Preparation method of rice bran fermentation product filtrate as well as product and application thereof

Abstract

The invention provides a preparation method of rice bran fermentation product filtrate as well as a product and application thereof. The preparation method comprises the following steps: (1) sterilizing rice bran, mixing the sterilized rice bran with compound enzyme and water, adjusting the pH value, performing enzymolysis reaction, and performing enzyme deactivation after the enzymolysis reaction is finished to obtain rice bran zymolyte; the compound enzyme comprises alpha-amylase, beta-xylosidase and saccharifying enzyme; (2) mixing the rice bran zymolyte, an auxiliary carbon source and water to prepare a fermentation substrate; and (3) inoculating compound bacteria into the fermentation substrate for fermentation, sterilizing after fermentation, and carrying out solid-liquid separation to obtain the rice bran fermentation product filtrate. According to the method, active ingredients in the rice bran can be fully and effectively extracted and utilized, the stability and the absorption rate of the rice bran fermentation product filtrate are improved, and the prepared rice bran fermentation product filtrate contains rich effective ingredients such as ceramide, vitamins, polyphenol compounds and trace elements; the composition has excellent soothing, repairing and anti-oxidation effects.

Description

Technical Field

[0001] The invention belongs to the technical field of biological fermentation, and particularly relates to a preparation method of rice bran fermentation product filtrate, a product thereof and an application thereof. Background Art

[0002] Rice bran is a byproduct of rice processing. Composed primarily of the pericarp, seed coat, endosperm, aleurone layer, and embryo, it is rich in various nutrients and bioactive substances and is primarily used as feed. In recent years, fermented rice bran products, prepared through bio-fermentation, have been used in cosmetics to improve skin health.

[0003] CN103565724A discloses a toner containing fermented rice bran. The toner contains the following components by mass percentage: 0.01-10% fermented rice bran, 0.01-10% niacinamide, 0.1-1.0% microemulsion, 2-20% humectant, 0.05-0.2% thickener, 0.1-1.0% preservative, 0.01-1.0% pH adjuster, and the balance is water. The preparation method comprises the following steps: adding the thickener to an aqueous phase pot containing water, homogenizing and dispersing the mixture evenly, heating the mixture to 80-85°C, adding the humectant, and maintaining the mixture for 20 minutes; then stirring and cooling the mixture to 40°C, adding the pH adjuster, preservative, fermented rice bran, niacinamide, and microemulsion, and stirring evenly to obtain the toner. The toner has superior whitening, moisturizing, and anti-wrinkle effects.

[0004] CN114796079A discloses a lip beauty lotion containing rice bran essence and a preparation method thereof. The lip beauty lotion comprises the following components by weight: 0.2-5% rice bran fermentation filtrate, 0.05-1% inositol, 0.01-0.2% rice bran ceramide, 3-20% petrolatum, 0.05-0.5% emulsifier, 0.05-0.5% antioxidant, 0.05-0.5% humectant, 1-8% emollient, 0.01-0.5% preservative, and the balance hydrogenated isobutylene. The rice bran fermentation filtrate is obtained by mixing rice bran with desalinated deep ocean water, enzymatically hydrolyzing with amylase and protease, and then fermenting with a culture of Saccharomyces cerevisiae seed, centrifuging, and filtering the supernatant. During preparation, rice bran ceramide, inositol, and rice bran fermentation filtrate are uniformly mixed; hydrogenated isobutylene and petrolatum are added and mixed uniformly; a moisturizer, emollient, antioxidant, emulsifier, and preservative are added to obtain a lip beauty serum. The beauty serum has the advantages of effectively moisturizing the lip skin, promoting dermal regeneration and repair, improving lip skin elasticity, and reducing lip lines.

[0005] However, the water solubility and absorbability of existing rice bran fermentation products need to be improved, and the rice bran fermentation technology is not perfect. Based on this, it is urgent to improve the rice bran fermentation conditions so that the active ingredients in the rice bran can be effectively extracted and utilized. Summary of the Invention

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a method for preparing a rice bran fermentation product filtrate, as well as its product and application. The method of the present invention can significantly improve the stability, water solubility and absorbability of the rice bran fermentation product filtrate, and increase the content of active ingredients. The prepared rice bran fermentation product filtrate has excellent repairing, soothing and antioxidant effects.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a method for preparing a rice bran fermentation product filtrate, the preparation method comprising:

[0009] (1) sterilizing rice bran, mixing it with a composite enzyme and water, adjusting the pH value, performing an enzymatic hydrolysis reaction, and then inactivating the enzyme to obtain a rice bran hydrolyzate; the composite enzyme comprising α-amylase, β-xylosidase, and saccharifying enzyme;

[0010] (2) mixing the rice bran hydrolysate, the auxiliary carbon source and water to prepare a fermentation matrix;

[0011] (3) inoculating composite bacteria into the fermentation matrix for fermentation, sterilizing after the fermentation, and performing solid-liquid separation to obtain the rice bran fermentation product filtrate.

