Cactus fermentation product and application thereof

Abstract

The invention relates to a cactus fermentation product and application thereof, and belongs to the technical field of fermentation. Lactobacillus liquid, streptomyces liquid, saccharomycetes liquid and bifidobacterium liquid are used for preparing mixed bacterial liquid, the cactus raw material is subjected to fermentation extraction, the prepared cactus fermentation product serves as a raw material for subsequent cosmetic preparation, and components, beneficial to skin, in the cactus fermentation product comprise polysaccharide; by mixing and combining various microorganisms, the extraction efficiency of fermentation extraction on the cactus raw material is improved, the content of beneficial components in a fermented product is improved, and the biological activity and bioavailability of the beneficial components are improved. The method has the characteristics of high fermentation efficiency and high content of beneficial components.

Description

Technical Field

[0001] The invention belongs to the technical field of fermentation and relates to a cactus fermentation product and application thereof. Background Art

[0002] Cactus stems have significant application value in the field of cosmetics. Their extracts are rich in nutrients such as protein, minerals, cellulose and vitamin C, and can be used for whitening, anti-allergy, moisturizing and anti-oxidation. Cactus fruit is the fruit of the cactus plant. The antioxidant capacity of cactus fruit is high. Studies have shown that cactus fruit extract has the effects of accelerating wound healing, making the skin soft and delicate, preventing skin dryness, and reducing wrinkles.

[0003] Cactus fermented products refer to cactus materials processed through a fermentation process, which usually involves fermenting cactus raw materials with microorganisms to produce products with specific functional ingredients. Cactus fermented products have a wide range of application value, especially in the fields of cosmetics, food and health products.

[0004] Currently, the cactus fermented products used in cosmetics on the market have low content of beneficial ingredients, low fermentation efficiency, and sometimes chemical ingredients are added during the fermentation process, which aggravates the loss of active ingredients. There is still much room for improvement. Summary of the invention

[0005] The invention aims to provide a cactus fermentation product and application thereof, which has the characteristics of high fermentation efficiency and high content of beneficial components.

[0006] The purpose of the present invention can be achieved through the following technical solutions:

[0007] A cactus fermented product, characterized in that it is obtained by the following preparation method:

[0008] S1: Take cactus stems and cactus fruits, remove thorns, wash with clean water, cut into small pieces, add deionized water, soak for 2 to 4 hours, and obtain cactus pieces;

[0009] S2: putting the cactus pieces into a rubber mill for crushing and grinding to obtain a cactus homogenate;

[0010] S3: inoculating the mixed bacterial liquid into the cactus homogenate at an inoculation amount of 8-10% of the mass of the cactus homogenate, and fermenting and culturing at a constant temperature with shaking, and obtaining a fermentation product A after the fermentation is completed;

[0011] S4: centrifuging the fermentation product A, taking the supernatant, and sterilizing it under high pressure at a temperature of 110-125° C. for 20 min to obtain supernatant A;

[0012] S5: Filter the supernatant A by suction, and collect the filtrate to obtain the cactus fermentation product.

[0013] Preferably, in S3, the mixed bacterial solution includes a lactobacillus bacterial solution, a streptomyces bacterial solution, a yeast bacterial solution and a bifidobacterium bacterial solution in a volume ratio of 5-6:2-3:2-3:2-3, and the concentration of each bacterial species is 10 5 ~10 8 CFU / mL.

[0014] Preferably, the mixed bacterial liquid is obtained by the following preparation method:

[0015] a: Pick single clones of Lactobacillus, Streptomyces, Saccharomyces and Bifidobacterium, inoculate them into liquid culture medium respectively, expand the culture and obtain bacterial mother liquid;

[0016] b: Inoculate the bacterial mother solution into fresh liquid culture medium at an inoculation rate of 1-5%, and culture until the bacterial count reaches 10 5 ~10 8 CFU / mL, bacterial solution was obtained;

[0017] c: Mix the four bacterial solutions according to the above proportions to obtain a mixed bacterial solution.

