Fermented mulberry powder with sleep-aiding and sugar-resisting effects as well as preparation method and application of fermented mulberry powder

Abstract

The invention relates to the technical field of mulberry processing, in particular to fermented mulberry powder with sleep-aiding and sugar-resisting effects as well as a preparation method and application of the fermented mulberry powder. The preparation method comprises the following steps: adding mulberries into water, and pulping to prepare pulp; adding a compound enzyme into the slurry for enzymolysis to obtain an enzymatic hydrolysate; equally dividing the enzyme solution into two parts, and respectively adding saccharomyces cerevisiae and plant lactobacillus for first-stage fermentation to obtain a first fermentation solution; sterilizing the first fermentation liquor, and adding casei paracasei for second-stage fermentation to obtain second fermentation liquor; and filtering the second fermentation liquor, adding a solid filler into the filtrate, and then sterilizing and drying in sequence to obtain the fermented mulberry powder. Compared with common mulberry powder, the fermented mulberry powder prepared by the preparation method disclosed by the invention has the effects of aiding sleep and resisting sugar, is obvious in aiding sleep, can inhibit generation of AGEs, and has the effect of inhibiting glycosylation.

Description

Technical Field

[0001] The invention relates to the technical field of mulberry processing, and in particular to fermented mulberry powder with sleep-aiding and anti-sugar effects, and a preparation method and application thereof. Background Art

[0002] Mulberry is also known as mulberry, mulberry seed, mulberry fruit, black mulberry, etc. In ancient my country, mulberry has always been used as a medicinal material. The Compendium of Materia Medica records that "taking the juice after mashing it can relieve alcohol poisoning" and the Southern Yunnan Materia Medica records that "it enters the liver and kidney meridians, benefits the joints of the five internal organs, promotes blood circulation, and long-term consumption can make hair darker and eyes brighter." Modern scientific research shows that mulberry has many physiological effects, which is related to the rich polysaccharides and polyphenols in mulberry, such as anthocyanins, flavanones and phenolic acids. Therefore, in 2002, my country included mulberry in the list of items that are both food and medicine in the relevant notice on food raw material management issued by the Ministry of Health and Law

[2002] No. 51.

[0003] Anthocyanins belong to the flavonoids and are a type of water-soluble natural pigment that is widely found in nature. They are widely found in blue-purple fruits and vegetables. Anthocyanins have excellent nutritional and medicinal value and can also be used as natural edible pigments with high development value. The anthocyanins in mulberry mainly include cyanidin-3-O-glucoside (C3G), cyanidin-3-rutinoside and cyanidin.

[0004] Melatonin (MT) is a substance that can regulate sleep. Its physiological effects include adjusting the biological clock, inducing sleep, delaying aging, regulating the endocrine system, etc. Foods fermented by microorganisms contain not only melatonin, but also a large amount of melatonin isomers.

[0005] Glycation refers to a slow, non-enzymatic reaction between the amino group of a protein and the carbonyl group of a reducing sugar. The reaction first forms an unstable and reversible schif base, which then undergoes molecular rearrangement to eventually form a complex, stable covalent polymer with diverse structures and irreversibility, called advanced glycation end products (AGEs for short). Excessive generation and accumulation of AGEs can cause irreversible damage to the body. Therefore, many scholars are studying the screening of AGE inhibitors from plant foods and Chinese herbal medicines that are both medicinal and edible. Studies have shown that C3G reduces the formation of AGEs by inhibiting key enzymes in the glycation reaction, such as glycosyltransferases, thereby inhibiting the glycation reaction. In addition, C3G can also scavenge free radicals and reduce oxidative stress, thereby protecting cells from damage by the glycation reaction.

[0006] Therefore, fermented mulberry products have become a hot research direction in recent years. Common ones include mulberry wine, mulberry vinegar, and mulberry enzymes, most of which are liquid dosage forms. As the shelf life of liquid dosage forms increases, the content of functional nutrients tends to decay faster. The functional nutrients in solid (powder) dosage forms decay much more slowly during the shelf life than in liquid dosage forms. However, the processing of liquid fermented mulberry products to solid (powder) mulberry products will inevitably lose some functional nutrients. If organic solvents are used for extraction, or more expensive equipment (such as vacuum freeze-drying equipment) is used, it is contrary to food regulations or has high costs, which is often not conducive to industrialization. Therefore, the development of a general process for fermented mulberry powder rich in functional nutrients is one of the technical foundations for improving the level of mulberry industrialization.

[0007] For example, the "Effect of Different Drying Methods on the Content of Active Ingredients in Mulberry" published by Huang Xiaodan et al. shows that different drying methods have great differences in the effects on the quality of mulberry dry matter and the content of active ingredients. The retention rate of active ingredients such as anthocyanins, total flavonoids, total sugars, and reducing sugars in mulberry dried fruits obtained by vacuum freeze-drying is the highest, with the lowest loss and the best fruit shape retention; while the loss rate of hot air drying and far-infrared drying is higher, which has a greater impact on the overall quality of mulberry dried fruits.

