Multi-dimensional fermented essence / powder as well as preparation method and application thereof

Abstract

The invention discloses a multi-vitamin fermentation essence which is prepared from the following components in percentage by mass: 0.1%-30% of polyglutamic acid, 0.01%-15.6% of fructan LEVAN, 0.2 ppm to 94 ppm of vitamin K2 and 0.02 ppm to 32 ppm of folic acid, and the molecular weight of the polyglutamic acid is 400D to 3,000,000 D. The invention further discloses a preparation method of the multi-vitamin fermentation essence. The invention further provides a preparation method of the multi-dimensional fermentation essence. The preparation method comprises the following steps: preparing bacillus subtilis fermentation liquor; adjusting the pH value of the bacillus subtilis fermentation liquor to 3.9 to 4.5 by using acid; adding diatomite, standing and settling, and removing the precipitate; the obtained supernate is filtered through a membrane, protein and salt are removed, washing and decolorizing are conducted, and the multi-dimensional fermentation essence is obtained. The invention further discloses a cosmetic which contains the multi-dimensional fermented essence or the multi-dimensional fermented essence powder.

Description

[0001] This application is a divisional application of an application with an application date of January 11, 2019, application number 201910024963.8, and invention name “A multidimensional fermentation essence liquid / powder and its preparation method and application”. Technical Field

[0002] The present application relates to the field of cosmetics, and in particular to a preparation method of a multi-dimensional fermentation essence liquid / powder and its application. Background Art

[0003] The functional raw materials of cosmetics mainly include cosmetic Chinese medicine, biological products, and natural functional raw materials. Cosmetic Chinese medicine has both nutritional and medicinal effects, and has certain advantages over synthetic raw materials. However, the ingredients of cosmetic Chinese medicine are relatively complex. There are several or even dozens of ingredients in one Chinese medicine, and the ingredients for improving skin are relatively less. The use of chemical methods to extract the effective substances may leave other useless organic solvents. And some Chinese medicines are toxic, such as blister beetles. Some Chinese medicines contain carcinogenic volatile oils, such as asarum and cloves. However, the research on the ingredients in Chinese medicine is incomplete at this stage, and we cannot be sure that the added pure natural Chinese medicine is completely harmless to our skin. Most of the cosmetic raw materials of biological products come from animals, and the viruses that animals may carry bring a hint of uncertainty to the safety of cosmetic raw materials. And the cosmetic raw materials of biological products on the market are relatively expensive due to the source of materials and extraction methods. Although natural functional raw materials are advertised as natural, most of the raw materials are extracted using chemical methods using some organic solvents during the extraction process, which may lead to organic solvent residues. Summary of the invention

[0004] The following is an overview of the subject matter described in detail herein. This overview is not intended to limit the scope of protection of this application.

[0005] The research and application of Bacillus subtilis has a history of more than 100 years. With the in-depth development of the research on Bacillus subtilis, it has been found that it has wide application value in industry, agriculture, medicine and health, food health care, aquaculture, etc. Bacillus subtilis enzyme is one of the most widely used bacteria in industrial enzyme production today. The enzyme produced by its fermentation plays a very important role in food, feed, washing, textile, leather, papermaking and medicine. The source of cosmetic raw materials obtained by fermentation of Bacillus subtilis is clear, not through chemical synthesis, and the method of obtaining raw materials is to process them through advanced membrane technology without adding organic solvents, which is truly harmless and non-toxic to the skin.

[0006] The present application provides a multi-dimensional fermentation essence liquid / powder, which has significantly increased whitening, moisturizing, anti-wrinkle and freckle removal effects.

[0007] Specifically, the present application provides a multi-dimensional fermentation essence, which comprises, by mass fraction, 0.1%-30% polyglutamic acid, 0.01%-15.6% fructan LEVAN, 0.2 ppm-94 ppm vitamin K2, and 0.02 ppm-32 ppm folic acid, where ppm represents the unit of concentration, one millionth of a gram, wherein the molecular weight of the polyglutamic acid is 400D-3,000,000D.

[0008] Levan is a type of polysaccharide that contains a large number of β-(2,6) glycosidic bonds and a small number of β-(2,1) fructosidic bonds in its molecules.

[0009] In the present application, the multi-dimensional fermentation essence contains, by mass fraction, 0.1%-30% polyglutamic acid, 4%-10% LEVE polysaccharide, 40 ppm-94 ppm vitamin K2, and 0.02 ppm-32 ppm folic acid, where ppm represents the unit of concentration, one millionth of a gram, and the molecular weight of the polyglutamic acid is 2,000-1,000,000D.

