A beauty - enhancing beverage fermented with probiotics containing Taizui and its preparation method

Abstract

The present invention provides a beauty - enhancing drink fermented with probiotics containing Ganoderma lucidum mycelium and its preparation method, belonging to the technical field of drinks. Chinese wolfberry, houttuynia cordata, angelica dahurica and Chinese angelica are fermented and separated by lactic acid bacteria to obtain Chinese medicine - fermented sugar and fermented extract. After the protein peptide obtained from Nostoc commune Vaucher undergoes the Maillard reaction with the Chinese medicine - fermented sugar, it chelates calcium and magnesium ions to obtain a Maillard reaction product chelating Ca / Mg, which is mixed with a fermented extract of Ganoderma lucidum rich in selenium, a fermented extract and trehalose, added to purified water, sterilized, and filled to prepare the beauty - enhancing drink fermented with probiotics containing Ganoderma lucidum mycelium. The beauty - enhancing drink fermented with probiotics containing Ganoderma lucidum mycelium prepared by the present invention is a compound drink, which has good antioxidant, anti - inflammatory, beauty - enhancing and anti - aging effects, has a good taste, a long shelf life, is convenient for storage, is deeply loved by consumers, and has broad application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of beverages, and in particular to a whitening beverage containing probiotics containing Tai Sui bacteria fermentation and a preparation method thereof. Background Art

[0002] With the improvement of modern living standards, people pay more and more attention to their external image, using a good image to increase their self-confidence in various activities and occasions. At the same time, under a beautiful appearance, they pursue skin health more, and show their pursuit of high-end life with healthy skin and a good image.

[0003] Skin color is mainly determined by the number of melanocytes in the skin and the melanin content of the cells. If the proportion of melanocytes is small and the melanin content is low, the skin will appear white. If the proportion of melanocytes is large and the melanin content is high, the skin will appear dark. In melanocytes, tyrosine is oxidized into dopa and dopaquinone under the catalytic action of tyrosinase, and dopaquinone is further converted into dopachrome. Under the catalytic action of DHI enzyme, dopachrome interconvertase and DHICA oxidase, dopachrome gradually forms melanin.

[0004] Ganoderma lucidum, also known as Tai Sui, is a strange-looking organism with strong vitality. It was later confirmed to be a large slime mold complex composed of various bacteria, fungi and their metabolites. Ganoderma lucidum has a long history in China. There is also a folk rumor that "eating it can make you live forever". It is one of the immortality elixirs that many emperors have been searching for. However, some people believe that digging up this "strange meat"-like Tai Sui will bring bad luck. Therefore, the appearance of Tai Sui in ancient times is usually full of fantasy. In addition to folk legends, Tai Sui with historical records is mainly used as medicine. The "Compendium of Materia Medica" commented: "Ganoderma lucidum is shaped like meat and is a living thing. The white one is like fat, and the yellow one is like purple gold. They are all bright and clear like ice." Doctors often use it as an antibacterial agent after using it as medicine. They believe that it has the effect of immunomodulation and even has a certain effect in tumor treatment. At present, there is no research on the preparation of whitening drinks with active substances obtained by fermentation and extraction of Tai Sui. Summary of the invention

[0005] The purpose of the present invention is to provide a whitening beverage containing Tai Sui probiotic fermentation and a preparation method thereof. The present invention is a composite beverage with good antioxidant, anti-inflammatory, whitening and anti-aging effects, good taste, long shelf life, easy storage, deeply loved by consumers, and has broad application prospects.

[0006] The technical solution of the present invention is achieved in this way:

[0007] The present invention provides a method for preparing a fermented whitening drink with Ganoderma lucidum probiotics. Chinese wolfberry, houttuynia cordata, angelica dahurica and Chinese angelica are fermented and separated by lactic acid bacteria to obtain Chinese medicine fermented sugar and fermented extract. After the protein peptide obtained from nostoc commune undergoes the Maillard reaction with the Chinese medicine fermented sugar, calcium and magnesium ions are chelated to obtain a Maillard reaction product chelated with Ca / Mg. The Maillard reaction product chelated with Ca / Mg, the fermented extract of Ganoderma lucidum rich in selenium, the fermented extract and trehalose are mixed and added to purified water, sterilized, and filled to prepare a fermented whitening drink with Ganoderma lucidum probiotics.

[0008] As a further improvement of the present invention, the method includes the following steps:

[0009] S1. Activation of fermenting bacteria: Lactobacillus plantarum, Lactobacillus rhamnosus and selenium-rich yeast are respectively inoculated into Gause's medium for activation culture to obtain a bacterial strain seed solution.

[0010] S2. Fermentation of Chinese medicine: Chinese wolfberry, houttuynia cordata, angelica dahurica and Chinese angelica are washed, mixed, pulverized, added to water, sterilized, inoculated with the bacterial strain seed solutions of Lactobacillus plantarum and Lactobacillus rhamnosus, fermented and cultured, filtered, ethanol is added to the filtrate for precipitation, the solid is washed and dried to obtain Chinese medicine fermented sugar; the ethanol in the filtrate is recovered and dried to obtain a fermented extract.

[0011] S3. Fermentation of Ganoderma lucidum: Ganoderma lucidum is washed, minced, added to water, the bacterial strain seed solution of selenium-rich yeast is added, fermented and cultured, filtered, and the filtrate is freeze-dried to obtain a fermented extract of Ganoderma lucidum rich in selenium.