[0008] Therefore, how to prepare a fermented mulberry powder rich in functional nutrients is a technical problem that technical personnel in this field urgently need to solve. Summary of the invention

[0009] In order to solve the problem of how to prepare a fermented mulberry powder rich in functional nutrients, the present invention provides a method for preparing fermented mulberry powder with sleep-aiding and anti-sugar effects. The technical scheme is as follows, comprising the following steps: Pre-treatment: Mix mulberries with water to make pulp; Enzymatic hydrolysis: mixing the slurry with a composite enzyme, performing enzymatic hydrolysis, and obtaining an enzymatic hydrolyzate; wherein the composite enzyme comprises pectinase and acid protease; First stage fermentation: taking part of the enzymatic hydrolysate, adding saccharomyces cerevisiae to carry out the first stage A fermentation; taking another part of the enzymatic hydrolysate, adding Lactobacillus plantarum to carry out the first stage B fermentation, and obtaining the first fermentation liquid A liquid and B liquid; Second stage fermentation: the first fermentation liquid A and liquid B are mixed and sterilized, and Lactobacillus paracasei is added to carry out second stage fermentation to obtain a second fermentation liquid; Liquid treatment: filtering the second fermentation liquid, adding a solid filler to the filtrate, and then sterilizing and drying in sequence to obtain the fermented mulberry powder with sleep-inducing and anti-sugar effects; Wherein, the Lactobacillus plantarum is Lactobacillus plantarum YYS-99, and its preservation number is CGMCC No.25838; the Lactobacillus paracasei is Lactobacillus paracasei YYS-K1, and its preservation number is CGMCC No.26405.

[0010] Furthermore, in the pretreatment step, the mulberries are cleaned and mixed with water, and a pulping machine is used to make a slurry; the mass ratio of the mulberries to water is (1-2): (0.25-4).

[0011] Furthermore, in the enzymolysis step, pectinase and acid protease are dissolved in water, mixed with the slurry, heated to 45-60°C for enzymolysis for 1-5h, and then cooled to 25-30°C to obtain an enzymolysis solution; The mass ratio of the pectinase, acid protease, water and mulberry raw material is (0.0002-0.0032): (0.0001-0.0016): (0.002-0.064): (1-2).

[0012] Furthermore, in the first fermentation step, brewer's yeast is mixed with water at 20 to 30° C. and then slowly stirred and mixed, and then part of the enzymatic hydrolyzate is added for fermentation treatment, the fermentation temperature is 20 to 30° C., and the fermentation time is 24 to 72 hours, so as to obtain the first fermentation liquid A; Take another portion of the enzymatic hydrolysate, mix Lactobacillus plantarum YYS-99 with water at 34-40° C. and add the mixture thereto, and perform fermentation at a temperature of 34-40° C. for 24-72 hours to obtain a first fermentation liquid B; Wherein, the mass ratio of the brewer's yeast, water and mulberry raw material is (0.0005-0.012): (0.01-0.24): (1-2); The mass ratio of the plant lactobacillus YYS-99, water and mulberry raw material is (0.0005-0.01): (0.01-0.2): (1-2).

[0013] Furthermore, in the second fermentation step, the sterilization conditions of the first fermentation liquid A and liquid B are sterilized at 70-90° C. for 10-40 min, and then cooled to 26-32° C. to obtain a first fermentation liquid sterilization liquid; The Lactobacillus paracasei YYS-K1 was mixed with water at 26-32° C., and then added into the sterilized solution of the first fermentation solution and mixed, and the fermentation temperature was controlled at 26-32° C. and the fermentation time was 96-144 hours to obtain the second fermentation solution; The mass ratio of the Lactobacillus paracasei YYS-K1, water and mulberry raw material is (0.0005-0.012): (0.01-0.24): (1-2).

[0014] Furthermore, in the feed liquid treatment step, the second fermentation liquid is heated to 70-90° C. and maintained for 10-40 minutes to terminate the fermentation, and then filtered to obtain a clarified filtrate, and the solid filler is fully dissolved in the filtrate so that the solid mass content of the solution is 15%-35%, and then sterilized and dried in sequence to obtain the fermented mulberry powder with sleep-aiding and anti-sugar effects; The mass ratio of the solid filler to the mulberry raw material is (0.05-0.4): (1-2).

[0015] Preferably, the solid content of the solution is adjusted to 15% to 35% by adding water or without adding water.

[0016] Furthermore, in the step of treating the liquid, the liquid is treated by ultra-high temperature instantaneous sterilization at 105-160° C. for 10-40 seconds; and the solid filler is maltodextrin.

[0017] Furthermore, in the step of treating the liquid feed, spray drying is used for drying; The spray drying conditions are: inlet air temperature 135-195°C, feed screw pump motor 5-15Hz, atomization motor 30-50Hz, outlet air temperature 75-105°C.

[0018] The present invention also provides a fermented mulberry powder, which is prepared by the method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects as described above.

[0019] The present invention also provides an application of the fermented mulberry powder as described above in functional food, nutritional food or health food.

