Fermented whitening beverage containing ganoderma lucidum probiotics and preparation method of fermented whitening beverage

Abstract

The invention provides a ganoderma lucidum probiotic-containing fermented whitening beverage and a preparation method thereof, and belongs to the technical field of beverages. Chinese wolfberry fruits, heartleaf houttuynia herbs, radix angelicae and Chinese angelica are subjected to lactic acid bacteria fermentation separation to obtain traditional Chinese medicine fermented sugar and a fermentation extract, protein peptide extracted from nostoc sphaeroids kutz and the traditional Chinese medicine fermented sugar are subjected to a Maillard reaction, calcium and magnesium ions are chelated, and a Ca / Mg chelated Maillard reaction product is obtained; and mixing with selenium-rich ganoderma lucidum fermentation extract, the fermentation extract and trehalose, adding into purified water, sterilizing, and filling to obtain the ganoderma lucidum probiotic fermented whitening beverage. The ganoderma lucidum probiotic fermented whitening beverage prepared by the preparation method is a compound beverage, has relatively good antioxidant, anti-inflammatory, whitening and anti-aging effects, is relatively good in taste, relatively long in shelf life and convenient to store, is deeply loved by consumers, and has a wide application prospect.

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 invention provides a method for preparing a Tai Sui probiotic fermented whitening beverage. The method comprises the following steps: subjecting wolfberry, houttuynia cordata, angelica dahurica and angelica sinensis to lactic acid fermentation separation to obtain Chinese medicinal fermented sugar and a fermented extract; after protein peptides obtained by extracting kudzu vine and the Chinese medicinal fermented sugar undergo Maillard reaction, calcium and magnesium ions are chelated to obtain a Maillard reaction product of chelated Ca / Mg; the product is mixed with a selenium-rich Tai Sui fermented extract, a fermented extract and trehalose, and added into purified water; the product is sterilized and filled to obtain a Tai Sui probiotic fermented whitening beverage.

[0008] As a further improvement of the present invention, the following steps are included:

[0009] S1. Activation of fermentation bacteria: Lactobacillus plantarum, Lactobacillus rhamnosus and selenium-enriched yeast were inoculated into Gao's medium respectively, and activated and cultured to obtain bacterial seed liquid;

[0010] S2. Fermentation of Chinese medicine: Wash wolfberry, houttuynia, angelica and angelica, mix, crush, add to water, sterilize, inoculate Lactobacillus plantarum and Lactobacillus rhamnosus seed solution, 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;

[0011] S3. Tai Sui fermentation: Wash Tai Sui, mince, add to water, add selenium-enriched yeast strain seed solution, fermentation culture, filter, freeze-dry the filtrate to obtain selenium-enriched Tai Sui fermentation extract;

[0012] S4. Extraction of Kudzu protein: The Kudzu dry 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, and freeze-dried to obtain Kudzu protein peptide;

[0013] S5. Maillard reaction: The fermented Chinese medicine sugar and the kudzu vine protein peptide are mixed evenly, water is added, and the mixture is transferred into a hydrothermal reactor, heated and stirred to react to obtain a Maillard reaction product;

[0014] S6. Metal ion chelation: adding the Maillard reaction product to water, adding calcium salt and magnesium salt, stirring for reaction, and drying to obtain a Maillard reaction product chelated with Ca / Mg;

[0015] S7. Preparation of Tai Sui probiotic fermented whitening drink: 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.

[0016] As a further improvement of the present invention, 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 -109 cfu / mL.

[0017] As a further improvement of the present invention, 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 seed solutions are 1-2v / v% and 2-3v / v%, respectively, the fermentation and culture conditions are under 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.

[0018] As a further improvement of the present invention, the inoculation amount of the selenium-enriched yeast 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.

[0019] As a further improvement of the present invention, the solid-to-liquid ratio of the dry powder of Semen Armeniacae Sinensis and water in step S4 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, and ethanol is added until the ethanol content of the system is 70-80 wt%, and the precipitation is 0.5-1 h.

[0020] As a further improvement of the present invention, in step S5, the mass ratio of the Chinese medicinal fermentation sugar, the kudzu vine protein peptide and the 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, the mass ratio of the Maillard reaction product, the calcium salt and the magnesium salt in step S6 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, the mass ratio of the fermentation extract, the selenium-enriched Tai Sui fermentation extract, the 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.