[0012] The method of the present invention can fully and effectively extract and utilize the active ingredients in rice bran, improving the stability and absorption rate of the rice bran fermentation product filtrate. The prepared rice bran fermentation product filtrate is rich in active ingredients such as ceramides, vitamins, polyphenols, and trace elements, and has excellent soothing, repairing, and antioxidant effects. The synergistic action of α-amylase, β-xylosidase, and saccharifying enzyme can effectively carry out the enzymatic hydrolysis reaction of rice bran, providing a sufficient substrate source for the fermentation of the composite bacteria.

[0013] Preferably, in step (1), the mass ratio of rice bran to water is 1:(10-15).

[0014] The specific point values ​​in (10-15) can be 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5 or 15, etc.

[0015] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0016] Preferably, the mass percentage of the complex enzyme in the enzymatic reaction system is 0.1-1%, for example, it can be 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%, etc.

[0017] Preferably, the pH value is 5-7, for example, it can be 5, 5.2, 5.4, 5.6, 5.8, 6, 6.2, 6.4, 6.6, 6.8 or 7.

[0018] 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; the time is 3-6h, for example, it can be 3h, 3.2h, 3.5h, 3.8h, 4h, 4.2h, 4.5h, 4.8h, 5h, 5.2h, 5.5h, 5.8h or 6h.

[0019] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0020] Preferably, the mass ratio of the α-amylase, β-xylosidase and saccharifying enzyme is (1-3):(1-3):(2-5).

[0021] The specific point values ​​in the first one (1-3) can be 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8 or 3, etc.

[0022] The specific point values ​​in the second one (1-3) can be 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8 or 3, etc.

[0023] The specific point values ​​in (2-5) 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.

[0024] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0025] Preferably, in step (1), the sterilization temperature is 120-140°C, for example, it can be 120°C, 122°C, 124°C, 126°C, 128°C, 130°C, 132°C, 134°C, 136°C, 138°C or 140°C; the time is 20-40min, for example, it can be 20min, 22min, 24min, 26min, 28min, 30min, 32min, 34min, 36min, 38min or 40min, etc.

[0026] Preferably, the enzyme inactivation temperature is 120-140°C, for example, it can be 120°C, 122°C, 124°C, 126°C, 128°C, 130°C, 132°C, 134°C, 136°C, 138°C or 140°C; the time is 20-40 min, for example, it can be 20 min, 22 min, 24 min, 26 min, 28 min, 30 min, 32 min, 34 min, 36 min, 38 min or 40 min.

[0027] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0028] Preferably, in step (2), the mass ratio of the rice bran hydrolysate, the auxiliary carbon source and water is (1-2):(0.1-1):(2-4).

[0029] The specific point values ​​in (1-2) can be 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2, etc.

[0030] The specific point values ​​in (0.1-1) can be 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1, etc.

[0031] The specific point values ​​in (2-4) can be 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.4, 3.6, 3.8 or 4, etc.

[0032] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0033] Preferably, the auxiliary carbon source includes any one of glucose, lactose, mannose, fructose, xylose or arabinose, or a combination of at least two of them.

[0034] Preferably, the auxiliary carbon source comprises glucose, lactose and mannose.

[0035] Auxiliary carbon source is an important component of fermentation carbon source. Glucose, lactose and mannose have a synergistic effect and can better assist rice bran hydrolysate in providing sufficient substrate source for composite bacteria fermentation.

[0036] Preferably, the mass ratio of glucose, lactose and mannose is (1-5):(2-4):(2-4).

[0037] The specific point values ​​in (1-5) can be 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5, etc.

[0038] The specific point values ​​in the first one (2-4) can be 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.4, 3.6, 3.8 or 4, etc.

[0039] The specific point values ​​in the second one (2-4) can be 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.4, 3.6, 3.8 or 4, etc.

[0040] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0041] Preferably, in step (3), the composite bacteria include Saccharomyces cerevisiae, Aspergillus oryzae and Lactobacillus plantarum.

