second fermentation is 2 - 5%; Preferably, the temperature of the second fermentation is 28 - 30°C and the time is 24 - 48 h.

5. The preparation method according to claim 1, wherein In step (3), the temperature for enzyme inactivation is 90 - 100°C and the time is 25 - 40 min; Preferably, the temperature for sterilization is 90 - 100°C and the time is 25 - 40 min.

6. The preparation method according to claim 1, characterized in that In step (3), the method of solid-liquid separation is centrifugation; Preferably, the rotation speed of the centrifugation is 8000 - 10000 rpm and the time is 10 - 15 min; Preferably, after the solid-liquid separation, there is also a step of ultrafiltration membrane filtration.

7. The filtrate of the rice fermentation product prepared by the preparation method according to any one of claims 1 - 6.

8. Use of the filtrate of the rice fermentation product according to claim 7 in cosmetics.

9. The application according to claim 8, characterized in that, The cosmetics include any one of lotion, emulsion, cream, essence, spray, mask, sunscreen, shampoo, body wash, hair conditioner or hair mask.

10. Use of the filtrate of the rice fermentation product according to claim 7 in the preparation of a 5α-reductase inhibitor.

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