[0012] S4. Extraction of nostoc commune protein: Nostoc commune algal dry powder is added to water, cooled with liquid nitrogen, thawed at room temperature, extracted by microwave heating, filtered, ethanol is added to the filtrate for precipitation, filtered, the ethanol in the filtrate is recovered and freeze-dried to obtain nostoc commune protein peptide.

[0013] S5. Maillard reaction: The Chinese medicine fermented sugar and nostoc commune protein peptide are mixed evenly, water is added, transferred to a hydrothermal reaction kettle, heated and stirred for reaction to obtain a Maillard reaction product.

[0014] S6. Chelation of metal ions: The Maillard reaction product is added to water, calcium salt and magnesium salt are added, stirred for reaction, and dried to obtain a Maillard reaction product chelated with Ca / Mg.

[0015] S7. Preparation of a fermented whitening drink with Ganoderma lucidum probiotics: The fermented extract, the fermented extract of Ganoderma lucidum rich in selenium, the Maillard reaction product chelated with Ca / Mg and trehalose are mixed and added to purified water, sterilized, and filled to prepare a fermented whitening drink with Ganoderma lucidum probiotics.

[0016] As a further improvement of the present invention, the conditions for the activation culture in step S1 are 25-35 °C, 100-200 r / min, cultured for 18-24 h, and the bacterial content of the bacterial strain seed solution is 10 8 -109 cfu / mL.

[0017] As a further improvement of the present invention, in step S2, the mass ratio of wolfberry, houttuynia cordata, angelica dahurica and Chinese angelica is 10 - 12:3 - 5:2 - 4:1 - 3. The inoculation amounts of the plant lactobacillus and the rhamnulose lactobacillus strain seed liquid are 1 - 2 v / v% and 2 - 3 v / v% respectively. The conditions for fermentation culture are anaerobic conditions, 35 - 38 °C, 100 - 200 r / min, and culture for 36 - 48 h. Ethanol is added to make the ethanol content in the system 70 - 80 wt%, and precipitation is carried out for 1 - 2 h.

[0018] As a further improvement of the present invention, in step S3, the inoculation amount of the selenium - rich yeast strain seed liquid is 1 - 2 v / v%. The conditions for fermentation culture are 25 - 30 °C, 100 - 200 r / min, and culture for 24 - 36 h.

[0019] As a further improvement of the present invention, in step S4, the solid - liquid ratio of nostoc commune algae dry powder and water is 1:7 - 10 g / mL. The power of microwave heating extraction is 1000 - 2000 W, the temperature is 45 - 55 °C, the time is 2 - 4 h. Ethanol is added to make the ethanol content in the system 70 - 80 wt%, and precipitation is carried out for 0.5 - 1 h.

[0020] As a further improvement of the present invention, in step S5, the mass ratio of traditional Chinese medicine fermented sugar, nostoc commune protein peptide and water is 12 - 15:7 - 10:20 - 30. The temperature of the heating and stirring reaction is 120 - 130 °C, and the time is 40 - 60 min.

[0021] As a further improvement of the present invention, in step S6, the mass ratio of the Maillard reaction product, calcium salt and magnesium salt is 10:1 - 2:0.5 - 1. The calcium salt is calcium chloride or calcium nitrate, the magnesium salt is magnesium chloride or magnesium nitrate, and the stirring reaction time is 20 - 40 min.

[0022] As a further improvement of the present invention, in step S7, the mass ratio of the fermentation extract, selenium - rich ganoderma lucidum fermentation extract, Maillard reaction product chelated with Ca / Mg, trehalose and purified water is 10 - 12:8 - 10:4 - 7:2 - 3:120 - 150.

[0023] The present invention further protects a ganoderma lucidum probiotic fermentation whitening drink prepared by the above - mentioned preparation method.

[0024] The present invention has the following beneficial effects:

[0025] The depth of human skin color mainly depends on the content and distribution of melanin. Tyrosine is hydroxylated to dopa by tyrosinase and then forms dopaquinone, and melanin is formed through the action of DHI oxidase and DHICA oxidase. Tyrosinase, as the key enzyme and rate-limiting enzyme for melanin synthesis in organisms, is a copper-containing oxidoreductase. In the process of its catalyzing melanin synthesis, the presence of oxygen is a necessary condition. Therefore, antioxidant substances can reduce melanin synthesis by blocking or weakening the activity of tyrosinase. Substances with both antioxidant and tyrosinase inhibitory activities can combine the effects of both to better reduce melanin synthesis. Secondly, antioxidant can also reduce free radicals generated by various exogenous factors, thereby avoiding the oxidation of collagen fibers in the skin (showing yellow) and resulting in a yellowish skin tone.