[0020] Based on the above, compared with the prior art, the method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects of the present invention has the following beneficial effects: 1. Mulberry is a raw material that is both medicinal and edible, and is rich in functional ingredients. The preparation method provided by the present invention uses specific strains for two-stage temperature-controlled fermentation during the deep processing process, which greatly retains the effective ingredients of mulberry and is easy to industrialize; 2. Compared with common mulberry powder or common fermented mulberry powder, the retention rates of the two functional nutrients MT and C3G in the fermented mulberry powder prepared by the preparation method provided by the present invention are significantly improved; 3. The fermented mulberry powder prepared by the preparation method provided by the present invention can significantly regulate sleep, significantly inhibit the formation of AGEs, and has the effect of inhibiting glycation. DETAILED DESCRIPTION

[0021] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. 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.

[0022] The present invention provides an operational example of a method for preparing fermented mulberry powder, and the specific steps are as follows: Step 1. Pre-treatment: The mulberries are cleaned and mixed with water, and a pulping machine is used to make a slurry; wherein the mass ratio of the mulberry raw material to water is (1-2): (0.25-4).

[0023] Step 2. Enzymatic hydrolysis: Dissolve pectinase and acid protease in water, mix with the slurry, heat to 45-60°C for enzymolysis for 1-5 hours, then cool to 25-30°C to obtain enzymolysis solution; The mass ratio of the pectinase, acid protease, water and mulberry raw material is (0.0002-0.0032): (0.0001-0.0016): (0.002-0.064): (1-2).

[0024] Step 3. First stage fermentation: The enzymatic hydrolysate is evenly divided into 2 portions; the brewer's yeast is mixed with 20-30°C water and then slowly stirred and mixed, and then added into 1 portion of the enzymatic hydrolysate, and the fermentation temperature is controlled at 20-30°C and the fermentation time is controlled at 24-72h to perform fermentation treatment, so as to obtain the first fermentation liquid A; Mix Lactobacillus plantarum YYS-99 with water at 34-40° C., and then add it into another portion of the enzymatic hydrolyzate and mix it evenly. Fermentation is performed at a temperature of 34-40° C. and a fermentation time of 24-72 hours to obtain a first fermentation liquid B. Wherein, the mass ratio of the brewer's yeast, water and mulberry raw material is (0.0005-0.012): (0.01-0.24): (1-2); The mass ratio of the plant lactobacillus YYS-99, water and mulberry raw material is (0.0005-0.01): (0.01-0.2): (1-2).

[0025] Step 4. Secondary fermentation The first fermentation liquid A and liquid B are mixed and sterilized, and the sterilization conditions are as follows: sterilize at 70-90° C. for 10-40 minutes, and then cool to 26-32° C. to obtain the first fermentation liquid sterilization liquid; Lactobacillus paracasei YYS-K1 is mixed with 26-32° C. water, and then added to the first fermentation liquid sterilization liquid and mixed, and the fermentation temperature is controlled at 26-32° C. and the fermentation time is 96-144 hours to obtain the second fermentation liquid; The mass ratio of the Lactobacillus paracasei YYS-K1, water and mulberry raw material is (0.0005-0.012): (0.01-0.24): (1-2).

[0026] Step 5. Liquid treatment: The second fermentation liquid is heated to 70-90° C. and maintained for 10-40 minutes to terminate the fermentation, and then the clarified filtrate is filtered out; the solid filler is fully dissolved in the filtrate, and the solid mass content of the solution is adjusted to 15%-35% by adding water or not adding water, and then sterilized and dried in sequence to obtain the fermented mulberry powder with sleep-aiding and anti-sugar effects; The mass ratio of the solid filler to the mulberry raw material is (0.05-0.4): (1-2); the solid filler is maltodextrin; The sterilization is specifically carried out by ultra-high temperature instantaneous sterilization at 105-160°C for 10-40s; The drying is specifically carried out by spray drying; the spray drying conditions are: inlet air temperature 135-195°C, feed screw pump motor 5-15Hz, atomization motor 30-50Hz, outlet air temperature 75-105°C.

[0027] The present invention also provides the following examples and comparative examples to verify the effects of the solutions of the present invention: Example 1 1. Raw material pretreatment: 60 kg of mulberries were cleaned, mixed with 48 kg of pure water, and made into pulp by a pulping machine, and then added into a temperature-controlled fermentation tank; 2. Enzymatic hydrolysis: 0.018 kg of pectinase and 0.03 kg of acid protease were dissolved in 0.96 kg of pure water and mixed with mulberry pulp. The mixture was heated to 55°C and enzymolyzed for 2 hours. The mixture was then cooled to 25°C to obtain an enzymolysis solution.

[0028] 3. First stage fermentation: Divide the above enzymatic hydrolysate into 2 equal parts; 0.11 kg of brewer's yeast was mixed with 2.18 kg of 30°C pure water and stirred slowly for 30 minutes, and then one portion of the enzymatic hydrolysate was added and stirred evenly, and the fermentation temperature was controlled at 25±3°C and the fermentation time was 48 hours to obtain the first fermentation liquid A; 0.11 kg of Lactobacillus plantarum YYS-99 (preservation number CGMCC No. 25838) was mixed with 2.18 kg of 37°C pure water, and then another portion of enzymatic hydrolysate was added and stirred evenly. The fermentation temperature was controlled at 37±3°C and the fermentation time was 48 hours to obtain the first fermentation liquid B.

