A double-issued food with sleep improvement efficacy and a preparation method and application thereof

Abstract

This invention provides a dual-action food product with sleep-improving effects, its preparation method, and its application, comprising the following steps: (1) mixing peanut sprout powder with water to obtain a mixture; (2) adjusting the pH of the mixture to 6-8, adding an enzyme preparation for enzymatic hydrolysis to obtain an enzymatic hydrolysis solution; (3) adding a carbon source and a nitrogen source, adjusting the pH of the enzymatic hydrolysis solution to 6-7, and performing enzyme-inactivating extraction to obtain an enzyme-inactivating extract; (4) waiting for the temperature of the enzyme-inactivating extract to drop to 30-40℃, adding a fermenting agent for fermentation to obtain a fermentation liquid; (5) centrifuging the fermentation liquid, and vacuum drying the supernatant to obtain a fermented product. The obtained fermented product has an increased β-NMN content and has the effect of improving sleep, and can be applied to sleep-improving products.

Description

Technical Field

[0001] This invention belongs to the field of fermentation technology, specifically relating to a dual-action food that improves sleep, its preparation method, and its application. Background Technology

[0002] Peanut sprouts are the tender seedlings that grow from sprouted peanuts.

[0003] "Double-fermentation" foods refer to foods made by applying fermentation technology to the processing of sprouted foods. Examples include rice sprout fermented beverages, which are made by sprouting rice and then fermenting it. After undergoing both sprouting and fermentation, the raw materials not only have a unique flavor but also gain beneficial substances, thus improving the nutritional value of the food.

[0004] Insomnia, also known as sleep disorder insomnia, is a common health problem that affects the quality and quantity of sleep. Clinical symptoms of insomnia mainly include difficulty falling asleep, frequent awakenings during the night, early awakening with difficulty falling back asleep, and fatigue and poor concentration upon waking. Long-term insomnia can also lead to complications such as memory loss, mood swings, and weakened immunity. Currently, there are no reports on the sleep-improving effects of fermented peanut sprouts. Summary of the Invention

[0005] Therefore, the present invention provides a dual-action food that improves sleep, its preparation method, and its application.

[0006] The technical content of this invention is as follows:

[0007] On the one hand, the present invention provides a dual-action food for improving sleep, the preparation method of which includes the following steps: (1) mixing peanut sprout powder with water to obtain a mixture; (2) adjusting the pH of the mixture to 6-8, adding an enzyme preparation for enzymatic hydrolysis to obtain an enzymatic hydrolysis solution; (3) adding a carbon source and a nitrogen source, and adjusting the pH of the enzymatic hydrolysis solution to 6-7, and performing enzyme inactivation extraction to obtain an enzyme inactivation extract; (4) waiting for the temperature of the enzyme inactivation extract to drop to 30-40℃, adding a fermenting agent for fermentation to obtain a fermentation liquid; (5) centrifuging the fermentation liquid, and vacuum drying the supernatant to obtain a fermented product.

[0008] Preferably, the fermentation agent includes *Lactobacillus plantarum* FTCM001 and *Lactobacillus plantarum* XD087.

[0009] More preferably, the weight ratio of *Lactobacillus plantarum* FTCM001 to *Lactobacillus plantarum* XD087 is (2-3):(1-2).

[0010] More preferably, the viability count of the *Lactobacillus plantarum* FTCM001 is 1 × 10⁻⁶. 11CFU / g or higher; the viability count of the *Lactobacillus plantarum* XD087 is 1×10⁻⁶. 11 CFU / g or higher.

[0011] Preferably, the weight ratio of the fermenting agent to peanut sprout powder is 1:(50-150); more preferably, the weight ratio of the fermenting agent to peanut sprout powder is 1:100.

[0012] Preferably, the weight ratio of peanut sprout powder to water in step (1) is 1:(3-8); more preferably, the weight ratio of peanut sprout powder to water in step (1) is 1:5.

[0013] Preferably, the weight ratio of the enzyme preparation to peanut sprout powder is 1:(25-45); more preferably, the weight ratio of the enzyme preparation to peanut sprout powder is 1:35.

[0014] Preferably, the enzyme preparation in step (2) includes cellulase and bromelain; more preferably, the weight ratio of cellulase to bromelain is (2-5):(1-2); even more preferably, the weight ratio of cellulase to bromelain is 3:1.

[0015] Preferably, the cellulase has an enzyme activity of 10,000-100,000 U / g, and the bromelain has an enzyme activity of 50,000-100,000 U / g.

[0016] Preferably, the carbon source is a common carbon source in the food industry, specifically including one or more of glucose, molasses, and fructooligosaccharides; the nitrogen source is a common nitrogen source in the food industry, specifically including one or more of corn oligopeptides and peptones.