[0026] In the present invention, wolfberry has good antioxidant and whitening effects; the representative active ingredient tannic acid substances in houttuynia cordata extract have functions such as whitening, moisturizing, antioxidant, and anti-inflammatory; the volatile oils and coumarins in angelica dahurica can effectively inhibit the synthesis of intracellular melanin and the activity of tyrosinase; the active ingredients of angelica sinensis, such as volatile oils, phenolic compounds, coumarin compounds, terpene compounds, cis-apigenin, etc., all show obvious inhibitory effects on tyrosinase; the combination of the four herbs can strengthen the spleen and stomach, regulate qi movement, promote blood circulation and whiten the skin. However, directly decocting the traditional Chinese medicine components has problems such as incomplete extraction, cumbersome process, easy mildew after long-term storage, large volume for taking, and inconvenient to carry. The present invention uses lactic acid bacteria for fermentation, which not only efficiently extracts, promotes the rupture of plant cell walls, but also produces more new substances with better activity during fermentation. At the same time, the fermentation products of lactic acid bacteria mainly include organic acids, exopolysaccharides, polypeptides, bacteriocins, etc., which have antioxidant effects, can eliminate free radicals in cells and reduce the accumulation of reactive oxygen species (ROS) in the body, greatly improving antioxidant, anti-inflammatory, and whitening activities. Moreover, the obtained fermentation products are separated to obtain polysaccharides and fermentation extracts, and the extraction method is simple, and it is convenient to carry and not easy to deteriorate after drying.

[0027] Nostoc commune is a type of alga rich in protein. At the same time, it is rich in various vitamins, such as VB1, VB2, VC, VE, β-carotene, etc. After freeze-thaw treatment, it promotes the rupture of the cell wall of Nostoc commune alga, so that under the condition of microwave heating, the dissolution of active components in the alga is promoted. After alcohol precipitation to separate polysaccharides, Nostoc commune alga protein peptides are obtained.

[0028] The Maillard reaction refers to a complex chemical reaction in which the amino group contained in an amino acid or protein reacts with the carbonyl group contained in a reducing sugar under heating conditions. In the present invention, the Maillard reaction occurs between Nostoc commune protein peptides and traditional Chinese medicine fermentation sugars under heating conditions, and the prepared products such as melanoidins have significant antioxidant activity, and other active components such as polyphenols and flavonoids have effects such as nourishing qi and blood, anti-cancer, antibacterial, preventing and treating diseases, etc., thus having excellent antioxidant, anti-aging, and whitening effects.

[0029] Calcium ions are one of the extremely common metal ions in the skin and play important roles in many efficacy categories such as skin tissue regeneration, maintaining skin integrity, strengthening the skin barrier, promoting epidermal cell proliferation and differentiation, remodeling the skin extracellular matrix, as well as whitening and soothing. Magnesium ions can promote the repair of the skin barrier. After the skin is soaked in a salt bath solution rich in magnesium ions, the skin tolerance is improved, the skin barrier function and the water hydration degree of the stratum corneum are enhanced, and the skin roughness is improved.

[0030] In the present invention, the prepared Maillard reaction product is mixed and reacted with calcium salt and magnesium salt, which greatly increases the fixation amount of metal ions, avoids the loss of metal ions after entering the human body, promotes their absorption in the intestine, and greatly increases the absorption amount of metal ions by the human body, so as to better exert the effects of whitening and improving the skin.

[0031] In the present invention, selenium-enriched yeast is used to ferment Taizishen, and the prepared selenium-enriched Taizishen fermentation extract is rich in selenium element and has good antioxidant effects and can improve the body immunity. At the same time, the complex bacteria and their products in Taizishen have good antioxidant and whitening effects and can also extend the shelf life of the drink.

[0032] The Taizishen probiotic fermentation whitening drink prepared in the present invention is a compound drink, which has good antioxidant, anti-inflammatory, whitening and anti-aging effects, has a good taste, a long shelf life and is convenient for storage, is deeply loved by consumers and has broad application prospects. Detailed implementation manners

[0033] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0034] Lactobacillus plantarum, 10 billion cfu / g, Lactobacillus rhamnosus, 10 billion cfu / g, purchased from Zhongke Jiayi Bioengineering Technology Co., Ltd.; selenium-enriched yeast, 20 billion cfu / g, purchased from Angel Yeast Co., Ltd. Nostoc commune algae dry powder, water content <5%, purchased from Hunan Yandi Bioengineering Co., Ltd.

[0035] Example 1

[0036] This example provides a preparation method of a Taizishen probiotic fermentation whitening drink, including the following steps:

[0037] S1. Activation of fermentation bacteria: Lactobacillus plantarum, Lactobacillus rhamnosus and selenium-enriched yeast were respectively inoculated into Gause's medium, cultured at 25 °C and 100 r / min for 18 - 24 h to obtain a strain seed solution with a bacterial content of 10 8 -10 9 cfu / mL;

[0038] S2. Fermentation of traditional Chinese medicine: 10 g of wolfberry, 3 g of houttuynia, 2 g of angelica dahurica and 1 g of Chinese angelica were washed, mixed, crushed, added to 500 mL of water, sterilized, inoculated with Lactobacillus plantarum and Lactobacillus rhamnosus strain seed solutions, and the inoculation amounts were 1 v / v% and 2 v / v% respectively. Under anaerobic conditions, cultured at 35 °C and 100 r / min for 36 h, filtered, ethanol was added to the filtrate until the ethanol content in the system was 70 wt%, precipitated for 1 h, the solid was washed and dried to obtain traditional Chinese medicine fermented sugar; the ethanol in the filtrate was recovered and dried to obtain a fermented extract;