[0029] 4. Second stage fermentation: After mixing the first fermentation liquid A and liquid B, heat them to 75°C, keep them for 20 minutes, then cool them to 29°C, mix 0.22 kg of Lactobacillus paracasei YYS-K1 (preservation number CGMCC No.26405) with 4.4 kg of 29°C pure water, put them into a temperature-controlled fermentation tank and stir them evenly, control the fermentation temperature to 29±3°C, and ferment for 96 hours to obtain the second fermentation liquid.

[0030] 5. Liquid treatment: The fermentation liquid was heated to 85°C and maintained for 15 minutes to terminate the fermentation, and then filtered through a 1000-mesh filter cloth using a filter press to obtain 73.5 kg of a clarified filtrate with a solid content of 12.05%. 13.3 kg of maltodextrin was fully dissolved in the filtrate, and then ultra-high temperature instantaneous sterilization was performed at 135°C for 20 seconds. The powder was dried by centrifugal spray drying equipment, and the spray drying conditions were as follows: liquid temperature 70±5°C, air inlet temperature 160±5°C, feed screw pump motor 5Hz, atomization motor 33Hz, and air outlet temperature 77±2°C. 18.2 kg of fermented mulberry powder was obtained by spray drying.

[0031] Example 2 1. Raw material pretreatment: 80 kg of mulberries were cleaned, mixed with 56 kg of pure water, and made into pulp by a pulping machine, and then added into a temperature-controlled fermentation tank; 2. Enzymatic hydrolysis: Then, 0.024 kg of pectinase and 0.04 kg of acid protease were dissolved in 1.28 kg of pure water and mixed with mulberry pulp. The mixture was heated to 55°C and enzymolyzed for 2 hours. The mixture was then cooled to 25°C to obtain an enzymolyzate.

[0032] 3. First stage fermentation: Divide the above enzymatic hydrolysate into 2 equal parts; 0.14 kg of brewer's yeast was mixed with 2.75 kg of 30°C pure water and stirred slowly for 30 minutes, and then one portion of the enzymatic hydrolysate was added and stirred evenly, and the fermentation temperature was controlled at 25±3°C and the fermentation time was 48 hours to obtain the first fermentation liquid A; 0.14 kg of Lactobacillus plantarum YYS-99 (preservation number CGMCC No. 25838) was mixed with 2.75 kg of 37°C pure water, and then another portion of enzymatic hydrolysate was added and stirred evenly. The fermentation temperature was controlled at 37±3°C and the fermentation time was 48 hours to obtain the first fermentation liquid B.

[0033] 4. Second stage fermentation: After mixing the first fermentation liquid A and liquid B, heat them to 75°C, keep them for 20 minutes, then cool them to 29°C, mix 0.28 kg of Lactobacillus paracasei YYS-K1 (preservation number CGMCC No.26405) with 5.5 kg of 29°C pure water, put them into a temperature-controlled fermentation tank and stir them evenly, control the fermentation temperature to 29±3°C, and ferment for 96 hours to obtain the second fermentation liquid.

[0034] 5. Liquid treatment: The fermentation liquid was heated to 85°C and maintained for 15 minutes to terminate the fermentation, and then filtered through a 1000-mesh filter cloth using a filter press to obtain 100.6 kg of a clarified filtrate with a solid content of 11.43%. 17.3 kg of maltodextrin was fully dissolved in the filtrate, and then ultra-high temperature instantaneous sterilization was performed at 135°C for 20 seconds. The centrifugal spray drying equipment was used for drying and powdering. The spray drying conditions were as follows: liquid temperature 70±5°C, air inlet temperature 160±5°C, feed screw pump motor 5Hz, atomization motor 33Hz, and air outlet temperature 77±2°C. 25.5 kg of fermented mulberry powder was obtained by spray drying.

[0035] Example 3 1. Raw material pretreatment: 150 kg of mulberries were cleaned, mixed with 120 kg of pure water, and made into pulp by a pulping machine, and then added into a temperature-controlled fermentation tank; 2. Enzymatic hydrolysis: 0.045 kg of pectinase and 0.075 kg of acid protease were dissolved in 2.4 kg of pure water and mixed with mulberry pulp. The mixture was heated to 55°C and enzymolyzed for 2 hours. The mixture was then cooled to 25°C to obtain an enzymolysis solution.

[0036] 3. First stage fermentation: Divide the above enzymatic hydrolysate into 2 equal parts; 0.27 kg of brewer's yeast was mixed with 5.45 kg of 30°C pure water and stirred slowly for 30 minutes, and then one portion of the enzymatic hydrolysate was added and stirred evenly, and the fermentation temperature was controlled at 25±3°C and the fermentation time was 48 hours to obtain the first fermentation liquid A; 0.27 kg of Lactobacillus plantarum YYS-99 (preservation number CGMCC No. 25838) was mixed with 5.45 kg of 37°C pure water, and then another portion of enzymatic hydrolysate was added and stirred evenly. The fermentation temperature was controlled at 37±3°C and the fermentation time was 48 hours to obtain the first fermentation liquid B.