[0018] Preferably, in said a, the liquid culture medium contains the following components per liter: 10 g / L beef extract, 5 g / L yeast extract, 20 g / L soybean powder, 20 g / L glucose, 2 g / L diammonium hydrogen citrate, 1 g / L Tween 80, 0.25 g / L MnSO4·1H2O, 0.58 g / L MgSO4·7H2O, 2 g / L dipotassium hydrogen phosphate and 5 g / L sodium acetate, the balance being water, and pH=6.0-6.6.

[0019] Preferably, in S3, the culture temperature is 30-37° C., the shaking speed is 140-160 rpm, and the culture time is 24-72 h.

[0020] Preferably, in S4, the centrifugal speed is set to 4000-6000 r / min.

[0021] Preferably, the pore size of the filtration membrane is in the range of 0.4 to 10 μm.

[0022] Preferably, it is used in the preparation of cosmetics.

[0023] The present invention uses a mixed bacterial liquid prepared by using lactobacillus liquid, streptomyces liquid, yeast liquid and bifidobacterium liquid to ferment and extract a cactus raw material, so as to obtain a cactus fermented product as a raw material for subsequent preparation of cosmetics. The ingredients in the cactus fermented product that are beneficial to the skin include polysaccharides such as pectin and cellulose, amino acids, vitamins such as vitamin C and E, and minerals. Through the mixed use of multiple microorganisms, the extraction efficiency of the cactus raw material by fermentation extraction is improved, the content of the beneficial ingredients in the fermented product is improved, the biological activity and bioavailability of the beneficial ingredients are increased, and compared with other extraction methods such as chemical extraction, the biological fermentation extraction is more gentle, better retains the active ingredients, reduces the loss of the active ingredients that may be caused by chemical treatment, and is also helpful to reduce skin irritation, so the method can be widely used in the field of cosmetics.

[0024] Polysaccharides, amino acids and vitamins in cactus can be extracted through lactobacillus fermentation. Polysaccharides can provide significant moisturizing effects and enhance skin hydration. Amino acids have a positive effect on skin repair and regeneration. Vitamins C and E help resist free radical damage, help resist oxidative stress, and slow down the aging process of the skin. This fermentation process can promote the transformation and enhancement of active ingredients in cactus, and improve its moisturizing, repair and antioxidant capabilities.

[0025] The combined fermentation of bifidobacteria and lactobacilli can improve the bioavailability of polysaccharides and amino acids, and increase the skin repair and moisturizing effects.

[0026] Lactobacillus produces lactic acid through fermentation, which lowers the pH value of the environment and makes it more acidic. Among the enzymes produced by Streptomyces and yeast, several types are particularly suitable for combining with the acidic environment produced by lactobacillus fermentation to help decompose the complex components in cactus. Streptomyces can produce cellulase, pectinase and β-glucosidase, and yeast can produce cellulase and β-glucosidase. The acidic environment produced by lactobacillus fermentation can activate these enzymes, enhance their activity, and decompose the complex polysaccharides in cactus more effectively and thoroughly, so that the polysaccharides are degraded into more easily usable substances. For example, cellulase and β-glucosidase can decompose cellulose and pectin, the main components of cactus cell walls, and release antioxidants such as flavonoids, polyphenols and vitamin C. The acidic environment is conducive to this process, improves the extraction efficiency of antioxidants, and maintains their activity and stability to ensure their effectiveness in cosmetic products.

[0027] The fermentation product of yeast, ethanol, has antibacterial effects and can inhibit the growth of some harmful microorganisms. During joint fermentation, this antibacterial effect helps reduce the interference of competing microorganisms and improve the fermentation environment, which is beneficial to the growth of combined strains and the fermentation process, and improves the extraction efficiency of cactus raw materials by fermentation extraction.