[0017] Preferably, the enzymatic hydrolysis temperature in step (2) is 45-60℃ and the enzymatic hydrolysis time is 45-70min.

[0018] Preferably, the enzyme inactivation extraction time in step (3) is 15-40 min, and the enzyme inactivation extraction temperature is 80-120℃.

[0019] Preferably, the fermentation time in step (4) is 12-20h and the fermentation temperature is 30-40℃.

[0020] Preferably, in step (5), the vacuum degree of vacuum drying is -0.085MPa, the drying temperature is 40-50℃, and the drying time is 6-7h.

[0021] Preferably, the preparation method of peanut sprout powder is also included, with the following steps: (1) Pretreatment: Select high-quality peanut seeds with mature, round, plump, bright color and no damage to the skin, and soak them in 0.3% sodium hypochlorite solution for 30 minutes; (2) Soaking: Add an appropriate amount of water to submerge the peanut seeds and soak them at room temperature of 20-25℃. After soaking, rinse them twice in an appropriate amount of water to remove some impurities and sodium hypochlorite residue; (3) Germination: Germinate the peanut seeds in a constant temperature incubator for 2 days; (4) Germination: After germination, insert the germinated peanut seeds into a perforated seedling tray for germination in the dark. Water the peanuts about 4 times a day. Each time, water the peanut sprouts thoroughly to avoid the seeds generating too much heat and rotting. Germination ends after 5 days. (5) Vacuum freeze-drying: After germination, perform vacuum freeze-drying to obtain peanut sprout powder.

[0022] Preferably, the soaking time in step (2) is 18-24h; the germination temperature in step (3) is 20-30℃; the pre-freezing temperature for vacuum freeze drying in step (5) is -45℃ to -35℃, the pre-freezing time is 6-10h, the drying temperature is 35-45℃, and the drying time is 25-35h.

[0023] On the other hand, the present invention provides a dual-action food product, which is obtained according to the above preparation method.

[0024] In another aspect, the present invention provides the application of the above-mentioned dual-action food in the preparation of sleep-improving and / or anti-aging products.

[0025] Compared with the prior art, the present invention has the following advantages:

[0026] 1. This invention provides a method for preparing a dual-sleep food, specifically peanut sprouts prepared by enzymatic hydrolysis and fermentation. The inventors found that by selecting cellulase and bromelain for enzymatic hydrolysis, and then fermenting with Lactobacillus plantarum FTCM001 and XD087, the fermentation product has a high content of β-nicotinamide mononucleotide (β-NMN) and has a good effect on improving sleep.

[0027] 2. This invention provides a dual-action food product, which, upon testing, was found to have a high content of β-nicotinamide mononucleotide (β-NMN), making it suitable for use in anti-aging products. Furthermore, mouse experiments have shown that it has a significant effect on improving sleep, making it suitable for use in calming and sleep-aiding products. Detailed Implementation

[0028] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0029] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0030] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels. Products from different manufacturers do not have a significant impact on the effectiveness.

[0031] Cellulase and bromelain were purchased from Nanning Dongheng Huadao Biotechnology Co., Ltd., with cellulase having an enzyme activity of 10,000 U / g and bromelain having an enzyme activity of 100,000 U / g. Lactobacillus plantarum and Leuconostoc mesenteroides subsp. mesenteroides were purchased from Jiangsu Xinshenao Biotechnology Co., Ltd. Lactobacillus plantarum Lp3a with a bacterial activity of 500 billion CFU / g and Leuconostoc mesenteroides subsp. mesenteroides LCM020 with a bacterial activity of 500 billion CFU / g.

[0032] Preparation Example 1

[0033] This preparation example provides a method for preparing peanut sprout powder, including the following steps: (1) Pretreatment: Select high-quality peanut seeds (Luhua No. 9) with mature, round, plump, bright color and no broken skin, and soak them in 0.3% sodium hypochlorite solution for 30 min; (2) Soaking: Add an appropriate amount of water to submerge the peanut seeds and soak them at room temperature of 20℃ for 24 h. After soaking, rinse them twice in an appropriate amount of water to remove some impurities and sodium hypochlorite residue; (3) Germination: Germinate the peanut seeds in a constant temperature incubator at 20℃ for 2 days; (4) Germination: After germination, insert the germinated peanut seeds into a seedling tray with holes for germination in the dark. Water the peanuts about 4 times a day. Each time, water the peanut sprouts thoroughly to avoid the seeds generating too much heat and rotting. Germination ends after 5 days; (5) Vacuum freeze drying: After germination, pre-freeze the peanut sprouts at -45℃ for 6 h and dry them at a partition temperature of 35℃ for 35 h to obtain peanut sprout powder.