[0037] 4. Second stage fermentation: After mixing the first fermentation liquid A and liquid B, heat them to 75°C, keep them for 20 minutes, then cool them to 29°C, mix 0.55 kg of Lactobacillus paracasei YYS-K1 (preservation number CGMCC No.26405) with 11 kg of 29°C pure water, put them into a temperature-controlled fermentation tank and stir them evenly, control the fermentation temperature to 29±3°C, and ferment for 96 hours to obtain the second fermentation liquid.

[0038] 5. Liquid treatment: The fermentation liquid was heated to 85°C and maintained for 15 minutes to terminate the fermentation, and then filtered through a 1000-mesh filter cloth using a filter press to obtain 200.8 kg of a clarified filtrate with a solid content of 11.5%, and 34.6 kg of maltodextrin was fully dissolved in the filtrate, and then ultra-high temperature instantaneous sterilization was performed at 135°C for 20 seconds; The centrifugal spray drying equipment was used for drying and powdering. The spray drying conditions were as follows: liquid temperature 70±5°C, air inlet temperature 160±5°C, feed screw pump motor 5Hz, atomization motor 33Hz, and air outlet temperature 77±2°C. 25.5 kg of fermented mulberry powder was obtained by spray drying.

[0039] Comparative Example 1: Ordinary mulberry powder, no fermentation at all Compared with Example 1, the only difference is that this comparative example does not undergo fermentation treatment, that is, the liquid treatment and spray drying are directly performed in sequence after enzymatic hydrolysis, and the other steps and process conditions are consistent with those of Example 1.

[0040] Comparative Example 2: Ordinary mulberry powder, only the first fermentation Compared with Example 1, the only difference is that this comparative example does not undergo a two-stage fermentation treatment, that is, after enzymatic hydrolysis, a first-stage fermentation, a liquid treatment and a spray drying are sequentially performed, and the other steps and process conditions are consistent with those of Example 1.

[0041] Comparative Example 3: Ordinary mulberry powder, only the first stage A fermentation Compared with Example 1, the only difference is that this comparative example does not carry out the first stage B fermentation and the second stage fermentation treatment, that is, after enzymatic hydrolysis, the first stage A fermentation, liquid treatment and spray drying are carried out in sequence, and the other steps and process conditions are consistent with Example 1.

[0042] Comparative Example 4: Ordinary mulberry powder, only the first stage B fermentation Compared with Example 1, the only difference is that this comparative example does not carry out the first stage A fermentation and the second stage fermentation treatment, that is, after enzymatic hydrolysis, the first stage B fermentation, liquid treatment and spray drying are carried out in sequence, and the other steps and process conditions are consistent with Example 1.

[0043] Comparative Example 5: Ordinary mulberry powder, only the second fermentation Compared with Example 1, the only difference is that the comparative example does not undergo the first fermentation treatment, that is, the second fermentation, liquid treatment and spray drying are directly carried out in sequence after enzymatic hydrolysis, and the other steps and process conditions are consistent with those of Example 1.

[0044] Comparative Example 6: Ordinary mulberry powder, fermented in non-specific order Compared with Example 1, the only difference is that the first fermentation and the second fermentation steps of this comparative example are reversed, that is, the fermentation treatment with Lactobacillus paracasei YYS-K1 is carried out first, and then the fermentation treatment with Saccharomyces cerevisiae and Lactobacillus plantarum YYS-99 is carried out. The other steps and process conditions are consistent with those of Example 1.

[0045] Comparative Example 7: Ordinary mulberry powder, fermented under non-specific fermentation conditions The specific conditions and process are as follows: 60 kg of mulberries are cleaned, mixed with 48 kg of pure water through a pulping machine to make pulp, and then added to a temperature-controlled fermentation tank; 0.018 kg of pectinase and 0.03 kg of acid protease are dissolved in 0.96 kg of pure water and mixed with the mulberry pulp, then heated to 55°C, enzymolyzed for 2 hours, and then cooled to 43°C; Divide the above enzymatic hydrolysate into 2 equal parts; 0.11 kg of brewer's yeast was mixed with 2.18 kg of 35°C pure water and stirred slowly for 30 minutes. Then one portion of the enzymatic hydrolysate was added and stirred evenly. The fermentation temperature was controlled at 35±3°C and the fermentation time was 48 hours. 0.11 kg of Lactobacillus plantarum YYS-99 (CGMCC No. 25838) was mixed with 2.18 kg of 45°C pure water, and then another portion of the enzymatic hydrolysate was added and stirred evenly. The fermentation temperature was controlled at 45±3°C and the fermentation time was 48 hours. Heat a section of fermentation liquid to 75°C, maintain for 20 minutes, then cool to 40°C, mix 0.22 kg of Lactobacillus paracasei YYS-K1 (CGMCC No. 26405) with 4.4 kg of 40°C pure water, put it into the temperature-controlled fermentation tank and stir well, control the fermentation temperature at 40±3°C, and ferment for 96 hours.

[0046] Other steps and process conditions are consistent with those in Example 1 Comparative Example 8: Ordinary mulberry powder, fermented with non-specific strains Compared with Example 1, the only difference is: The other commercially available Lactobacillus plantarum types were used to replace Lactobacillus plantarum YYS-99, and the other commercially available Lactobacillus paracasei types were used to replace Lactobacillus paracasei YYS-K1. Other steps and process conditions were the same as those in Example 1.