[0042] Saccharomyces cerevisiae, Aspergillus oryzae and Lactobacillus plantarum work together to ferment rice bran, which can fully convert the fermentation matrix into active ingredients that are easy to absorb, enhancing the soothing, repairing and antioxidant effects of the rice bran fermentation product filtrate.

[0043] Preferably, the ratio of the viable counts of Saccharomyces cerevisiae, Aspergillus oryzae and Lactobacillus plantarum is (3-6):(1-5):(2-4).

[0044] The specific point values ​​in (3-6) can be 3, 3.2, 3.5, 3.8, 4, 4.2, 4.5, 4.8, 5, 5.2, 5.5, 5.8 or 6, etc.

[0045] The specific point values ​​in (1-5) can be 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5, etc.

[0046] The specific point values ​​in (2-4) can be 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.4, 3.6, 3.8 or 4, etc.

[0047] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0048] Preferably, in step (3), the fermentation comprises performing two fermentations.

[0049] Preferably, the inoculation amount of the composite bacteria in the first fermentation is 6-9%, for example, it can be 6%, 6.2%, 6.5%, 6.8%, 7%, 7.2%, 7.5%, 7.8%, 8%, 8.2%, 8.5%, 8.8% or 9%.

[0050] Preferably, the pH of the first fermentation is 5-6, for example, it can be 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 or 6; the temperature is 23-28°C, for example, it can be 23°C, 23.5°C, 24°C, 24.5°C, 25°C, 25.5°C, 26°C, 26.5°C, 27°C, 27.5°C or 28°C; the time is 10-16h, for example, it can be 10h, 10.5h, 11h, 11.5h, 12h, 12.5h, 13h, 13.5h, 14h, 14.5h, 15h, 15.5h or 16h.

[0051] Preferably, the inoculation amount of the composite bacteria in the second fermentation is 10-15%, for example, it can be 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5% or 15%.

[0052] Preferably, the pH of the second fermentation is 6.5-7.5, for example, it can be 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4 or 7.5; the temperature is 30-35°C, for example, it can be 30°C, 30.5°C, 31°C, 31.5°C, 32°C, 32.5°C, 33°C, 33.5°C, 34°C, 34.5°C or 35°C; the time is 20-30h, for example, it can be 20h, 21h, 22h, 23h, 24h, 25h, 26h, 27h, 28h, 29h or 30h, etc.

[0053] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0054] The complex bacteria undergo a two-stage gradient fermentation process, first converting the fermentation matrix, consisting of rice bran hydrolysate and a supplemental carbon source, into small-molecule active ingredients. A second fermentation reaction, based on the initial fermentation product, then generates more readily absorbed active ingredients. This gradient fermentation process effectively extracts and utilizes the active ingredients in the rice bran, enhancing the soothing, repairing, and antioxidant properties of the rice bran fermentation filtrate.

[0055] Preferably, in step (3), the solid-liquid separation is performed by centrifugation.

[0056] Preferably, the centrifugal speed is 9000-11000 rpm, for example, it can be 9000 rpm, 9200 rpm, 9400 rpm, 9600 rpm, 9800 rpm, 10000 rpm, 10200 rpm, 10400 rpm, 10600 rpm, 10800 rpm or 11000 rpm, etc.; the time is 8-20 min, for example, it can be 8 min, 9 min, 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, 16 min, 17 min, 18 min, 19 min or 20 min, etc.

[0057] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0058] Preferably, the solid-liquid separation further includes a step of filtering through an ultrafiltration membrane.

[0059] Preferably, in step (3), the sterilization temperature is 120-140°C, for example, it can be 120°C, 122°C, 124°C, 126°C, 128°C, 130°C, 132°C, 134°C, 136°C, 138°C or 140°C; the time is 20-40min, for example, it can be 20min, 22min, 24min, 26min, 28min, 30min, 32min, 34min, 36min, 38min or 40min.

[0060] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0061] In a second aspect, the present invention provides a rice bran fermentation product filtrate prepared by the preparation method according to the first aspect.

[0062] In a third aspect, the present invention provides use of the rice bran fermentation product filtrate as described in the second aspect in cosmetics.

[0063] Preferably, the cosmetics include any one of lotion, emulsion, cream, essence, spray, mask, sunscreen, shampoo, shower gel, conditioner or hair mask.

[0064] The rice bran fermentation product filtrate provided by the present invention can be used not only in skin care products but also in shampoo and conditioner products, especially in shampoo and conditioner products for hair care, and has the effects of mildly soothing, nourishing hair follicles, and strengthening hair and preventing hair loss.