[0023] The present invention further protects a whitening beverage fermented by probiotics of Tai Sui 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 and then to dopaquinone by tyrosinase, and melanin is formed by the action of DHI oxidase and DHICA oxidase. Tyrosinase is a key enzyme and rate-limiting enzyme in the synthesis of melanin in organisms. It is a copper-containing oxidoreductase. The presence of oxygen is a necessary condition in its catalytic process of melanin synthesis. Therefore, antioxidants can reduce the synthesis of melanin by blocking or weakening the activity of tyrosinase. Substances that have both antioxidant and tyrosinase inhibitory activity can combine the effects of the two to better reduce melanin synthesis. Secondly, antioxidants can also reduce free radicals produced by various exogenous factors, thereby avoiding the oxidation of collagen fibers in the skin (appearing yellow) and causing yellowish skin color.

[0026] In the present invention, wolfberry has good antioxidant and whitening effects; the representative active ingredient tannic acid substance in Houttuynia cordata extract has whitening, moisturizing, antioxidant, anti-inflammatory and other effects; the volatile oils and coumarins in Angelica dahurica can effectively inhibit the synthesis of intracellular melanin and the activity of tyrosinase; the effective ingredients of Angelica sinensis, volatile oils, phenolic compounds, coumarin compounds, terpenoid compounds, cis-apigenin, etc., all show obvious inhibitory effects on tyrosinase; the four medicines are used together to invigorate the spleen and stomach, regulate the qi, promote blood circulation and whiten the skin, but if the traditional Chinese medicine components are directly decocted, the extraction is incomplete, the process is cumbersome, it is easy to mold after being left for a long time, the volume of the medicine is large, and it is inconvenient to carry. The present invention adopts lactic acid bacteria for fermentation, which not only extracts efficiently and promotes the rupture of plant cell walls, but also produces more new substances with better activity. At the same time, the fermentation products of lactic acid bacteria mainly include organic acids, extracellular polysaccharides, polypeptides and bacteriocins, etc., have antioxidant effects, can eliminate free radicals in cells and reduce the accumulation of reactive oxygen species (ROS) in the body, greatly improve the antioxidant, anti-inflammatory and whitening activities, and the obtained fermentation products are separated to obtain polysaccharides and fermentation extracts, the extraction method is simple, and after drying, it is easy to carry and not easy to deteriorate.

[0027] Gexianmi is a type of algae 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, the cell wall of Gexianmi algae is broken, thereby promoting the dissolution of active components in the algae under microwave heating conditions. After alcohol precipitation, the polysaccharides are separated to obtain Gexianmi algae protein peptides.

[0028] The Maillard reaction refers to a complex chemical reaction between amino groups in amino acids or proteins and carbonyl groups in reducing sugars under heating conditions. In the present invention, the protein peptides of Puerariae lobata and fermented sugars of traditional Chinese medicine are subjected to the Maillard reaction under heating conditions, and the prepared products such as melanoidin have significant antioxidant activity, and other products such as polyphenols, flavonoids and other active ingredients have the effects of nourishing qi and blood, anti-cancer, antibacterial, preventing and treating diseases, and thus have excellent antioxidant, anti-aging and whitening effects.

[0029] Calcium ions are one of the most common metal ions in the skin, and play an important role in skin tissue regeneration, maintaining skin integrity, strengthening skin barriers, promoting epidermal cell proliferation and differentiation, remodeling skin extracellular matrix, as well as whitening, soothing and many other functional areas. Magnesium ions can promote the repair of skin barriers. After the skin is immersed in a salt bath solution rich in magnesium ions, the skin tolerance is improved, the skin barrier function and stratum corneum hydration are improved, and the skin roughness is improved.

[0030] The present invention mixes the prepared Maillard reaction product with calcium salt and magnesium salt for reaction, thereby greatly improving the fixation amount of metal ions, preventing the metal ions from being lost after entering the human body, promoting their absorption in the intestine, and greatly improving the absorption amount of metal ions by the human body, thereby better exerting the effect of whitening and improving the skin.

[0031] The invention ferments the Gastrodia elata with selenium-rich yeast, and the obtained selenium-rich Gastrodia elata fermentation extract is rich in selenium, has good antioxidant effect, and improves the body's immunity. At the same time, the composite bacteria in the Gastrodia elata and its products have good antioxidant and whitening effects, and can also increase the shelf life of the beverage.