[0047] The raw material strains used in the above examples and comparative examples have been published and collected: Among them, Lactobacillus plantarum (Lactiplantibacillus plantarum )YYS-99, Lactobacillus paracasei ( Lacticaseibacillus paracasei ) YYS-K1 is disclosed in the Chinese invention patent applications with publication numbers CN115960767A and CN116478863A, respectively. The technicians can obtain it from the deposit center according to the deposit number. The deposit information is as follows: Lactobacillus plantarum YYS-99, Latin name: Lactiplantibacillus plantarum , deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, the deposit date is September 28, 2022, and the deposit number is CGMCC No.25838.

[0048] Lactobacillus paracasei YYS-K1, classified and named as Lactobacillus paracasei YYS-K1, Latin name: Lacticaseibacillusparacasei, deposited in the General Microbiology Center of China Microorganism Culture Collection Administration Committee, deposit number: CGMCC No.26405. Deposit address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, deposit date: January 6, 2023. Note: This strain is also known as Lactobacillus paracasei YYS-K1. Due to the replacement of the old and new catalogs of strain classification names, two different names appear, and the two refer to the same essence.

[0049] Performance Testing (1) Determination of melatonin content The determination of melatonin (MT) content refers to the first method in "GBT5009.170-2003 Determination of Melatonin Content in Health Foods", that is, the MT content of the comparative example and the experimental group (ie, the embodiment) was determined by high performance liquid chromatography-ultraviolet detection method, as shown in Table 1: Table 1 MT content of mulberry powder Group MT content (ng / g) Remark Comparative Example 1 2.197 No fermentation at all Comparative Example 2 7.251 First fermentation only Comparative Example 3 5.989 Only the first stage A fermentation Comparative Example 4 3.528 Only the first stage B fermentation Comparative Example 5 3.817 Second fermentation only Comparative Example 6 6.215 Non-specific order fermentation Comparative Example 7 3.229 Non-specific fermentation conditions Comparative Example 8 3.055 Non-specific bacterial fermentation Experimental Group 8.537 Example 1 Experimental Group 8.269 Example 2 Experimental Group 8.926 Example 3 The content unit ng / g represents the MT content in 1g of mulberry powder. The results show that the MT content of the mulberry powder in the experimental group of the embodiment is significantly higher than the MT content of the comparative example (ordinary mulberry powder) ( P <0.05).

[0050] (2) Determination of cyanidin-3-O-glucoside content The determination of cyanidin-3-O-glucoside (C3G) content was carried out with reference to Zhang Tingting's "One-test-multiple-evaluation method for determination of the contents of three anthocyanins in Aronia nigra fruit", and the C3G content of the comparative example and the experimental group (ie, the embodiment) was determined by HPLC, as shown in Table 2: Table 2 C3G content of mulberry powder Group C3G content (mg / g) Remark Comparative Example 1 0.125 No fermentation at all Comparative Example 2 0.184 First fermentation only Comparative Example 3 0.130 Only the first stage A fermentation Comparative Example 4 0.191 Only the first stage B fermentation Comparative Example 5 0.207 Second fermentation only Comparative Example 6 0.337 Non-specific order fermentation Comparative Example 7 0.134 Non-specific fermentation conditions Comparative Example 8 0.158 Non-specific bacterial fermentation Experimental Group 0.524 Example 1 Experimental Group 0.473 Example 2 Experimental Group 0.540 Example 3 The content unit mg / g represents the C3G content in 1g of mulberry powder. The results show that the C3G content of the mulberry powder in the experimental group of the embodiment is significantly higher than the C3G content of the comparative example (ordinary mulberry powder) ( P <0.05).

[0051] (3) Tests to inhibit AGEs production Taking the positive drug aminoguanidine as the positive control, the test results of the inhibition rate of fluorescent AGEs by the fermented mulberry powder prepared in the above Examples 1-3 and Comparative Examples 1-8 are shown in Table 3 below, wherein the specific test method for the inhibition rate of fluorescent AGEs refers to Zhou Huiji's "Multi-model Evaluation of the Anti-glycation Effect of Sophora japonica Water Extract and Study on Its Active Ingredients", the positive drug aminoguanidine and fermented mulberry powder were prepared into samples of different concentrations using solvent water, and their inhibition rates on fluorescent AGEs were tested.

[0052] Table 3 Inhibition rate of fluorescent AGEs in Examples and Comparative Examples

[0053] As can be seen from Table 3, compared with the control example, the inhibition rate of the fermented mulberry powder prepared in Examples 1-3 on fluorescent AGEs is significantly improved; compared with the positive drug aminoguanidine group, as the concentration gradually increases, the inhibition rate of the fermented mulberry powder on fluorescent AGEs is also significantly improved; in addition, fermented mulberry powder of different concentrations has a certain in vitro activity of inhibiting the formation of AGEs, and shows an obvious dose-dependent relationship.