[0039] S3. Fermentation of Ganoderma lucidum: 10 g of Ganoderma lucidum was washed, minced, added to 200 mL of water, added with selenium-enriched yeast strain seed solution, and the inoculation amount was 1 v / v%. Cultured at 25 °C and 100 r / min for 24 h, filtered, and the filtrate was freeze-dried to obtain a selenium-enriched Ganoderma lucidum fermented extract;

[0040] S4. Extraction of Nostoc commune protein: Nostoc commune algal dry powder was added to water, and the solid-liquid ratio was 1:7 g / mL. It was cooled with liquid nitrogen, melted at room temperature, heated to 45 °C with 1000 W microwave for 2 h, filtered, ethanol was added to the filtrate until the ethanol content in the system was 70 wt%, precipitated for 0.5 h, filtered, the filtrate was collected, the ethanol in the filtrate was recovered and freeze-dried to obtain Nostoc commune protein peptide;

[0041] S5. Maillard reaction: 12 g of traditional Chinese medicine fermented sugar and 7 g of Nostoc commune protein peptide were mixed evenly, added with 20 g of water, transferred to a hydrothermal reaction kettle, heated to 120 °C, and stirred for 40 min to obtain a Maillard reaction product;

[0042] S6. Metal ion chelation: 10 g of Maillard reaction product was added to 200 mL of water, 1 g of calcium chloride and 0.5 g of magnesium chloride were added, stirred and reacted for 20 min, and dried to obtain a Maillard reaction product chelated with Ca / Mg;

[0043] S7. Preparation of Ganoderma lucidum probiotic fermented whitening drink: 10 g of fermented extract, 8 g of selenium-enriched Ganoderma lucidum fermented extract, 4 g of Maillard reaction product chelated with Ca / Mg and 2 g of trehalose were mixed and added to 120 g of purified water, pasteurized and filled to obtain Ganoderma lucidum probiotic fermented whitening drink.

[0044] Example 2

[0045] This embodiment provides a method for preparing a fermented whitening drink with Taizui probiotics, comprising the following steps:

[0046] S1. Activation of fermenting bacteria: Lactobacillus plantarum, Lactobacillus rhamnosus, and selenium-enriched yeast are respectively inoculated into Gao's medium, cultured at 35 °C and 200 r / min for 18 - 24 h to obtain a strain seed solution with a bacterial content of 10 8 -10 9 cfu / mL;

[0047] S2. Fermentation of traditional Chinese medicine: 12 g of wolfberry, 5 g of houttuynia cordata, 4 g of angelica dahurica, and 3 g of Chinese angelica are washed, mixed, pulverized, added to 500 mL of water, sterilized, inoculated with the strain seed solutions of Lactobacillus plantarum and Lactobacillus rhamnosus, with inoculation amounts of 2 v / v% and 3 v / v% respectively. Under anaerobic conditions, cultured at 38 °C and 200 r / min for 48 h, filtered. Ethanol is added to the filtrate until the ethanol content in the system is 80 wt%, precipitated for 2 h, the solid is washed and dried to obtain fermented traditional Chinese medicine sugar; the ethanol in the filtrate is recovered and dried to obtain a fermented extract;

[0048] S3. Fermentation of Taizui: 10 g of Taizui is washed, minced, added to 200 mL of water, added with the strain seed solution of selenium-enriched yeast, with an inoculation amount of 2 v / v%, cultured at 30 °C and 200 r / min for 36 h, filtered, and the filtrate is freeze-dried to obtain a selenium-enriched Taizui fermented extract;

[0049] S4. Extraction of Nostoc commune protein: Nostoc commune algae dry powder is added to water, with a solid-liquid ratio of 1:10 g / mL, cooled with liquid nitrogen, thawed at room temperature, heated to 55 °C by 2000 W microwave for 4 h, filtered. Ethanol is added to the filtrate until the ethanol content in the system is 80 wt%, precipitated for 1 h, filtered, the filtrate is collected, the ethanol in the filtrate is recovered and freeze-dried to obtain Nostoc commune protein peptide;

[0050] S5. Maillard reaction: 15 g of fermented traditional Chinese medicine sugar and 10 g of Nostoc commune protein peptide are mixed evenly, added with 30 g of water, transferred to a hydrothermal reaction kettle, heated to 130 °C, and stirred for 60 min to obtain a Maillard reaction product;

[0051] S6. Metal ion chelation: 10 g of the Maillard reaction product is added to 200 mL of water, 2 g of calcium nitrate and 1 g of magnesium nitrate are added, stirred for 40 min, and dried to obtain a Maillard reaction product chelated with Ca / Mg;

[0052] S7. Preparation of the fermented whitening drink with Taizui probiotics: 12 g of the fermented extract, 10 g of the selenium-enriched Taizui fermented extract, 7 g of the Maillard reaction product chelated with Ca / Mg, and 3 g of trehalose are mixed and added to 150 g of purified water, pasteurized, and filled to obtain the fermented whitening drink with Taizui probiotics.