[0032] The Tai Sui probiotic fermented whitening beverage prepared by the invention is a composite beverage, has good anti-oxidation, anti-inflammatory, whitening and anti-aging effects, has good taste, has a long shelf life, is easy to store, is deeply loved by consumers, and has broad application prospects. DETAILED DESCRIPTION

[0033] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] Lactobacillus plantarum, 10 billion cfu / g, Lactobacillus rhamnosus, 10 billion cfu / g, were purchased from Zhongke Jiayi Bioengineering Technology Co., Ltd.; Selenium-enriched yeast, 20 billion cfu / g, were purchased from Angel Yeast Co., Ltd. Gexian Mi Algae dry powder, water content <5%, was purchased from Hunan Yan Di Bioengineering Co., Ltd.

[0035] Example 1

[0036] This embodiment provides a method for preparing a probiotic fermented whitening beverage, comprising the following steps:

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

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

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

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

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

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

[0043] S7. Preparation of Tai Sui probiotic fermented whitening drink: 10g fermentation extract, 8g selenium-rich Tai Sui fermented extract, 4g Maillard reaction product of chelated Ca / Mg and 2g trehalose are mixed and added into 120g purified water, pasteurized and filled to prepare Tai Sui probiotic fermented whitening drink.

[0044] Example 2

[0045] This embodiment provides a method for preparing a probiotic fermented whitening beverage, comprising the following steps:

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

[0047] S2. Fermentation of Chinese medicine: 12g of wolfberry, 5g of houttuynia cordata, 4g of angelica dahurica and 3g of angelica were washed, mixed, crushed, added to 500mL of water, sterilized, inoculated with Lactobacillus plantarum and Lactobacillus rhamnosus seed liquid, the inoculation amount was 2v / v% and 3v / v%, respectively, under anaerobic conditions, 38°C, 200r / min, cultured for 48h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 80wt%, precipitated for 2h, the solid was washed, dried, and Chinese medicine fermentation sugar was obtained; ethanol was recovered from the filtrate and dried to obtain a fermentation extract;

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

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

[0050] S5. Maillard reaction: 15 g of Chinese medicine fermented sugar and 10 g of Pueraria lobata protein peptide were mixed evenly, 30 g of water was added, transferred to a hydrothermal reactor, heated to 130 ° C, stirred and reacted for 60 min to obtain a Maillard reaction product;

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

[0052] S7. Preparation of Tai Sui probiotic fermented whitening drink: 12g fermentation extract, 10g selenium-rich Tai Sui fermented extract, 7g Maillard reaction product of chelated Ca / Mg and 3g trehalose are mixed and added into 150g purified water, pasteurized and filled to prepare Tai Sui probiotic fermented whitening drink.

[0053] Example 3

[0054] This embodiment provides a method for preparing a probiotic fermented whitening beverage, comprising the following steps:

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

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

[0057] S3. Tai Sui fermentation: 10 g Tai Sui was washed, minced, added to 200 mL of water, and selenium-enriched yeast seed solution was added, the inoculation amount was 1.5 v / v%, 27 ° C, 150 r / min, cultured for 30 h, filtered, and the filtrate was freeze-dried to obtain a selenium-enriched Tai Sui fermentation extract;

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

[0059] S5. Maillard reaction: 13 g of Chinese medicine fermented sugar and 8 g of Pueraria lobata protein peptide were mixed evenly, 25 g of water was added, transferred to a hydrothermal reactor, heated to 125 ° C, stirred and reacted for 50 min to obtain a Maillard reaction product;

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

[0061] S7. Preparation of Tai Sui probiotic fermented whitening drink: 11g fermentation extract, 9g selenium-rich Tai Sui fermented extract, 5.5g Maillard reaction product of chelated Ca / Mg and 2.5g trehalose are mixed and added into 135g purified water, pasteurized and filled to prepare Tai Sui probiotic fermented whitening drink.

[0062] Comparative Example 1

[0063] Compared with Example 3, the difference is that the Lactobacillus plantarum seed liquid is not inoculated in step S2.

[0064] The details are as follows:

[0065] S2. Fermentation of traditional Chinese medicine: 11g of wolfberry, 4g of houttuynia cordata, 3g of angelica dahurica and 2g of angelica sinensis were washed, mixed, crushed, added into 500mL of water, sterilized, inoculated with Lactobacillus rhamnosus seed solution at an inoculation rate of 4v / v%, cultured under anaerobic conditions at 37°C, 150r / min for 42h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 75wt%, precipitated for 1.5h, the solid was washed, dried, and traditional Chinese medicine fermentation sugar was obtained; ethanol was recovered from the filtrate, dried, and a fermentation extract was obtained.