[0034] Example 1: Activation of the bacterial strain and preparation of bacterial powder

[0035] Lactobacillus plantarum FTCM001, with accession number CGMCC No. 26813, has been disclosed in patent CN118638687A; Lactobacillus plantarum XD087, with accession number CCTCC M 20241729, has been disclosed in patent CN119662456A.

[0036] Preparation of *Lactobacillus plantarum* FTCM001 and *Lactobacillus plantarum* XD087 bacterial powders: 1 mL of glycerol tubes containing either *Lactobacillus plantarum* FTCM001 or *Lactobacillus plantarum* XD087 were inoculated into 100 mL LMR broth and anaerobically cultured at 37°C for 24 h. The inoculum was then transferred to 100 mL LMR broth at a 2% inoculum size and anaerobically cultured at 37°C for another 24 h. This process was repeated once. The bacterial suspension was centrifuged at 8000 rpm for 10 min at 4°C, the supernatant was discarded, and the suspension was resuspended in 0.9% sterile physiological saline (1 / 10 the original bacterial volume). This process was repeated twice to prepare the bacterial suspension. The bacterial suspension was mixed with a lyophilization protectant (9.5% xylooligosaccharide, 9.5% whole milk powder, and 6% monosodium glutamate) at a ratio of 1:3. The mixture was pre-frozen at -80°C for 2.5 h and then dried under vacuum at 4 Pa ​​for 24 h, yielding viable bacterial counts of 5 × 10⁻⁶. 11 Freeze-dried bacterial powder of active Lactobacillus plantarum FTCM001 or Lactobacillus plantarum XD087 at CFU / g.

[0037] MRS liquid culture medium (g / L): beef extract 25, glucose 30, dipotassium hydrogen phosphate 2, diammonium hydrogen citrate 2, sodium acetate 5, magnesium sulfate 3.2, Tween 80 1.

[0038] Example 2

[0039] This embodiment provides a method for preparing a dual-action food product that improves sleep, comprising the following steps: (1) mixing 500 parts of peanut sprout powder (Preparation Example 1) with 2500 parts of water to obtain a mixture; (2) adjusting the pH of the mixture to 7, adding 10.5 parts of cellulase and 3.5 parts of bromelain, and enzymatically hydrolyzing at 50°C for 60 minutes to obtain an enzymatic hydrolysate; (3) adding 16 parts of fructooligosaccharides and 42 parts of corn oligopeptides, adjusting the pH of the enzymatic hydrolysate to 6.5, and enzymatically hydrolyzing at 50°C for 60 minutes to obtain an enzymatic hydrolysate; (3) Extract the enzyme at 80°C for 40 min to obtain an enzyme-inactivated extract; (4) When the temperature of the enzyme-inactivated extract drops to 37°C, add 3.33 parts of Bacillus plantarum FTCM001 powder (Example 1) and 1.67 parts of Bacillus plantarum XD087 powder (Example 1), and ferment for 18 h to obtain a fermentation broth; (5) Centrifuge the fermentation broth, and dry the supernatant at a vacuum of -0.085 MPa, a drying temperature of 50°C, and a drying time of 6 h to obtain the fermented product.

[0040] Example 3

[0041] This embodiment provides a method for preparing a dual-action food with sleep-improving effects, including the following steps: (1) Take 500 parts of peanut sprout powder and mix with 4000 parts of water to obtain a mixture; (2) Adjust the pH of the mixture to 8, add 14.3 parts of cellulase and 5.7 parts of bromelain, and enzymatically hydrolyze at 45°C for 70 min to obtain an enzymatic hydrolysis solution; (3) Add 16 parts of fructooligosaccharide and 42 parts of corn oligopeptide, adjust the pH of the enzymatic hydrolysis solution to 6, and inactivate the enzyme at 120°C for 15 min to obtain an enzyme-inactivated extract; (4) When the temperature of the enzyme-inactivated extract drops to 40°C, add 2.5 parts of *Lactobacillus plantarum* FTCM001 (Example 1) and 0.83 parts of *Lactobacillus plantarum* XD087 (Example 1), and ferment for 20 h to obtain a fermentation broth; (5) Centrifuge the fermentation broth, and dry the supernatant at a vacuum of -0.085 MPa, a drying temperature of 40°C, and a drying time of 7 h to obtain the fermented product.