[0010] The present application also provides a method for preparing the multidimensional fermentation essence, which comprises: (1) Preparation of Bacillus subtilis fermentation broth: a Bacillus subtilis suspension having a concentration of 5% to 20% is placed in a seed tank containing LB solution for expansion. After the seed solution concentration reaches a bacterial activity of OD600 between 1.2 and 2.2, the suspension is placed in a fermentation tank containing fermentation broth and fermented at 35° C. to 50° C. for 10 to 48 hours to obtain the Bacillus subtilis fermentation broth. (2) adjusting the pH of the Bacillus subtilis fermentation broth to 3.9-4.5 with acid; (3) Add diatomaceous earth, let it settle, and discard the precipitate; (4) The supernatant obtained in step (3) is filtered through a membrane to remove protein and salt, and then washed and decolorized with water to obtain the multi-dimensional fermentation essence.

[0011] In the present application, in step (1), the Bacillus subtilis is isolated from natto and is a wild-type strain of Bacillus subtilis natto ST188, CGMCC No. 8400.

[0012] In the present application, in step (1), the LB liquid is Luria-Bertani liquid culture medium: calculated by mass volume ratio g / 100 ml water, it contains 0.1% glucose, 1% yeast powder, 0.5% tryptone, and 0.05% sodium chloride.

[0013] In the present application, in step (1), the fermentation broth contains 2% glucose, 1% glycerol, 3% soybean powder, and 1% glutamic acid.

[0014] In the present application, in step (2), the acid may be any one or more of hydrochloric acid, sulfuric acid, and acetic acid.

[0015] In the present application, in step (3), the ratio of the mass of the diatomaceous earth to the volume of the Bacillus subtilis fermentation broth after pH adjustment can be 1 g / ml-3 g / ml.

[0016] In the present application, in step (3), sedimentation can be performed at 0°C-35°C.

[0017] In the present application, in step (3), the sedimentation can take place for 6-12 hours.

[0018] In the present application, in step (3), sedimentation can be carried out at 0°C-35°C for 6-12 hours.

[0019] In the present application, in step (4), the membrane may include a ceramic membrane and / or a hollow fiber membrane, wherein the pore size of the membrane is 0.1 μm-1 μm.

[0020] The present application also provides a multi-dimensional fermented essence powder, which comprises, by mass fraction: 50%-95% polyglutamic acid, 5%-50% fructan LEVEN, 100 ppm-300 ppm vitamin K2, 10 ppm-100 ppm folic acid, ppm represents the unit of concentration, one millionth of a gram, wherein the molecular weight of the polyglutamic acid is 400 D-3,000,000 D.

[0021] In the present application, the multi-dimensional fermented essence powder contains 50%-80% polyglutamic acid, 5%-20% fructan LEVAN, 100 ppm-300 ppm vitamin K2, and 10 ppm-100 ppm folic acid by mass fraction, ppm represents the unit of concentration, one millionth of a gram, wherein the molecular weight of the polyglutamic acid is 400 D-1200,000 D.

[0022] The present application also provides a method for preparing the multidimensional fermentation essence powder, which comprises: subjecting the multidimensional fermentation essence liquid to alcohol precipitation and drying to obtain the multidimensional fermentation essence powder.

[0023] In the present application, the alcohol may be selected from any one or more of methanol, ethanol, and propanol, and the volume of the alcohol may be 50%-80% of the volume of the multi-dimensional fermentation essence.

[0024] The present application also provides a cosmetic comprising the multidimensional fermentation essence liquid or the multidimensional fermentation essence powder.

[0025] The present application also provides a facial mask comprising the multi-dimensional fermentation essence liquid or the multi-dimensional fermentation essence powder.

[0026] The multi-dimensional fermented essence and multi-dimensional fermented essence powder of the present application can obviously increase the moisture content of the skin, reduce the degree of spot fading, tighten the texture, and increase the visibility of pores, and have the effects of whitening, moisturizing, anti-wrinkle and anti-freckle.

[0027] Other features and advantages of the present application will be described in the following description, and partly become apparent from the description, or be understood by implementing the present application. Other advantages of the present application can be realized and obtained by the schemes described in the description. DETAILED DESCRIPTION

[0028] In order to make the purpose, technical solution and advantages of the present application more clear, the embodiments of the present application will be described in detail below. It should be noted that the embodiments and features in the embodiments of the present application can be combined with each other arbitrarily without conflict.