[0053] Example 3

[0054] This example provides a method for preparing a fermented whitening drink with Taizui probiotics, which includes the following steps:

[0055] S1. Activation of fermenting bacteria: Lactobacillus plantarum, Lactobacillus rhamnosus and selenium-enriched yeast are respectively inoculated into Gause's medium, cultured at 30 °C and 150 r / min for 18 - 24 h to obtain a bacterial strain seed solution with a bacterial content of 10 8 -10 9 cfu / mL;

[0056] S2. Fermentation of traditional Chinese medicine: 11 g of wolfberry, 4 g of houttuynia cordata, 3 g of angelica dahurica and 2 g of Chinese angelica are washed, mixed, crushed, added to 500 mL of water, sterilized, and inoculated with Lactobacillus plantarum and Lactobacillus rhamnosus bacterial strain seed solutions, with inoculation amounts of 1.5 v / v% and 2.5 v / v% respectively. Under anaerobic conditions, cultured at 37 °C and 150 r / min for 42 h, filtered, ethanol is added to the filtrate until the ethanol content in the system is 75 wt%, precipitated for 1.5 h, the solid is washed and dried to obtain traditional Chinese medicine fermented sugar; the ethanol in the filtrate is recovered and dried to obtain a fermented extract;

[0057] S3. Fermentation of Taizui: 10 g of Taizui is washed, minced, added to 200 mL of water, and inoculated with a selenium-enriched yeast bacterial strain seed solution with an inoculation amount of 1.5 v / v%. Cultured at 27 °C and 150 r / min for 30 h, filtered, and the filtrate is freeze-dried to obtain a selenium-enriched Taizui fermented extract;

[0058] S4. Extraction of Nostoc commune protein: Nostoc commune algal dry powder is added to water, with a solid-liquid ratio of 1:8 g / mL, cooled with liquid nitrogen, thawed at room temperature, heated to 50 °C with 1500 W microwave for 3 h, filtered, ethanol is added to the filtrate until the ethanol content in the system is 75 wt%, precipitated for 1 h, filtered, the filtrate is collected, the ethanol in the filtrate is recovered and freeze-dried to obtain Nostoc commune protein peptide;

[0059] S5. Maillard reaction: 13 g of traditional Chinese medicine fermented sugar and 8 g of Nostoc commune protein peptide are mixed evenly, 25 g of water is added, transferred to a hydrothermal reaction kettle, heated to 125 °C, and stirred and reacted for 50 min to obtain a Maillard reaction product;

[0060] S6. Metal ion chelation: 10 g of the Maillard reaction product is added to 200 mL of water, 1.5 g of calcium chloride and 0.7 g of magnesium chloride are added, stirred and reacted for 30 min, and dried to obtain a Maillard reaction product chelated with Ca / Mg;

[0061] Preparation of the probiotic fermented whitening drink from T. sinensis: Mix 11 g of fermented extract, 9 g of fermented extract from T. sinensis rich in selenium, 5.5 g of Maillard reaction product chelated with Ca / Mg, and 2.5 g of trehalose, add them to 135 g of purified water, perform pasteurization, and then fill into containers to obtain the probiotic fermented whitening drink from T. sinensis.

[0062] Comparative Example 1

[0063] Compared with Example 3, the difference lies in that the seed solution of Lactobacillus plantarum was not inoculated in step S2.

[0064] Specifically as follows:

[0065] S2. Fermentation of traditional Chinese medicine: Wash 11 g of Chinese wolfberry, 4 g of houttuynia, 3 g of angelica dahurica, and 2 g of Chinese angelica, mix them, crush them, add them to 500 mL of water, sterilize, inoculate the seed solution of Lactobacillus rhamnosus, with an inoculation amount of 4 v / v%, under anaerobic conditions, at 37 °C, 150 r / min, culture for 42 h, filter, add ethanol to the filtrate until the ethanol content in the system is 75 wt%, precipitate for 1.5 h, wash the solid, and dry to obtain the fermented sugar of traditional Chinese medicine; recover ethanol from the filtrate and dry to obtain the fermented extract.

[0066] Comparative Example 2

[0067] Compared with Example 3, the difference lies in that the seed solution of Lactobacillus rhamnosus was not inoculated in step S2.

[0068] Specifically as follows:

[0069] S2. Fermentation of traditional Chinese medicine: Wash 11 g of Chinese wolfberry, 4 g of houttuynia, 3 g of angelica dahurica, and 2 g of Chinese angelica, mix them, crush them, add them to 500 mL of water, sterilize, inoculate the seed solution of Lactobacillus plantarum, with an inoculation amount of 4 v / v%, under anaerobic conditions, at 37 °C, 150 r / min, culture for 42 h, filter, add ethanol to the filtrate until the ethanol content in the system is 75 wt%, precipitate for 1.5 h, wash the solid, and dry to obtain the fermented sugar of traditional Chinese medicine; recover ethanol from the filtrate and dry to obtain the fermented extract.

[0070] Comparative Example 3

[0071] Compared with Example 3, the difference lies in that calcium chloride was not added in step S6.

[0072] Specifically as follows:

[0073] S6. Metal ion chelation: Add 10 g of Maillard reaction product to 200 mL of water, add 2.2 g of magnesium chloride, stir and react for 30 min, and dry to obtain the Maillard reaction product chelated with Mg.

[0074] Comparative Example 4

[0075] Compared with Example 3, the difference lies in that magnesium chloride is not added in step S6.

[0076] Specifically as follows:

[0077] S6. Metal ion chelation: Add 10 g of the Maillard reaction product to 200 mL of water, add 2.2 g of calcium chloride, stir and react for 30 min, and dry to obtain the Maillard reaction product chelated with Ca.