[0054] The present invention also provides an application case of fermented mulberry powder Application example 1: Fermented mulberry solid beverage (1) The specific formula of fermented mulberry powder for preparing solid beverages is shown in Table 4 below: Table 4 Fermented mulberry solid beverage formula

[0055] (2) Specific operation steps: 1. Weighing: Weigh the raw materials respectively according to the above formula ratio.

[0056] 2. Premixing: Premix ingredients 4 (vitamin C) to 7 (silicon dioxide) until well combined.

[0057] 3. Big Mix: All materials were mixed in the mixer for 25 min, and the semi-finished product was checked for uniformity and fluidity upon discharge.

[0058] 4. Inner packaging: The evenly mixed solid beverage is packaged into semi-finished products with a specification of 8g / bag.

[0059] 5. Outer packaging: The solid beverage semi-finished product with inner packaging is packed into boxes to obtain the fermented mulberry solid beverage finished product.

[0060] Application Example 2: Fermented mulberry tablet candy (1) The specific formula of fermented mulberry powder used in compressed candy is shown in Table 5 below: Table 5 Fermented mulberry tablet candy formula

[0061] (2) Specific operation steps: 1. Weighing: Weigh the raw materials respectively according to the above formula ratio.

[0062] 2. Premixing: Premix ingredients 3 (microcrystalline cellulose) to 7 (vitamin C) until well combined.

[0063] 3. Big Mix: All materials were mixed in the mixer for 25 min, and the semi-finished product was checked for uniformity and fluidity upon discharge.

[0064] 4. Tableting: The mixed powder is directly compressed into 1.0 g / tablet candies using a tablet press.

[0065] 5. Inner packaging: The evenly mixed solid beverage is packaged into semi-finished compressed candies in the specification of 1 piece / bag.

[0066] 6. Outer packaging: The semi-finished compressed candy with inner packaging is packed into boxes to obtain the finished fermented mulberry compressed candy.

[0067] Application Example 3: Fermented Mulberry Beverage (1) The specific formula of fermented mulberry powder used in preparing beverages is shown in Table 6 below: Table 6 Fermented mulberry beverage recipe

[0068] (2) Specific operation steps: 1. Weighing: Weigh the raw materials respectively according to the above formula ratio.

[0069] 2. Dissolution: Add raw materials 2 (fermented mulberry powder) to raw material 5 (potassium sorbate) to raw material 1 (pure water) in sequence, and stir until completely dissolved.

[0070] 3. Cut: Heat the solution in step 2 to 70-75°C, pour it into a shear tank, add raw material 6 (pectin), and shear for 13 minutes.

[0071] 4. Add flavor: When step 3 is completed, immediately add material 7 (compound berry essence) and continue shearing for 2 minutes.

[0072] 5. Sterilization: The material solution in step 4 was sterilized by ultra-high temperature instantaneous sterilization at a temperature of 138°C for 20 seconds.

[0073] 6. Filling: The sterilized liquid in step 5 is filled into semi-finished products in a sterile workshop.

[0074] 7. Outer packaging: The filled semi-finished product is packed into boxes to obtain the finished product of fermented mulberry beverage.

[0075] In summary, the method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects provided by the present invention has at least the following technical effects: 1. Mulberry is a raw material that is both medicinal and edible, and is rich in functional ingredients. The preparation method provided by the present invention uses specific strains for two-stage temperature-controlled fermentation during the deep processing process, which greatly retains the effective ingredients of mulberry and is easy to industrialize; 2. Compared with common mulberry powder or common fermented mulberry powder, the retention rates of the two functional nutrients MT and C3G in the fermented mulberry powder prepared by the preparation method provided by the present invention are significantly improved; 3. The fermented mulberry powder prepared by the preparation method provided by the present invention can significantly regulate sleep, significantly inhibit the formation of AGEs, and has the effect of inhibiting glycation.

[0076] It should be noted that: (1) Definition: In this article, “~” is used to indicate a numerical range, and the range indicated by this expression includes two endpoint values.

[0077] The term "food" as used herein is broad in nature and includes food and drink for humans, and the food products are suitable for and designed for human consumption. The above application examples are only examples of the fermented mulberry powder prepared by the present invention for use in solid beverages, compressed candies, beverages and other foods. It can also be used to prepare other solid preparations such as powders, tablets, gels, etc., and dispersed in liquids to prepare liquid preparations, including but not limited to the application example solutions. Similarly, the auxiliary materials for preparing various dosage forms of food can also be selected from existing commercially available auxiliary materials, including but not limited to the application example solutions.

[0078] Ultra-high temperature instantaneous sterilization is a sterilization process, abbreviated as UHT (Ultra-high temperature instantaneous sterilization).

[0079] In summary, the specific parameters or some commonly used reagents or raw materials in the above embodiments are specific embodiments or preferred embodiments under the concept of the present invention, but are not intended to limit the present invention; those skilled in the art can make adaptive adjustments within the concept and protection scope of the present invention.

[0080] In addition, unless otherwise specified, the raw materials used may also be conventional commercial products in the art, or prepared by conventional methods in the art; that is, the reagents and instruments used in this embodiment are not indicated with information such as the manufacturer, and are all conventional products that can be purchased on the market.