[0029] Example 1 Preparation of Bacillus subtilis fermentation broth: Bacillus subtilis natto was cultured on LB solid slant medium for 24 hours, and then prepared into bacterial suspension with 5L sterile water. 5L Bacillus subtilis suspension was put into a seed tank filled with 160LB liquid for expansion culture for 3.5h. After the seed liquid concentration reached a bacterial activity of OD600 between 1.2-2.2, it was put into a fermentation tank filled with 2T fermentation liquid 1.4T (the fermentation liquid contained 2% glucose, 1% glycerol, 3% soybean powder, 1% glutamic acid) for fermentation for 23h. During this period, feed was supplemented to keep the bacteria in a highly active state during the fermentation process and to make the metabolites in the fermentation liquid accumulate at a high concentration.

[0030] Preparation of multidimensional fermentation essence: The pH of the Bacillus subtilis fermentation liquid was adjusted to 3.90 with 6N hydrochloric acid, 2% diatomaceous earth was added and stirred evenly, and then settled for 12 hours. The supernatant was taken and passed through a ceramic membrane to remove protein, salt and pigment, and concentrated to obtain the essence.

[0031] Example 2 Preparation of multidimensional fermentation essence powder: The pH of the Bacillus subtilis fermentation liquid was adjusted to 3.90 with 6N sulfuric acid, 1.5% diatomaceous earth was added, and the mixture was stirred evenly and allowed to settle for 12 hours. The supernatant was passed through a ceramic membrane to remove protein, salt and pigment, and concentrated to obtain an essence liquid. The essence liquid was then precipitated with 50% ethanol and vacuum dried to obtain a multi-dimensional fermentation essence powder.

[0032] Performance Testing The essence was added to the auxiliary materials to make a mask, and the essence powder was dissolved in water at a mass ratio of 4%-10%, emulsified, filled, packaged, and made into a mask. The results of instrument testing and human face experience are shown in Tables 1 and 2. Testing method: On the 7th and 14th days of using the product of the application, eight major items such as spots, ultraviolet spots, brown spots, red areas, wrinkles, sebum secretion, number of pores, and skin smoothness were measured using a skin quality tester (Canfield, USA, VISIA); and then the multi-functional skin quality tester black and red pigment probe (Mexameter ®, MX18) and elasticity probe (CT-MPA580) were used to measure the skin melanin, red pigment, elasticity, and moisturizing degree, a total of 12 items were tested. Before the test, the subjects washed their faces and rested for 20 minutes before entering the laboratory. The skin values ​​of the subjects' faces were tested with a skin analyzer, a multifunctional skin analyzer, and a multifunctional skin analyzer (the test areas were: right cheek, left cheek, and forehead. Each subject was required to apply the product of this application on the face in the morning and evening during the experiment. On the 7th and 14th days after use, the subjects returned to the laboratory twice to conduct skin tests after using the product of this application. Environmental interference was eliminated during the test). The test was conducted using a skin analyzer. The tester first removed makeup and cleansed the face. When the whole face was bare, the skin condition before using the mask was measured (skin moisture, spot amount, firmness, and pore visibility, among which moisture was used to test the moisturizing ability; spot amount was used to test the spot removal ability; firmness was used to test the skin firming ability; and pore visibility was used to test the pore shrinking ability). Then, the essence was applied to the face. After 15 minutes, the skin was tested again. The reference data obtained by comparing the before and after tests were used, and the before and after control values ​​of 20 people were averaged to obtain the above data. Therefore, there is no unit data for the amount of spots, firmness, and visibility of pores. The tester can only test the before-and-after comparison data.

[0033] The VISIA skin detector uses optical imaging, RBX and software technology to instantly measure and analyze epidermal spots, pores, wrinkles and skin texture, as well as subcutaneous blood vessels and pigmented lesions caused by ultraviolet radiation, such as porphyrin (oil), brown spots, erythema, etc., and reveals potential dangers caused by them, such as chloasma, acne, rosacea and spider veins.

[0034] Table 1 Test results of facial masks containing multi-dimensional fermented essence Number of people tested 20 people Skin hydration An average increase of 16% Spot lightening degree Reduces spots by 5% on average Texture firmness Increases firmness by 10% on average Pore ​​Visibility Reduces pore size by 9% on average Table 2 Test results of facial masks containing multi-dimensional fermented essence powder Number of people tested 20 people Skin hydration An average increase of 15% Spot lightening degree Reduces spots by 6% on average Texture firmness An average of 9% firmness increase Pore ​​Visibility Reduces pore size by 9% on average It can be seen from Tables 1 and 2 that after using the facial mask containing the multi-dimensional fermentation essence or multi-dimensional essence powder of the present application, the skin moisture content, spot fading degree, texture firmness, and pore visibility are all significantly increased, indicating that the multi-dimensional fermentation liquid and multi-dimensional fermentation powder of the present application have the effects of whitening, moisturizing, anti-wrinkle and anti-freckle.