[0065] Compared with the prior art, the present invention has the following beneficial effects:

[0066] The method of the present invention can fully and effectively extract and utilize the active ingredients in rice bran, improving the stability and absorption rate of the rice bran fermentation product filtrate. The prepared rice bran fermentation product filtrate is rich in active ingredients such as ceramides, vitamins, polyphenols, and trace elements, and has excellent soothing, repairing, and antioxidant effects. The synergistic action of α-amylase, β-xylosidase, and saccharifying enzyme can effectively carry out the enzymatic hydrolysis reaction of rice bran, providing a sufficient substrate source for the fermentation of the composite bacteria. DETAILED DESCRIPTION

[0067] In order to further illustrate the technical means and effects adopted by the present invention, the technical solutions of the present invention are further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0068] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0069] The sources of materials used in the following specific embodiments are as follows:

[0070] Raw material name Purchase manufacturer model α-amylase Dalian Meilun Biotechnology Co., Ltd. MB3100 β-Xylosidase Shanghai Jingke Chemical Technology Co., Ltd. E-BXSEBP Glucoamylase Dalian Meilun Biotechnology Co., Ltd. MB2585 Saccharomyces cerevisiae China Industrial Microbiological Culture Collection Management Center CICC31105 Aspergillus oryzae China Industrial Microbiological Culture Collection Management Center CICC2013 Lactobacillus plantarum China Industrial Microbiological Culture Collection Management Center CICC22201

[0071] Example 1

[0072] This embodiment provides a method for preparing a rice bran fermentation product filtrate, the preparation method comprising:

[0073] (1) Wash the rice bran with water to remove impurities, dry it, grind it through a 100-mesh sieve, and keep it at 130°C for 30 minutes to remove bacteria; mix the sterilized rice bran with water at a mass ratio of 1:13, add 0.2% α-amylase, 0.2% β-xylosidase, and 0.3% saccharifying enzyme, adjust the pH to 6, and react at 50°C for 4 hours; after the enzymatic hydrolysis reaction is completed, keep it at 130°C for 30 minutes to inactivate the enzyme, and obtain a rice bran hydrolysate;

[0074] (2) mixing rice bran hydrolysate, an auxiliary carbon source, and water in a mass ratio of 1.5:0.5:3 to prepare a fermentation matrix; the auxiliary carbon source is glucose, lactose, and mannose in a mass ratio of 3:3:4;

[0075] (3) The fermentation matrix was inoculated with 4% activated bacterial solution of Saccharomyces cerevisiae, 2% activated bacterial solution of Aspergillus oryzae and 3% activated bacterial solution of Lactobacillus plantarum (the concentration of the bacterial solution was 1×10 10 CFU / mL), the first fermentation was carried out at pH = 5.5 and 25 ° C for 13 h; then 5% activated bacterial solution of Saccharomyces cerevisiae, 4% activated bacterial solution of Aspergillus oryzae and 3% activated bacterial solution of Lactobacillus plantarum were inoculated (the concentration of bacterial solution was 1×10 10CFU / mL), and a second fermentation was carried out at pH = 7 and 34° C. for 25 h; after the fermentation reaction, the mixture was kept at 130° C. for 30 min for sterilization; and centrifuged at 10,000 rpm for 15 min, the liquid was collected, and filtered through an ultrafiltration membrane to obtain the rice bran fermentation product filtrate.

[0076] Example 2

[0077] This embodiment provides a method for preparing a rice bran fermentation product filtrate, the preparation method comprising:

[0078] (1) Wash the rice bran with water to remove impurities, dry it, grind it through a 100-mesh sieve, and keep it at 120°C for 40 minutes to remove bacteria; mix the sterilized rice bran with water at a mass ratio of 1:15, add 0.1% α-amylase, 0.3% β-xylosidase, and 0.2% saccharifying enzyme, adjust the pH to 5, and react at 60°C for 3 hours; after the enzymatic hydrolysis reaction is completed, keep it at 120°C for 40 minutes to inactivate the enzyme, and obtain a rice bran hydrolyzate;

[0079] (2) mixing rice bran hydrolysate, an auxiliary carbon source, and water in a mass ratio of 1:0.1:2 to prepare a fermentation matrix; the auxiliary carbon source is glucose, lactose, and mannose in a mass ratio of 1:2:2;