[0078] Comparative Example 5

[0079] Compared with Example 3, the difference lies in that step S6 is not carried out.

[0080] Specifically as follows:

[0081] S1. Activation of fermentation bacteria: Inoculate Lactobacillus plantarum, Lactobacillus rhamnosus, and selenium-enriched yeast into Gao's medium respectively, and culture at 30 °C and 150 r / min for 18 - 24 h to obtain a strain seed solution with a bacterial content of 10 8 -10 9 cfu / mL;

[0082] S2. Fermentation of traditional Chinese medicine: Wash 11 g of wolfberry, 4 g of houttuynia cordata, 3 g of angelica dahurica, and 2 g of Chinese angelica, mix, crush, add to 500 mL of water, sterilize, inoculate the Lactobacillus plantarum and Lactobacillus rhamnosus strain seed solutions, with inoculation amounts of 1.5 v / v% and 2.5 v / v% respectively. Under anaerobic conditions, culture at 37 °C and 150 r / min for 42 h, filter, add ethanol to the filtrate until the ethanol content in the system is 75 wt%, precipitate for 1.5 h, wash the solid, and dry to obtain the fermented sugar of traditional Chinese medicine; Recover ethanol from the filtrate, dry to obtain the fermented extract;

[0083] S3. Fermentation of Ganoderma lucidum: Wash 10 g of Ganoderma lucidum, crush it, add to 200 mL of water, add the selenium-enriched yeast strain seed solution, with an inoculation amount of 1.5 v / v%, culture at 27 °C and 150 r / min for 30 h, filter, and freeze-dry the filtrate to obtain the selenium-enriched Ganoderma lucidum fermented extract;

[0084] S4. Extraction of Nostoc commune protein: Add Nostoc commune algae dry powder to water, with a solid-liquid ratio of 1:8 g / mL, cool with liquid nitrogen, thaw at room temperature, heat with 1500 W microwave to 50 °C, extract for 3 h, filter, add ethanol to the filtrate until the ethanol content in the system is 75 wt%, precipitate for 1 h, filter, collect the filtrate, recover ethanol from the filtrate, and freeze-dry to obtain Nostoc commune protein peptide;

[0085] S5. Maillard reaction: Mix 13 g of the fermented sugar of traditional Chinese medicine and 8 g of Nostoc commune protein peptide evenly, add 25 g of water, transfer to a hydrothermal reaction kettle, heat to 125 °C, stir and react for 50 min to obtain the Maillard reaction product;

[0086] Preparation of the fermented whitening drink with probiotics from Ganoderma lucidum: Mix 11 g of fermented extract, 9 g of fermented extract from Ganoderma lucidum rich in selenium, 5.5 g of Maillard reaction product, and 2.5 g of trehalose, add them to 135 g of purified water, perform pasteurization, and fill to obtain the fermented whitening drink with probiotics from Ganoderma lucidum.

[0087] Comparative Example 6

[0088] Compared with Example 3, the difference lies in that no fermented extract was added in step S7.

[0089] Specifically as follows:

[0090] Preparation of the fermented whitening drink with probiotics from Ganoderma lucidum: Mix 9 g of fermented extract from Ganoderma lucidum rich in selenium, 5.5 g of Maillard reaction product chelated with Ca / Mg, and 2.5 g of trehalose, add them to 135 g of purified water, perform pasteurization, and fill to obtain the fermented whitening drink with probiotics from Ganoderma lucidum.

[0091] Comparative Example 7

[0092] Compared with Example 3, the difference lies in that no fermented extract from Ganoderma lucidum rich in selenium was added in step S7.

[0093] Specifically as follows:

[0094] Preparation of the fermented whitening drink with probiotics from Ganoderma lucidum: Mix 11 g of fermented extract, 5.5 g of Maillard reaction product chelated with Ca / Mg, and 2.5 g of trehalose, add them to 135 g of purified water, perform pasteurization, and fill to obtain the fermented whitening drink with probiotics from Ganoderma lucidum.

[0095] Comparative Example 8

[0096] Compared with Example 3, the difference lies in that no Maillard reaction product chelated with Ca / Mg was added in step S7.

[0097] Specifically as follows:

[0098] Preparation of the fermented whitening drink with probiotics from Ganoderma lucidum: Mix 11 g of fermented extract, 9 g of fermented extract from Ganoderma lucidum rich in selenium, and 2.5 g of trehalose, add them to 135 g of purified water, perform pasteurization, and fill to obtain the fermented whitening drink with probiotics from Ganoderma lucidum.