[0066] Comparative Example 2

[0067] Compared with Example 3, the difference is that the Lactobacillus rhamnosus seed liquid is not inoculated in step S2.

[0068] The details are as follows:

[0069] S2. Fermentation of traditional Chinese medicine: 11g of wolfberry, 4g of houttuynia cordata, 3g of angelica dahurica and 2g of angelica sinensis were washed, mixed, crushed, added into 500mL of water, sterilized, inoculated with Lactobacillus plantarum seed solution, the inoculation amount was 4v / v%, under anaerobic conditions, 37°C, 150r / min, cultured for 42h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 75wt%, precipitated for 1.5h, the solid was washed, dried, and traditional Chinese medicine fermentation sugar was obtained; ethanol was recovered from the filtrate, dried, and a fermentation extract was obtained.

[0070] Comparative Example 3

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

[0072] The details are as follows:

[0073] S6. Metal ion chelation: 10 g of the Maillard reaction product was added to 200 mL of water, and 2.2 g of magnesium chloride was added. The mixture was stirred for 30 min and dried to obtain a Maillard reaction product chelated with Mg.

[0074] Comparative Example 4

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

[0076] The details are as follows:

[0077] S6. Metal ion chelation: 10 g of the Maillard reaction product was added to 200 mL of water, and 2.2 g of calcium chloride was added. The mixture was stirred for reaction for 30 min and dried to obtain a Maillard reaction product chelated with Ca.

[0078] Comparative Example 5

[0079] Compared with the embodiment 3, the difference is that step S6 is not performed.

[0080] The details are as follows:

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

[0082] S2. Fermentation of Chinese medicine: 11g of wolfberry, 4g of houttuynia cordata, 3g of angelica dahurica and 2g of angelica were washed, mixed, crushed, added to 500mL of water, sterilized, inoculated with Lactobacillus plantarum and Lactobacillus rhamnosus seed liquid, the inoculation amounts were 1.5v / v% and 2.5v / v%, respectively, under anaerobic conditions, 37°C, 150r / min, cultured for 42h, filtered, ethanol was added to the filtrate until the ethanol content of the system was 75wt%, precipitated for 1.5h, the solid was washed, dried, and Chinese medicine fermentation sugar was obtained; ethanol was recovered from the filtrate and dried to obtain a fermentation extract;

[0083] S3. Tai Sui fermentation: 10 g Tai Sui was washed, minced, added to 200 mL of water, and selenium-enriched yeast seed solution was added, the inoculation amount was 1.5 v / v%, 27 ° C, 150 r / min, cultured for 30 h, filtered, and the filtrate was freeze-dried to obtain a selenium-enriched Tai Sui fermentation extract;

[0084] S4. Extraction of Kudzu protein: The Kudzu powder was added to water at a solid-liquid ratio of 1:8 g / mL, cooled with liquid nitrogen, melted at room temperature, heated at 1500 W in a microwave oven to 50 ° C, extracted for 3 h, filtered, and the filtrate was added with ethanol to a system ethanol content of 75 wt %, precipitated for 1 h, filtered, and the filtrate was collected. The filtrate was recovered with ethanol and freeze-dried to obtain Kudzu protein peptide;

[0085] S5. Maillard reaction: 13 g of Chinese medicine fermented sugar and 8 g of Pueraria lobata protein peptide were mixed evenly, 25 g of water was added, transferred to a hydrothermal reactor, heated to 125 ° C, stirred and reacted for 50 min to obtain a Maillard reaction product;

[0086] S6. Preparation of Tai Sui probiotic fermented whitening drink: 11g fermentation extract, 9g selenium-rich Tai Sui fermented extract, 5.5g Maillard reaction product and 2.5g trehalose are mixed and added into 135g purified water, pasteurized and filled to prepare Tai Sui probiotic fermented whitening drink.

[0087] Comparative Example 6

[0088] Compared with Example 3, the difference is that no fermentation extract is added in step S7.

[0089] The details are as follows:

[0090] S7. Preparation of Tai Sui probiotic fermented whitening drink: 9 g of selenium-rich Tai Sui fermented extract, 5.5 g of Maillard reaction product of chelated Ca / Mg and 2.5 g of trehalose are mixed and added into 135 g of purified water, pasteurized and filled to prepare Tai Sui probiotic fermented whitening drink.