[0080] (3) The fermentation matrix was inoculated with 3% activated bacterial solution of Saccharomyces cerevisiae, 2% activated bacterial solution of Aspergillus oryzae and 4% activated bacterial solution of Lactobacillus plantarum (the concentration of the bacterial solution was 1×10 10 CFU / mL), the first fermentation was carried out at pH = 5 and 23 ° C for 16 h; then 6% activated bacterial solution of Saccharomyces cerevisiae, 3% activated bacterial solution of Aspergillus oryzae and 2% activated bacterial solution of Lactobacillus plantarum were inoculated (the concentration of bacterial solution was 1 × 10 10 CFU / mL), and a second fermentation was carried out at pH = 6.5 and 30° C. for 30 h; after the fermentation reaction, the mixture was kept at 120° C. for 40 min for sterilization; and centrifuged at 9000 rpm for 20 min, the liquid was collected, and filtered through an ultrafiltration membrane to obtain the rice bran fermentation product filtrate.

[0081] Example 3

[0082] This embodiment provides a method for preparing a rice bran fermentation product filtrate, the preparation method comprising:

[0083] (1) Wash the rice bran with water to remove impurities, dry it, grind it through a 100-mesh sieve, and keep it at 140°C for 20 minutes to remove bacteria; mix the sterilized rice bran with water at a mass ratio of 1:10, add 0.3% α-amylase, 0.2% β-xylosidase, and 0.5% saccharifying enzyme, adjust the pH to 7, and react at 40°C for 6 hours; after the enzymatic hydrolysis reaction is completed, keep it at 140°C for 20 minutes to inactivate the enzyme, and obtain a rice bran hydrolysate;

[0084] (2) mixing rice bran hydrolysate, an auxiliary carbon source, and water in a mass ratio of 2:1:4 to prepare a fermentation matrix; the auxiliary carbon source is glucose, lactose, and mannose in a mass ratio of 5:4:3;

[0085] (3) The fermentation matrix was inoculated with 5% activated bacterial solution of Saccharomyces cerevisiae, 1% activated bacterial solution of Aspergillus oryzae, and 2% activated bacterial solution of Lactobacillus plantarum (the concentration of the bacterial solution was 1×10 10 CFU / mL), the first fermentation was carried out at pH = 6 and 28 ° C for 10 h; then 3% activated bacterial solution of Saccharomyces cerevisiae, 5% activated bacterial solution of Aspergillus oryzae and 2% activated bacterial solution of Lactobacillus plantarum were inoculated (the concentration of bacterial solution was 1×10 10 CFU / mL), and a second fermentation was carried out at pH = 7.5 and 35° C. for 20 h; after the fermentation reaction, the mixture was kept at 140° C. for 20 min for sterilization; and centrifuged at 11000 rpm for 8 min, the liquid was collected, and filtered through an ultrafiltration membrane to obtain the rice bran fermentation product filtrate.

[0086] Example 4

[0087] This embodiment provides a method for preparing a rice bran fermentation product filtrate, the preparation method comprising:

[0088] (1) Wash the rice bran with water to remove impurities, dry it, grind it through a 100-mesh sieve, and keep it at 130°C for 30 minutes to remove bacteria; mix the sterilized rice bran with water at a mass ratio of 1:13, add 0.2% α-amylase, 0.2% β-xylosidase, and 0.3% saccharifying enzyme, adjust the pH to 6, and react at 50°C for 4 hours; after the enzymatic hydrolysis reaction is completed, keep it at 130°C for 30 minutes to inactivate the enzyme, and obtain a rice bran hydrolysate;

[0089] (2) mixing rice bran hydrolysate, an auxiliary carbon source, and water in a mass ratio of 1.5:0.5:3 to prepare a fermentation matrix; the auxiliary carbon source is glucose, lactose, and mannose in a mass ratio of 3:3:4;

[0090] (3) The fermentation matrix was inoculated with 5% activated bacterial solution of Saccharomyces cerevisiae, 4% activated bacterial solution of Aspergillus oryzae and 3% activated bacterial solution of Lactobacillus plantarum (the concentration of bacterial solution was 1×10 10 CFU / mL), the first fermentation was carried out at pH = 7 and 34 ° C for 25 h; then 4% activated bacterial solution of Saccharomyces cerevisiae, 2% activated bacterial solution of Aspergillus oryzae and 3% activated bacterial solution of Lactobacillus plantarum were inoculated (the concentration of bacterial solution was 1 × 10 10 CFU / mL), and a second fermentation was carried out at pH = 5.5 and 25° C. for 13 h; after the fermentation reaction, the mixture was kept at 130° C. for 30 min for sterilization; and centrifuged at 10,000 rpm for 15 min, the liquid was collected, and filtered through an ultrafiltration membrane to obtain the rice bran fermentation product filtrate.