[0028] The beneficial effects of the present invention are:

[0029] The present invention uses a combination of multiple microorganisms to improve the extraction efficiency of fermentation extraction on cactus raw materials, increase the content of beneficial ingredients in the fermented product, and increase the biological activity and bioavailability of the beneficial ingredients. Compared with other extraction methods, such as chemical extraction, biological fermentation extraction is milder and better retains active ingredients, reduces the loss of active ingredients that may be caused by chemical treatment, and also helps to reduce skin irritation. Therefore, the present invention can be widely used in the field of cosmetics.

[0030] The invention uses lactobacillus liquid, streptomyces liquid, yeast liquid and bifidobacterium liquid to prepare a mixed liquid for fermentation and extraction of cactus raw materials. The combined fermentation of bifidobacteria and lactobacillus can improve the bioavailability of polysaccharides and amino acids, and increase the skin repair and moisturizing effects. The enzymes produced by streptomyces and yeast are combined with the acidic environment produced by lactobacillus to promote the degradation of complex polysaccharides, improve the extraction efficiency of antioxidant substances, and maintain their activity and stability to ensure their effects in cosmetic products. The fermentation product ethanol of yeast has an antibacterial effect and can inhibit the growth of some harmful microorganisms. During the combined fermentation, the antibacterial effect helps to reduce the interference of competing microorganisms and improve the fermentation environment, thereby facilitating the growth of the combined strains and the fermentation process, and improving the extraction efficiency of the cactus raw materials by fermentation extraction. DETAILED DESCRIPTION

[0031] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation methods, structures, features and effects of the present invention are described in detail below in combination with the embodiments.

[0032] The mixed bacterial solution used in each embodiment and comparative example of the present invention is obtained by the following preparation method:

[0033] a: picking up single clones of lactobacillus, streptomyces, yeast and bifidobacterium, and inoculating them into liquid culture medium respectively according to the actual strains used in each embodiment and comparative example, and expanding the culture to obtain bacterial mother liquid;

[0034] b: Inoculate the mother liquid into fresh liquid culture medium at a 1% inoculum volume and culture until the number of bacteria reaches 10 8 CFU / mL, bacterial solution was obtained;

[0035] c: The bacterial solutions were mixed according to the proportions in the examples and comparative examples to obtain a mixed bacterial solution.

[0036] Embodiment 1:

[0037] S1: Take cactus stems and cactus fruits, remove thorns, wash with clean water, cut into small pieces, add deionized water, soak for 2 hours, and obtain cactus pieces;

[0038] S2: putting the cactus pieces into a rubber mill for crushing and grinding to obtain a cactus homogenate;

[0039] S3: inoculating the mixed bacterial liquid into the cactus homogenate at an inoculation amount of 10% of the mass of the cactus homogenate, and culturing the mixture under constant temperature shaking at 35° C. and a shaking speed of 150 rpm for 48 hours, and obtaining a fermentation product A after the fermentation is completed;

[0040] S4: centrifuging the fermentation product A at a centrifugal speed of 4000 r / min, taking the supernatant, and sterilizing it under high pressure at 121° C. for 20 min to obtain supernatant A;

[0041] S5: The supernatant A is filtered through a filter membrane with a pore size of 0.5 μm, and the filtrate is collected to obtain the cactus fermentation product.

[0042] The mixed bacterial solution includes lactobacillus bacterial solution, streptomyces bacterial solution, yeast bacterial solution and bifidobacterium bacterial solution in a volume ratio of 5:2:2:2.

[0043] Embodiment 2:

[0044] S1: Take cactus stems and cactus fruits, remove thorns, wash with clean water, cut into small pieces, add deionized water, soak for 2 hours, and obtain cactus pieces;

[0045] S2: putting the cactus pieces into a rubber mill for crushing and grinding to obtain a cactus homogenate;

[0046] S3: inoculating the mixed bacterial liquid into the cactus homogenate at an inoculation amount of 10% of the mass of the cactus homogenate, and culturing the mixture under constant temperature shaking at 35° C. and a shaking speed of 150 rpm for 48 hours, and obtaining a fermentation product A after the fermentation is completed;

[0047] S4: centrifuging the fermentation product A at a centrifugal speed of 4000 r / min, taking the supernatant, and sterilizing it under high pressure at 121° C. for 20 min to obtain supernatant A;

[0048] S5: The supernatant A is filtered through a filter membrane with a pore size of 0.5 μm, and the filtrate is collected to obtain the cactus fermentation product.