[0035] Although the embodiments disclosed in this application are as above, the contents described are only embodiments adopted to facilitate understanding of this application and are not intended to limit this application. Any technician in the field to which this application belongs can make any modifications and changes in the form and details of implementation without departing from the spirit and scope disclosed in this application, but the scope of patent protection of this application shall still be based on the scope defined in the attached claims.

Claims

1. A multi-dimensional fermented essence, comprising, by mass fraction, 0.1%-30% polyglutamic acid, 0.01%-15.6% fructan LEVAN, 0.2 ppm-94 ppm vitamin K2, and 0.02 ppm-32 ppm folic acid, wherein ppm represents the unit of concentration, one millionth of a gram, wherein the molecular weight of the polyglutamic acid is 400D-3,000,000D.

2. The multidimensional fermentation essence according to claim 1, comprising, by mass fraction, 0.1%-30% polyglutamic acid, 4%-10% levan LEVE, 40 ppm-94 ppm vitamin K2, and 0.02 ppm-32 ppm folic acid, where ppm represents the unit of concentration, one millionth of a gram, wherein the molecular weight of the polyglutamic acid is 2,000-1,000,000D.

3. A method for preparing the multidimensional fermentation essence according to claim 1 or 2, the preparation method comprising: (1) Preparation of Bacillus subtilis fermentation broth: Put a Bacillus subtilis suspension with a mass concentration of 5%-20% into a seed tank filled with LB solution for expansion. After the seed liquid concentration reaches a bacterial activity of OD600 between 1.2 and 2.2, put it into a fermentation tank filled with fermentation broth and ferment it at 35° C.-50° C. for 10 hours to 48 hours to obtain the Bacillus subtilis fermentation broth; (2) adjusting the pH of the Bacillus subtilis fermentation broth to 3.9-4.5 with an acid; (3) Add diatomaceous earth, let it settle, and discard the precipitate; (4) The supernatant obtained in step (3) is filtered through a membrane to remove protein and salt, and then washed and decolorized with water to obtain the multi-dimensional fermentation essence.

4. The preparation method according to claim 3, wherein In step (1), the Bacillus subtilis is isolated from natto and is a wild-type strain of Bacillus subtilis natto ST188, CGMCC No.8400; Optionally, the LB liquid is Luria-Bertani liquid medium: calculated by mass volume ratio g / 100 ml water, comprising 0.1% glucose, 1% yeast powder, 0.5% tryptone, and 0.05% sodium chloride; Optionally, the fermentation broth contains 2% glucose, 1% glycerol, 3% soybean flour, and 1% glutamic acid.

5. The preparation method according to claim 3, wherein: In step (2), the acid is any one or more of hydrochloric acid, sulfuric acid, and acetic acid.

6. The preparation method according to claim 3, wherein: In step (3), the ratio of the mass of the diatomaceous earth to the volume of the Bacillus subtilis fermentation broth after pH adjustment is 1 g / ml to 3 g / ml; Optionally, in step (3), sedimentation is carried out at 0°C to 35°C for 6 to 12 hours; Optionally, in step (4), the membrane comprises a ceramic membrane and / or a hollow fiber membrane, wherein the pore size of the membrane is 0.1 μm-1 μm.

7. A multi-dimensional fermented essence powder, comprising, by mass fraction: 50%-95% polyglutamic acid, 5%-50% fructan LEVAN, 100 ppm-300 ppm vitamin K2, 10 ppm-100 ppm folic acid, ppm represents the unit of concentration, one millionth of a gram, wherein the molecular weight of the polyglutamic acid is 400 D-3,000,000 D.

8. The multi-dimensional fermented essence powder according to claim 7 comprises, by mass fraction: 50%-80% polyglutamic acid, 5%-20% fructan LEVAN, 100 ppm-300 ppm vitamin K2, 10 ppm-100 ppm folic acid, ppm represents the unit of concentration, one millionth of a gram, wherein the molecular weight of the polyglutamic acid is 400 D-1200,000 D.

9. A method for preparing the multi-dimensional fermentation essence powder according to claim 7 or 8, the preparation method comprising: The multidimensional fermentation essence liquid according to claim 1 is subjected to alcohol precipitation and drying to obtain the multidimensional fermentation essence powder; Optionally, the alcohol used in the alcohol precipitation is selected from any one or more of methanol, ethanol, and propanol, and the volume of the alcohol is 50%-80% of the volume of the multi-dimensional fermentation essence.

10. A cosmetic comprising the multidimensional fermentation essence liquid according to claim 1 or 2 or the multidimensional fermentation essence powder according to claim 7 or 8.