[0091] Example 5

[0092] This embodiment provides a method for preparing a rice bran fermentation product filtrate, the preparation method comprising:

[0093] (1) Wash the rice bran with water to remove impurities, dry it, grind it through a 100-mesh sieve, and keep it at 130°C for 30 minutes to remove bacteria; mix the sterilized rice bran with water at a mass ratio of 1:13, add 0.2% α-amylase, 0.2% β-xylosidase, and 0.3% saccharifying enzyme, adjust the pH to 6, and react at 50°C for 4 hours; after the enzymatic hydrolysis reaction is completed, keep it at 130°C for 30 minutes to inactivate the enzyme, and obtain a rice bran hydrolysate;

[0094] (2) mixing rice bran hydrolysate, an auxiliary carbon source, and water in a mass ratio of 1.5:0.5:3 to prepare a fermentation matrix; the auxiliary carbon source is glucose, lactose, and mannose in a mass ratio of 3:3:4;

[0095] (3) The fermentation matrix was inoculated with 9% activated bacterial solution of Saccharomyces cerevisiae, 6% activated bacterial solution of Aspergillus oryzae and 6% activated bacterial solution of Lactobacillus plantarum (the concentration of the bacterial solution was 1×10 10 CFU / mL), only one fermentation was carried out, pH = 5.5, and fermentation was carried out at 25° C. for 38 hours; after the fermentation reaction, the mixture was kept at 130° C. for 30 minutes for sterilization; centrifugation was carried out at 10,000 rpm for 15 minutes, and the liquid was collected and filtered through an ultrafiltration membrane to obtain the rice bran fermentation product filtrate.

[0096] Example 6

[0097] This embodiment provides a method for preparing a rice bran fermentation product filtrate, the preparation method comprising:

[0098] (1) Wash the rice bran with water to remove impurities, dry it, grind it through a 100-mesh sieve, and keep it at 130°C for 30 minutes to remove bacteria; mix the sterilized rice bran with water at a mass ratio of 1:13, add 0.2% α-amylase, 0.2% β-xylosidase, and 0.3% saccharifying enzyme, adjust the pH to 6, and react at 50°C for 4 hours; after the enzymatic hydrolysis reaction is completed, keep it at 130°C for 30 minutes to inactivate the enzyme, and obtain a rice bran hydrolysate;

[0099] (2) mixing rice bran hydrolysate, an auxiliary carbon source, and water in a mass ratio of 1.5:0.5:3 to prepare a fermentation matrix; the auxiliary carbon source is glucose, lactose, and mannose in a mass ratio of 3:3:4;

[0100] (3) The fermentation matrix was inoculated with 9% activated bacterial solution of Saccharomyces cerevisiae, 6% activated bacterial solution of Aspergillus oryzae and 6% activated bacterial solution of Lactobacillus plantarum (the concentration of the bacterial solution was 1×10 10CFU / mL), only one fermentation was carried out, pH = 7, and fermentation was carried out at 34° C. for 38 hours; after the fermentation reaction, the mixture was kept at 130° C. for 30 minutes for sterilization; centrifugation was carried out at 10,000 rpm for 15 minutes, and the liquid was collected and filtered through an ultrafiltration membrane to obtain the rice bran fermentation product filtrate.

[0101] Example 7

[0102] This embodiment provides a method for preparing a rice bran fermentation product filtrate, which differs from Example 1 only in that: in step (3), no activated bacterial solution of Saccharomyces cerevisiae is added, and the reduced amount is proportionally distributed to the activated bacterial solution of Aspergillus oryzae and the activated bacterial solution of Lactobacillus plantarum, and the remaining steps remain unchanged.

[0103] Example 8

[0104] This embodiment provides a method for preparing a rice bran fermentation product filtrate, which differs from Example 1 only in that: in step (3), no activated Aspergillus oryzae bacterial solution is added, and the reduced amount is proportionally distributed to the activated Saccharomyces cerevisiae bacterial solution and the activated Lactobacillus plantarum bacterial solution, and the remaining steps remain unchanged.

[0105] Example 9

[0106] This embodiment provides a method for preparing a rice bran fermentation product filtrate, which differs from Example 1 only in that: in step (3), no activated Lactobacillus plantarum bacterial solution is added, and the reduced amount is proportionally distributed to the activated Saccharomyces cerevisiae bacterial solution and the activated Aspergillus oryzae bacterial solution, and the remaining steps remain unchanged.