[0049] The mixed bacterial solution includes lactobacillus bacterial solution, streptomyces bacterial solution, yeast bacterial solution and bifidobacterium bacterial solution in a volume ratio of 6:3:3:2.

[0050] Embodiment 3:

[0051] S1: Take cactus stems and cactus fruits, remove thorns, wash with clean water, cut into small pieces, add deionized water, soak for 2 hours, and obtain cactus pieces;

[0052] S2: putting the cactus pieces into a rubber mill for crushing and grinding to obtain a cactus homogenate;

[0053] S3: inoculating the mixed bacterial liquid into the cactus homogenate at an inoculation amount of 8% of the mass of the cactus homogenate, and culturing the mixture under constant temperature shaking at 35° C. and a shaking speed of 150 rpm for 48 hours, and obtaining a fermentation product A after the fermentation is completed;

[0054] S4: centrifuging the fermentation product A at a centrifugal speed of 4000 r / min, taking the supernatant, and sterilizing it under high pressure at 121° C. for 20 min to obtain supernatant A;

[0055] S5: The supernatant A is filtered through a filter membrane with a pore size of 0.5 μm, and the filtrate is collected to obtain the cactus fermentation product.

[0056] The mixed bacterial liquid includes lactobacillus bacterial liquid, streptomyces bacterial liquid, yeast bacterial liquid and bifidobacterium bacterial liquid in a volume ratio of 5:2:3:2.

[0057] Example 4

[0058] S1: Take cactus stems and cactus fruits, remove thorns, wash with clean water, cut into small pieces, add deionized water, soak for 2 hours, and obtain cactus pieces;

[0059] S2: putting the cactus pieces into a rubber mill for crushing and grinding to obtain a cactus homogenate;

[0060] S3: inoculating the mixed bacterial liquid into the cactus homogenate at an inoculation amount of 8% of the mass of the cactus homogenate, and culturing the mixture under constant temperature shaking at 35° C. and a shaking speed of 150 rpm for 48 hours, and obtaining a fermentation product A after the fermentation is completed;

[0061] S4: centrifuging the fermentation product A at a centrifugal speed of 4000 r / min, taking the supernatant, and sterilizing it under high pressure at 121° C. for 20 min to obtain supernatant A;

[0062] S5: The supernatant A is filtered through a filter membrane with a pore size of 0.5 μm, and the filtrate is collected to obtain the cactus fermentation product.

[0063] The mixed bacterial solution includes lactobacillus bacterial solution, streptomyces bacterial solution, yeast bacterial solution and bifidobacterium bacterial solution in a volume ratio of 5:2:2:3.

[0064] Comparative Example 1

[0065] In this comparative example, only lactobacillus liquid was added, and other steps were consistent with those in Example 1.

[0066] Comparative Example 2

[0067] In this comparative example, only the lactobacillus culture liquid and the bifidobacterium culture liquid were added in a volume ratio of 5:2, and the other steps were consistent with those in Example 1.

[0068] Comparative Example 3

[0069] In this comparative example, lactobacillus liquid, yeast liquid and bifidobacterium liquid were added in a volume ratio of 5:3:3, and the other steps were consistent with Example 1.

[0070] Comparative Example 4

[0071] In this comparative example, a Lactobacillus solution, a Streptomyces solution and a Bifidobacterium solution were added in a volume ratio of 5:3:3, and other steps were consistent with Example 1.