[0091] Comparative Example 7

[0092] Compared with Example 3, the difference is that no selenium-rich Gastrodia elata fermentation extract is added in step S7.

[0093] The details are as follows:

[0094] S7. Preparation of Tai Sui probiotic fermented whitening drink: 11 g of fermentation extract, 5.5 g of Maillard reaction product of chelated Ca / Mg and 2.5 g of trehalose are mixed and added into 135 g of purified water, pasteurized and filled to prepare Tai Sui probiotic fermented whitening drink.

[0095] Comparative Example 8

[0096] Compared with Example 3, the difference is that the Maillard reaction product of chelated Ca / Mg is not added in step S7.

[0097] The details are as follows:

[0098] S7. Preparation of Tai Sui probiotic fermented whitening drink: 11g fermented extract, 9g selenium-rich Tai Sui fermented extract and 2.5g trehalose are mixed and added into 135g purified water, pasteurized and filled to prepare Tai Sui probiotic fermented whitening drink.

[0099] Test Example 1 Antioxidant Test

[0100] 1. DPPH free radical scavenging rate

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

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

[0103] 2. ABTS free radical scavenging rate

[0104] Take 0.15 mL of each beverage solution prepared in Examples 1-3 or Comparative Examples 1-8, add 2.85 mL of ABTS free radical solution, mix well, let stand at room temperature for 10 minutes, and measure the absorbance of the sample at a wavelength of 734 nm. At the same time, a blank group experiment (using 0.15 mL of distilled water instead of the sample solution) was carried out, and each concentration was repeated 3 times to take the average value. A 0.1 g / L vitamin C solution was used as a control. The calculation formula is as follows:

[0105] ABTS free 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] 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 antioxidant properties.

[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] Melanin content inhibition rate (%) = (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] 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 a good effect of inhibiting the production of melanin.

[0123] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a probiotic fermented whitening beverage, characterized in that: Chinese medicinal fermentation sugar and fermentation extract are obtained by separating wolfberry, houttuynia cordata, angelica dahurica and angelica sinensis through lactic acid fermentation. The protein peptides obtained by extracting kudzu vine and Chinese medicinal fermentation sugar undergo Maillard reaction, and then calcium and magnesium ions are chelated to obtain Maillard reaction products of chelated Ca / Mg. The products are mixed with selenium-rich Tai Sui fermentation extract, fermentation extract and trehalose, added into purified water, sterilized and filled to make Tai Sui probiotic fermentation whitening beverage.

2. The preparation method according to claim 1, characterized in that: The following steps are involved: S1. Activation of fermentation bacteria: Lactobacillus plantarum, Lactobacillus rhamnosus and selenium-enriched yeast were inoculated into Gao's medium respectively, and activated and cultured to obtain bacterial seed liquid; S2. Fermentation of Chinese medicine: Wash wolfberry, houttuynia, angelica and angelica, mix, crush, add to water, sterilize, inoculate Lactobacillus plantarum and Lactobacillus rhamnosus seed solution, 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 Tai Sui, mince, add to water, add selenium-enriched yeast strain seed solution, fermentation culture, filter, freeze-dry the filtrate to obtain selenium-enriched Tai Sui fermentation extract; S4. Extraction of Kudzu protein: The Kudzu dry 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, and freeze-dried to obtain Kudzu protein peptide; S5. Maillard reaction: The fermented Chinese medicine sugar and the kudzu vine protein peptide are mixed evenly, water is added, and the mixture is transferred into a hydrothermal reactor, 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 for reaction, and drying to obtain a Maillard reaction product chelated with Ca / Mg; S7. Preparation of Tai Sui probiotic fermented whitening drink: 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.

3. The preparation method according to claim 2, 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.

4. The preparation method according to claim 2, 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 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.

5. The preparation method according to claim 2, characterized in that: The inoculation amount of the selenium-enriched yeast strain seed solution 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 carried out for 24-36 hours.

6. The preparation method according to claim 2, characterized in that: In step S4, the solid-to-liquid ratio of the dried powder of Herba Lycopodii and 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.

7. The preparation method according to claim 2, 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, and the temperature of the heating and stirring reaction is 120-130° C. and the time is 40-60 min.

8. The preparation method according to claim 2, 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.

9. The preparation method according to claim 2, 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.

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

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