[0107] Example 10

[0108] This embodiment provides a method for preparing a filtrate of a rice bran fermentation product, which differs from Example 1 only in that, in step (2), glucose is not added, and the reduced amount thereof is proportionally distributed to lactose and mannose, and the remaining steps remain unchanged.

[0109] Example 11

[0110] This embodiment provides a method for preparing a filtrate of a rice bran fermentation product, which differs from Example 1 only in that, in step (2), lactose is not added, and the reduced amount thereof is proportionally distributed to glucose and mannose, and the remaining steps remain unchanged.

[0111] Example 12

[0112] This embodiment provides a method for preparing a filtrate of a rice bran fermentation product, which differs from Example 1 only in that, in step (2), no mannose is added, and the reduced amount thereof is proportionally distributed to glucose and lactose, and the remaining steps remain unchanged.

[0113] Example 13

[0114] This embodiment provides a method for preparing a filtrate of a rice bran fermentation product, which differs from Example 1 only in that: in step (2), "the auxiliary carbon source is glucose, lactose and mannose in a mass ratio of 3:3:4" is replaced by "the auxiliary carbon source is glucose, galactose and maltose in a mass ratio of 3:3:4", and the remaining steps remain unchanged.

[0115] Comparative Example 1

[0116] This comparative example provides a method for preparing a rice bran fermentation product filtrate, which differs from Example 1 only in that: in step (1), no α-amylase is added, and its reduced amount is proportionally distributed to β-xylosidase and saccharifying enzyme, and the remaining steps remain unchanged.

[0117] Comparative Example 2

[0118] This comparative example provides a method for preparing a rice bran fermentation product filtrate, which differs from Example 1 only in that: in step (1), β-xylosidase is not added, and its reduced amount is proportionally distributed to α-amylase and saccharifying enzyme, and the remaining steps remain unchanged.

[0119] Comparative Example 3

[0120] This comparative example provides a method for preparing a rice bran fermentation product filtrate, which differs from Example 1 only in that: in step (1), no saccharifying enzyme is added, and its reduced amount is proportionally distributed to α-amylase and β-xylosidase, and the remaining steps remain unchanged.

[0121] Test Example 1 Antioxidant Performance

[0122] The rice bran fermentation product filtrates obtained in Examples 1-13 and Comparative Examples 1-3 were freeze-dried and then water was added to prepare test sample solutions with a final concentration of 5%. 2 mL of the test sample solution and 2 mL of the DPPH solution were respectively taken into a stoppered test tube, mixed, reacted in the dark for 30 minutes, and the absorbance value A1 at a wavelength of 517 nm was measured; 2 mL of the test sample solution and 2 mL of methanol were respectively taken into a stoppered test tube, mixed, reacted in the dark for 30 minutes, and the absorbance value A2 at a wavelength of 517 nm was measured; 2 mL of the DPPH solution and 2 mL of methanol were respectively taken into a stoppered test tube, mixed, reacted in the dark for 30 minutes, and the absorbance value A0 at a wavelength of 517 nm was measured; each sample

[0123]

[0124] The test results are shown in Table 1: The rice bran fermentation product filtrate prepared by the method of the present invention significantly improved the DPPH free radical scavenging rate and exhibited excellent antioxidant properties. α-amylase, β-xylosidase, and saccharifying enzymes synergistically perform enzymatic hydrolysis reactions, while glucose, lactose, and mannose serve as auxiliary carbon sources, promoting the fermentation of the composite bacteria to produce more active ingredients that are easily absorbed. Saccharomyces cerevisiae, Aspergillus oryzae, and Lactobacillus plantarum synergistically ferment rice bran using a two-stage gradient fermentation process, fully transforming the fermentation substrate and enhancing the antioxidant efficacy of the rice bran fermentation product filtrate.

[0125] Table 1

[0126]

[0127]

[0128] Test Example 2 Soothing Repair

[0129] The rice bran fermentation product filtrates obtained in Examples 1-13 and Comparative Examples 1-3 were freeze-dried and then added to water to prepare test sample solutions with a final concentration of 5%. The anti-allergic activity and soothing and repairing efficacy of the test sample solutions were evaluated by measuring the change in absorbance and calculating the hyaluronidase inhibition rate, as described in T / JSRH 003-2023, Cosmetic Ingredient Anti-Allergic Activity Test - Hyaluronidase Inhibition Method.