[0072] Example 5

[0073] Determination of total flavonoids content

[0074] The flavonoid mother nucleus contains alkaline oxygen atoms, which generally have phenolic hydroxyl groups and can produce yellow complexes with aluminum ions. The addition of sodium nitrite and sodium hydroxide makes it red in alkaline solution. Rutin is used as a reference substance and aluminum chloride is used as a colorimetric developer for flavonoid colorimetric determination. The absorbance is linearly related to the concentration of rutin. The total flavonoid content was determined by spectrophotometry.

[0075] Use a pipette to take 1 mL of the fermented product, add 0.3 mL of 5% sodium nitrite solution to each, vortex to mix, let stand for 6 min, then add 0.3 mL of aluminum chloride solution, vortex to mix, let stand for another 6 min, then add 4 mL of 4% sodium hydroxide solution, vortex to mix, let stand for 15 min, measure the absorbance at 510 nm and enter into the following formula, set up 3 parallel experiments for each group to calculate the average value.

[0076] The standard curve of rutin is y=0.0056x+0.0123, R 2 =0.9998.

[0077] Table 1 shows the total flavonoid content of each fermentation product in Examples 1 to 4 and Comparative Examples 1 and 2.

[0078] Table 1: Total flavonoid content of each fermentation product

[0079] Total flavonoids content (mgRE / gDW) Example 1 28.34 Example 2 35.23 Example 3 33.67 Example 4 29.88 Comparative Example 1 16.98 Comparative Example 2 20.78 Comparative Example 3 22.23 Comparative Example 4 23.45

[0080] The results showed that when the four strains were added simultaneously for joint fermentation, the total flavonoids content of the fermented product was higher, and the fermentation efficiency was highest when the four strains were fermented together.

[0081] Example 6

[0082] Determination of total phenolic content

[0083] The total phenol content was determined using the Folin-phenol method. In an alkaline solution, phenolic compounds were oxidized by the Folin-phenol reagent to generate blue compounds. The depth of the blue color was proportional to the number of phenolic groups. Gallic acid was used as a reference.

[0084] Take 1 mL of the fermented product with a pipette, add 5 mL of Folin reagent and 4 mL of 75 g / L sodium carbonate solution, vortex and mix well, let stand in a 40°C constant temperature water bath for 30 min, measure the absorbance at 765 nm and enter it into the following formula. Set up 3 parallel experiments for each group to calculate the average value.

[0085] The standard curve of gallic acid is y=0.0109x+0.045, R 2 =0.9996.

[0086] Table 2 shows the total phenol content of each fermentation product in Examples 1 to 4 and Comparative Examples 1 and 2.

[0087] Table 2: Total phenolic content of each fermentation product

[0088] Total phenolic content (mgGAE / gDW) Example 1 26.55 Example 2 30.46 Example 3 29.99 Example 4 27.41 Comparative Example 1 12.78 Comparative Example 2 19.56 Comparative Example 3 20.33 Comparative Example 4 20.98

[0089] The results showed that when the four strains were added simultaneously for joint fermentation, the total phenol content of the fermentation product was higher, and the fermentation efficiency was highest when the four strains were fermented together.

[0090] Example 5

[0091] Determination of antioxidant activity by DPPH method

[0092] DPPH, also known as 1,1-diphenyl-2-trinitrophenylhydrazine, is a stable free radical in organic solvents. Its alcohol solution is purple and can accept electrons or hydrogen atoms. When antioxidants are added, the number of free radicals decreases, the absorbance value decreases, and the color of the solution becomes lighter. The decrease in absorbance level indicates an increase in antioxidant activity, thereby evaluating the antioxidant capacity of the test sample.

[0093] Take 100 μL of the fermented product with a pipette, add 400 μL of DPPH ethanol solution (200 μg / mL), mix well, react at room temperature in the dark for 30 min, and measure the absorbance value A1 at 517 nm;

[0094] Take 100 μL of the fermented product with a pipette, add 400 μL of DPPH ethanol solution (200 μg / mL), mix well, and directly measure its absorbance value A2 at 517 nm;

[0095] Take 500 μL of DPPH ethanol solution (200 μg / mL) with a pipette and measure its absorbance value A0 at 517 nm.