[0099] Test Example 1 Antioxidant Activity Test

[0100] 1. DPPH Radical Scavenging Rate

[0101] Take 1 mL of the beverage solution prepared in Examples 1-3 or Comparative Examples 1-8, add 1 mL of 0.2 mmol / L DPPH solution, mix well, react in the dark at room temperature for 30 min, and then measure the absorbance of the sample at a wavelength of 517 nm. At the same time, the blank group is a mixture of 1 mL of the sample solution and 1 mL of methanol solution, and the control group is a mixture of 1 mL of DPPH solution and 1 mL of methanol. Each concentration is repeated 3 times and the average value is taken. Use 0.1 g / L vitamin C solution as the control. The calculation formula is as follows:

[0102] DPPH radical scavenging rate (%) = [1 - (A 样品 - A 空白 ) / A 对照 × 100%

[0103] 2. ABTS radical scavenging rate

[0104] Take 0.15 mL of the beverage solution prepared in Examples 1-3 or Comparative Examples 1-8 respectively, add 2.85 mL of ABTS radical solution, mix well, let stand at room temperature for 10 min, and then measure the absorbance of the sample at a wavelength of 734 nm. At the same time, conduct a blank group experiment (replace the sample solution with 0.15 mL of distilled water). Each concentration is repeated 3 times and the average value is taken. Use 0.1 g / L vitamin C solution as the control. The calculation formula is as follows:

[0105] ABTS radical scavenging rate (%) = [1 - A 样品 / A 空白 × 100%

[0106] The results are shown in Table 1.

[0107] Table 1

[0108] Group DPPH Free Radical Scavenging Rate (%) ABTS Free Radical Scavenging Rate (%) Vitamin C 56.37±6.89 66.12±7.21 Example 1 92.66±10.25 97.89±9.25 Example 2 92.75±9.19 97.53±10.14 Example 3 93.10±9.73 98.05±10.05 Comparative Example 1 87.56±10.56 91.25±9.28 Comparative Example 2 86.68±9.49 89.58±10.55 Comparative Example 3 90.55±10.28 92.39±9.68 Comparative Example 4 90.12±10.05 93.18±10.03 Comparative Example 5 88.69±9.82 90.04±9.82 Comparative Example 6 89.04±9.11 91.12±11.94 Comparative Example 7 85.66±11.56 87.21±10.47 Comparative Example 8 82.01±10.14 84.45±9.94

[0109] As can be seen from the above table, the Taizui probiotic fermented whitening beverage prepared in Examples 1-3 of the present invention has good antioxidant performance.

[0110] Test Example 2 Tyrosinase inhibition activity test

[0111] A 96-well plate was used for sample loading, and 0.2 mol / L PBS buffer was used as the solvent. Tyrosinase was configured to 100 U / mL, and L-DOPA was configured to 0.5 mg / mL. The experimental groups were as follows: blank group, containing 200 μL PBS; blank control group, containing 100 μL PBS + 100 μL tyrosinase solution; sample control group, containing 100 μL PBS + 100 μL beverage solution prepared in Example 1-3 or Comparative Example 1-8; test sample group, containing 100 μL tyrosinase solution + 100 μL beverage solution prepared in Example 1-3 or Comparative Example 1-8; positive control group was arbutin, containing 100 μL tyrosinase solution + 100 μL 3 mg / mL arbutin solution. The sample was incubated at room temperature for 30 minutes, 100 μL of L-DOPA was added, and the incubation continued for 5 minutes. The absorbance (A) value (475 nm) was measured in an ELISA instrument. The experiment was repeated 3 times, and the inhibition rate (R) of tyrosinase was calculated according to formula (2). The results are shown in Table 2.

[0112]

[0113] Among them, A A : Blank A value; A B : Blank control A value; A C : Test sample control A value; A D : Test sample A value.

[0114] Table 2

[0115] Group Tyrosinase Inhibition Rate (%) Arbutin 52.18±5.78 Example 1 74.56±8.93 Example 2 75.09±8.31 Example 3 75.27±7.89 Comparative Example 1 66.57±9.25 Comparative Example 2 64.26±8.59 Comparative Example 3 68.67±9.41 Comparative Example 4 67.92±8.05 Comparative Example 5 65.26±7.96 Comparative Example 6 63.92±9.44 Comparative Example 7 60.11±8.67 Comparative Example 8 58.78±9.91

[0116] It can be seen from the above table that the Tai Sui probiotic fermented whitening beverages prepared in Examples 1-3 of the present invention have good tyrosinase inhibition activity.

[0117] Test Example 3 Melanin content inhibition experiment

[0118] B16F10 melanoma cells were seeded in 6-well plates (2×10 5 / well), add 2mL / well of the fermented whitening beverage made by the probiotics of Example 1-3 or Comparative Example 1-8 on the second day. After different test substances act on the cells for 1h in advance, add 1μmol / L of the inducer α-melanocyte stimulating hormone (α-MSH), and set up a control group (containing only α-MSH). After the two act together for 48h, the cells are collected for subsequent determination, and the final results are calibrated with cell counts. Collect the cell precipitate in each well and add 0.4mL 1mol / L NaOH solution (containing 10% DMSO by volume), incubate at 80°C for 30min, and detect the absorbance at 490nm. Take arbutin 0.1μmol / L as the positive control group. Each group was tested 3 times and the average value was taken. The results are shown in Table 3.

[0119] Inhibitory rate of melanin content (%) = (A control group - A test group) / A control group × 100%

[0120] Table 3

[0121] Group Melanin Content Inhibition Rate (%) Arbutin 4.66±1.13 Example 1 16.74±3.29 Example 2 16.89±2.68 Example 3 17.12±3.05 Comparative Example 1 13.62±4.66 Comparative Example 2 13.19±4.01 Comparative Example 3 14.57±4.29 Comparative Example 4 14.82±3.84 Comparative Example 5 13.95±3.95 Comparative Example 6 12.17±3.67 Comparative Example 7 10.75±3.82 Comparative Example 8 9.24±3.54

[0122] As can be seen from the above table, the Taizui probiotic fermented whitening drink prepared in Examples 1-3 of the present invention has a good effect on inhibiting melanin production.