[0130] The test results are shown in Table 2: The rice bran fermentation product filtrate prepared by the method of the present invention significantly increased the hyaluronidase inhibition rate and exhibited excellent soothing and repairing properties. α-amylase, β-xylosidase, and saccharifying enzymes synergize in the enzymatic hydrolysis reaction, while glucose, lactose, and mannose serve as auxiliary carbon sources, promoting the fermentation of the complex bacteria to produce more active ingredients that are easily absorbed. Saccharomyces cerevisiae, Aspergillus oryzae, and Lactobacillus plantarum collaborate in the rice bran fermentation process based on a two-stage gradient fermentation process, fully transforming the fermentation substrate and enhancing the soothing and repairing effects of the rice bran fermentation product filtrate.

[0131] Table 2

[0132]

[0133]

[0134] The present invention uses the above-described embodiments to illustrate the method for preparing a rice bran fermentation product filtrate, its product, and its applications. However, the present invention is not limited to the above-described embodiments, and implementation of the present invention is not necessarily dependent on the above-described embodiments. Those skilled in the art will appreciate that any improvements to the present invention, equivalent substitutions for various raw materials in the product of the present invention, addition of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

[0135] The preferred embodiments of the present invention are 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 solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0136] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A method for preparing a rice bran fermentation product filtrate, characterized in that: The preparation method comprises: (1) sterilizing rice bran, mixing it with a composite enzyme and water, adjusting the pH value, performing an enzymatic hydrolysis reaction, and then inactivating the enzyme to obtain a rice bran hydrolyzate; the composite enzyme comprising α-amylase, β-xylosidase, and saccharifying enzyme; (2) mixing the rice bran hydrolysate, the auxiliary carbon source and water to prepare a fermentation matrix; (3) inoculating composite bacteria into the fermentation matrix for fermentation, sterilizing after fermentation, and performing solid-liquid separation to obtain the rice bran fermentation product filtrate.

2. The preparation method according to claim 1, characterized in that In step (1), the mass ratio of rice bran to water is 1:(10-15); Preferably, the mass percentage of the complex enzyme in the enzymatic reaction system is 0.1-1%; Preferably, the pH value is 5-7; Preferably, the temperature of the enzymatic hydrolysis reaction is 40-60°C and the time is 3-6h; Preferably, the mass ratio of the α-amylase, β-xylosidase and saccharifying enzyme is (1-3):(1-3):(2-5).

3. The preparation method according to claim 1 or 2, characterized in that In step (1), the sterilization temperature is 120-140°C and the time is 20-40 minutes; Preferably, the enzyme inactivation temperature is 120-140° C. and the time is 20-40 min.

4. The preparation method according to any one of claims 1 to 3, characterized in that In step (2), the mass ratio of the rice bran hydrolysate, the auxiliary carbon source and water is (1-2):(0.1-1):(2-4); Preferably, the auxiliary carbon source comprises any one or a combination of at least two of glucose, lactose, mannose, fructose, xylose or arabinose; Preferably, the auxiliary carbon source comprises glucose, lactose and mannose; Preferably, the mass ratio of glucose, lactose and mannose is (1-5):(2-4):(2-4).

5. The preparation method according to any one of claims 1 to 4, characterized in that In step (3), the composite bacteria include Saccharomyces cerevisiae, Aspergillus oryzae and Lactobacillus plantarum; Preferably, the ratio of the viable counts of Saccharomyces cerevisiae, Aspergillus oryzae and Lactobacillus plantarum is (3-6):(1-5):(2-4).

6. The preparation method according to any one of claims 1 to 5, characterized in that In step (3), the fermentation includes performing two fermentations; Preferably, the inoculation amount of the composite bacteria in the first fermentation is 6-9%; Preferably, the pH of the first fermentation is 5-6, the temperature is 23-28°C, and the time is 10-16 hours; Preferably, the inoculation amount of the composite bacteria in the second fermentation is 10-15%; Preferably, the pH of the second fermentation is 6.5-7.5, the temperature is 30-35° C., and the time is 20-30 h.

7. The preparation method according to any one of claims 1 to 6, characterized in that In step (3), the solid-liquid separation method is centrifugation; Preferably, the centrifugal speed is 9000-11000 rpm and the time is 8-20 min; Preferably, after the solid-liquid separation, the step of filtering through an ultrafiltration membrane is further included; Preferably, in step (3), the sterilization temperature is 120-140° C. and the time is 20-40 min.

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

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

10. The use according to claim 9, characterized in that The cosmetics include any one of lotion, emulsion, cream, essence, spray, mask, sunscreen, shampoo, shower gel, conditioner or hair mask.

Images