[0096] Each group was set up with 3 parallel experiments to calculate the average value. The calculation method is as follows:

[0097] Clearance % = [1-(A1-A2)] / A0×100

[0098] Table 3 shows the test results of the antioxidant capacity of each fermentation product in Examples 1 to 4 and Comparative Examples 1 and 2.

[0099] Table 3: Antioxidant capacity test results of various fermentations

[0100] Average clearance rate (%) Example 1 68.74 Example 2 75.76 Example 3 72.45 Example 4 69.13 Comparative Example 1 32.67 Comparative Example 2 45.69 Comparative Example 3 55.02 Comparative Example 4 58.99

[0101] The results showed that when four strains of bacteria were added simultaneously for joint fermentation, the fermented product had higher antioxidant properties.

[0102] When the present invention is used:

[0103] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A cactus fermentation product, characterized in that: Obtained by the following preparation method: S1: Take cactus stems and cactus fruits, remove thorns, wash with clean water, cut into small pieces, add deionized water, soak for 2 to 4 hours, and obtain cactus pieces; S2: putting the cactus pieces into a rubber mill for crushing and grinding to obtain a cactus homogenate; S3: inoculating the mixed bacterial liquid into the cactus homogenate at an inoculation amount of 8-10% of the mass of the cactus homogenate, and fermenting and culturing at a constant temperature with shaking, and obtaining a fermentation product A after the fermentation is completed; S4: centrifuging the fermentation product A, taking the supernatant, and sterilizing it under high pressure at a temperature of 110-125° C. for 20 min to obtain supernatant A; S5: Filter the supernatant A by suction, and collect the filtrate to obtain the cactus fermentation product.

2. The cactus fermented product according to claim 1, characterized in that: In S3, the mixed bacterial solution includes a lactobacillus bacterial solution, a streptomyces bacterial solution, a yeast bacterial solution and a bifidobacterium bacterial solution in a volume ratio of 5 to 6: 2 to 3: 2 to 3: 2 to 3, and the concentration of each bacterial species is 10 5 ~10 8 CFU / mL.

3. The cactus fermented product according to claim 2, characterized in that: The mixed bacterial solution is obtained by the following preparation method: a: Pick single clones of Lactobacillus, Streptomyces, Saccharomyces and Bifidobacterium, inoculate them into liquid culture medium respectively, expand the culture and obtain bacterial mother liquid; b: Inoculate the bacterial mother solution into fresh liquid culture medium at an inoculation rate of 1-5%, and culture until the bacterial count reaches 10 5 ~10 8 CFU / mL, bacterial solution was obtained; c: Mix the four bacterial solutions according to the above proportions to obtain a mixed bacterial solution.

4. The cactus fermented product according to claim 3, characterized in that: In the a, the liquid culture medium contains the following components per liter: 10 g / L beef extract, 5 g / L yeast extract, 20 g / L soybean powder, 20 g / L glucose, 2 g / L diammonium hydrogen citrate, 1 g / L Tween 80, 0.25 g / L MnSO4·1H2O, 0.58 g / L MgSO4·7H2O, 2 g / L dipotassium hydrogen phosphate and 5 g / L sodium acetate, the balance is water, and the pH is 6.0-6.

6.

5. The cactus fermented product according to claim 1, characterized in that: In S3, the culture temperature is 30-37° C., the shaking speed is 140-160 rpm, and the culture time is 24-72 h.

6. The cactus fermented product according to claim 1, characterized in that: In S4, the centrifugal speed is set to 4000-6000 r / min.

7. The cactus fermented product according to claim 1, characterized in that: The pore size of the suction filtration membrane ranges from 0.4 to 10 μm.

8. The cactus fermented product according to any one of claims 1 to 7, characterized in that: Used in the preparation of cosmetics.