[0081] Although the terms such as enzymolysis, sterilization, one-stage fermentation and two-stage fermentation are used more frequently in this article, the possibility of using other terms is not excluded. These terms are used only to more conveniently describe and explain the essence of the present invention; interpreting them as any additional limitation is contrary to the spirit of the present invention.

[0082] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing fermented mulberry powder with sleep-aiding and anti-sugar effects, characterized in that: The following steps are involved: Pre-treatment: Mix mulberries with water to make pulp; Enzymatic hydrolysis: mixing the slurry with a composite enzyme, performing enzymatic hydrolysis, and obtaining an enzymatic hydrolyzate; wherein the composite enzyme comprises pectinase and acid protease; First stage fermentation: taking part of the enzymatic hydrolysate, adding saccharomyces cerevisiae to carry out the first stage A fermentation; taking another part of the enzymatic hydrolysate, adding Lactobacillus plantarum to carry out the first stage B fermentation, and obtaining the first fermentation liquid A liquid and B liquid; Second stage fermentation: the first fermentation liquid A and liquid B are mixed and sterilized, and Lactobacillus paracasei is added to carry out second stage fermentation to obtain a second fermentation liquid; Liquid treatment: filtering the second fermentation liquid, adding a solid filler to the filtrate, and then sterilizing and drying in sequence to obtain the fermented mulberry powder with sleep-inducing and anti-sugar effects; Wherein, the Lactobacillus plantarum is Lactobacillus plantarum YYS-99, and its preservation number is CGMCC No.25838; the Lactobacillus paracasei is Lactobacillus paracasei YYS-K1, and its preservation number is CGMCC No.26405.

2. The method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects according to claim 1, characterized in that: In the pretreatment step, the mulberries are cleaned and mixed with water, and a pulping machine is used to make a slurry; the mass ratio of the mulberries to the water is (1-2): (0.25-4).

3. The method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects according to claim 1, characterized in that: In the enzymatic hydrolysis step, pectinase and acid protease are dissolved in water, mixed with the slurry, heated to 45-60°C for enzymatic hydrolysis for 1-5 hours, and then cooled to 25-30°C to obtain an enzymatic hydrolyzate; The mass ratio of the pectinase, acid protease, water and mulberry raw material is (0.0002-0.0032): (0.0001-0.0016): (0.002-0.064): (1-2).

4. The method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects according to claim 1, characterized in that: In the first fermentation step, brewer's yeast is mixed with water at 20-30° C. and then slowly stirred and mixed, and then part of the enzymatic hydrolyzate is added for fermentation treatment at a fermentation temperature of 20-30° C. for a fermentation time of 24-72 hours to obtain a first fermentation liquid A; Take another portion of the enzymatic solution, mix Lactobacillus plantarum YYS-99 with 34-40℃ water and add it. Fermentation is performed at a temperature of 34 to 40° C. for 24 to 72 hours to obtain a first fermentation liquid B; Wherein, the mass ratio of the brewer's yeast, water and mulberry raw material is (0.0005-0.012): (0.01-0.24): (1-2); The mass ratio of the plant lactobacillus YYS-99, water and mulberry raw material is (0.0005-0.01): (0.01-0.2): (1-2).

5. The method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects according to claim 1, characterized in that: In the second fermentation step, the first fermentation liquid A and liquid B are sterilized at 70-90°C for 10-40 minutes, and then cooled to 26-32°C to obtain a first fermentation liquid sterilization liquid; The Lactobacillus paracasei YYS-K1 was mixed with water at 26-32° C., and then added into the sterilized solution of the first fermentation solution and mixed, and the fermentation temperature was controlled at 26-32° C. and the fermentation time was 96-144 hours to obtain the second fermentation solution; The mass ratio of the Lactobacillus paracasei YYS-K1, water and mulberry raw material is (0.0005-0.012): (0.01-0.24): (1-2).

6. The method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects according to claim 1, characterized in that: In the liquid processing step, the second fermentation liquid is heated to 70-90° C. and maintained for 10-40 minutes to terminate the fermentation, and then filtered to obtain a clarified filtrate, and the solid filler is fully dissolved in the filtrate so that the solid content of the solution is 15%-35%, and then sterilized and dried in sequence to obtain the fermented mulberry powder with sleep-aiding and anti-sugar effects; The mass ratio of the solid filler to the mulberry raw material is (0.05-0.4): (1-2).

7. The method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects according to claim 1, characterized in that: In the liquid processing step, the liquid is processed by ultra-high temperature instantaneous sterilization at 105-160° C. for 10-40 seconds; and the solid filler is maltodextrin.

8. The method for preparing the fermented mulberry powder with sleep-aiding and anti-sugar effects according to claim 1, characterized in that: In the liquid processing step, spray drying is used for drying; The spray drying conditions are: inlet air temperature 135-195°C, feed screw pump motor 5-15Hz, atomization motor 30-50Hz, outlet air temperature 75-105°C.

9. A fermented mulberry powder, characterized in that: The fermented mulberry powder with sleep-aiding and anti-sugar effects is prepared by the preparation method of any one of claims 1 to 8.

10. Use of the fermented mulberry powder according to claim 9 in functional food, nutritional food or health food.

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