[0123] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a probiotic fermented whitening beverage, characterized in that: The Chinese medicine fermented sugar and fermented extract are obtained by separating wolfberry, houttuynia cordata, angelica dahurica and angelica sinensis through lactic acid fermentation, and the protein peptides obtained by extracting kudzu vine and the Chinese medicine fermented sugar undergo Maillard reaction, and then calcium and magnesium ions are chelated to obtain the Maillard reaction product of chelated Ca / Mg, which is mixed with selenium-rich Tai Sui fermented extract, fermented extract and trehalose, added into purified water, sterilized, and filled to obtain Tai Sui probiotic fermented whitening beverage; The preparation method comprises the following steps: S1. Activation of fermentation bacteria: Lactobacillus plantarum, Lactobacillus rhamnosus and selenium-enriched yeast were inoculated into Gao's medium respectively, activated and cultured to obtain bacterial seed liquid; S2. Fermentation of Chinese medicine: Wash wolfberry, houttuynia cordata, angelica dahurica and angelica sinensis, mix, crush, add to water, sterilize, inoculate Lactobacillus plantarum and Lactobacillus rhamnosus seed liquid, ferment and culture, filter, add ethanol to the filtrate for precipitation, wash the solid, dry, and obtain Chinese medicine fermentation sugar; recover ethanol from the filtrate, dry, and obtain a fermentation extract; S3. Tai Sui fermentation: Wash and mince Tai Sui, add to water, add selenium-enriched yeast seed solution, ferment, filter, freeze-dry the filtrate to obtain selenium-enriched Tai Sui fermentation extract; S4. Extraction of Gexianmi protein: The Gexianmi algae powder was added to water, cooled with liquid nitrogen, melted at room temperature, extracted by microwave heating, filtered, the filtrate was added with ethanol for precipitation, filtered, the filtrate was recovered with ethanol, freeze-dried, and the protein peptide of Gexianmi was obtained; S5. Maillard reaction: The fermented Chinese medicine sugar and the protein peptide of Puerariae lobata were mixed evenly, water was added, and the mixture was transferred to a hydrothermal reactor, and heated and stirred to react to obtain a Maillard reaction product; S6. Metal ion chelation: adding the Maillard reaction product to water, adding calcium salt and magnesium salt, stirring to react, and drying to obtain a Maillard reaction product chelated with Ca / Mg; S7. Preparation of Tai Sui probiotic fermented whitening drink: The fermented extract, selenium-enriched Tai Sui fermented extract, Maillard reaction product of chelated Ca / Mg and trehalose are mixed and added into purified water, sterilized and filled to prepare Tai Sui probiotic fermented whitening drink.

2. The preparation method according to claim 1, characterized in that: The activation culture conditions in step S1 are 25-35°C, 100-200r / min, and culture for 18-24h. The bacterial content of the bacterial seed liquid is 10 8 -10 9 cfu / mL.

3. The preparation method according to claim 1, characterized in that: The mass ratio of wolfberry, houttuynia cordata, angelica dahurica and angelica sinensis in step S2 is 10-12:3-5:2-4:1-3, the inoculation amounts of the plant lactobacillus and rhamnosus lactobacillus species seed solutions are 1-2v / v% and 2-3v / v%, respectively, the fermentation and culture conditions are anaerobic conditions, 35-38°C, 100-200r / min, culture for 36-48h, ethanol is added until the ethanol content of the system is 70-80wt%, and precipitation is 1-2h.

4. The preparation method according to claim 1, characterized in that: The inoculation amount of the selenium-enriched yeast strain seed liquid in step S3 is 1-2 v / v%, and the fermentation culture conditions are 25-30° C., 100-200 r / min, and the culture is 24-36 h.

5. The preparation method according to claim 1, characterized in that: In step S4, the solid-liquid ratio of the dried powder of Artemisia geum to water is 1:7-10 g / mL, the power of the microwave heating extraction is 1000-2000 W, the temperature is 45-55° C., the time is 2-4 h, ethanol is added until the ethanol content of the system is 70-80 wt%, and the precipitation is 0.5-1 h.

6. The preparation method according to claim 1, characterized in that: In step S5, the mass ratio of the Chinese medicine fermented sugar, the kudzu vine protein peptide and water is 12-15:7-10:20-30, the temperature of the heating and stirring reaction is 120-130° C., and the time is 40-60 min.

7. The preparation method according to claim 1, characterized in that: In step S6, the mass ratio of the Maillard reaction product, the calcium salt and the magnesium salt is 10:1-2:0.5-1, the calcium salt is calcium chloride or calcium nitrate, the magnesium salt is magnesium chloride or magnesium nitrate, and the stirring reaction time is 20-40 min.

8. The preparation method according to claim 1, characterized in that: The mass ratio of the fermentation extract, selenium-enriched Tai Sui fermentation extract, Maillard reaction product of chelated Ca / Mg, trehalose and purified water in step S7 is 10-12:8-10:4-7:2-3:120-150.

9. A whitening beverage fermented by probiotics of Tai Sui prepared by the preparation method according to any one of claims 1 to 8.

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