[0042] Example 4

[0043] This embodiment provides a method for preparing a dual-action food with sleep-improving effects, including the following steps: (1) Mix 500 parts of peanut sprout powder with 1500 parts of water to obtain a mixture; (2) Adjust the pH of the mixture to 6, add 7.3 parts of cellulase and 3.7 parts of bromelain, and enzymatically hydrolyze at 60°C for 45 min to obtain an enzymatic hydrolysis solution; (3) Add 16 parts of fructooligosaccharide and 42 parts of corn oligopeptide, adjust the pH of the enzymatic hydrolysis solution to 7, and extract with enzyme at 100°C for 20 min to obtain an enzyme-inactivated extract; (4) When the temperature of the enzyme-inactivated extract drops to 30°C, add 5 parts of *Lactobacillus plantarum* FTCM001 (Example 1) and 5 parts of *Lactobacillus plantarum* XD087 (Example 1), and ferment for 12 h to obtain a fermentation liquid; (5) Centrifuge the fermentation liquid, and dry the supernatant at a vacuum of -0.085 MPa, a drying temperature of 45°C, and a drying time of 6.5 h to obtain the fermented product.

[0044] Comparative Example 1

[0045] This comparative example provides a method for preparing a dual-action food product that improves sleep. The difference between this comparative example and Example 2 is that no enzyme preparation is added, and no enzymatic hydrolysis process is used. It includes the following steps:

[0046] (1) Take 500 parts of peanut sprout powder (Preparation Example 1) and mix with 2500 parts of water to obtain a mixture; (2) Adjust the pH of the mixture to 7, stir and extract at 50℃ for 60 min to obtain a treatment solution; (3) Add 16 parts of fructooligosaccharide and 42 parts of corn oligopeptide, adjust the pH of the treatment solution to 6.5, and extract at 80℃ for 40 min to obtain an extract; (4) When the temperature of the extract drops to 37℃, add 3.33 parts of *Lactobacillus plantarum* FTCM001 (Example 1) and 1.67 parts of *Lactobacillus plantarum* XD087 (Example 1), ferment for 18 h to obtain a fermentation broth; (5) Centrifuge the fermentation broth, and dry the supernatant at a vacuum of -0.085 MPa, a drying temperature of 50℃, and a drying time of 6 h to obtain the fermented product.

[0047] Comparative Example 2

[0048] This comparative example provides a method for preparing a dual-action food that improves sleep. The difference from Example 2 is that the enzyme preparation in step (2) only includes cellulase. Specifically, (2) the pH of the mixture is adjusted to 7, 14 parts of cellulase are added, and the mixture is enzymatically hydrolyzed at 50°C for 60 minutes to obtain the enzymatic hydrolysate.

[0049] The rest is the same as in Example 2.

[0050] Comparative Example 3

[0051] This comparative example provides a method for preparing a dual-action food that improves sleep. The difference from Example 2 is that the fermenting agent in step (4) only includes Bacillus plantarum FTCM001. Specifically, (4) the temperature of the enzyme extract is lowered to 37°C, 5 parts of Bacillus plantarum FTCM001 (Example 1) are added, and fermentation is carried out for 18 hours to obtain the fermentation liquid.

[0052] The rest is the same as in Example 2.

[0053] Comparative Example 4

[0054] This comparative example provides a method for preparing a dual-action food that improves sleep. The difference from Example 2 is that the fermenting agent in step (4) only includes Bacillus plantarum XD087. Specifically, (4) the temperature of the enzyme extract is lowered to 37°C, 5 parts of Bacillus plantarum XD087 (Example 1) are added, and fermentation is carried out for 18 hours to obtain the fermentation liquid.

[0055] The rest is the same as in Example 2.

[0056] Comparative Example 5

[0057] This comparative example provides a method for preparing a dual-action food that improves sleep. The difference from Example 2 is that in step (4), the fermenting agent is replaced with commercially available Lactobacillus plantarum XD087. Specifically, (4) when the temperature of the enzyme-inactivated extract is lowered to 37°C, 3.33 parts of Lactobacillus plantarum FTCM001 (Example 1) and 1.67 parts of Lactobacillus plantarum Lp3a are added and fermented for 18 hours to obtain the fermentation liquid; wherein the viable count of Lactobacillus plantarum Lp3a is 500 billion CFU / g.

[0058] The rest is the same as in Example 2.

[0059] Comparative Example 6

[0060] This comparative example provides a method for preparing a dual-action food that improves sleep. The difference from Example 2 is that in step (4), the fermenting agent is replaced with Leuconostoc mesenteroides subsp. enterica LCM020 instead of Bacillus plantarum XD087. Specifically, (4) when the temperature of the enzyme-inactivated extract is lowered to 37°C, 3.33 parts of Bacillus plantarum FTCM001 (Example 1) and 1.67 parts of Leuconostoc mesenteroides subsp. enterica LCM020 are added and fermented for 18 hours to obtain the fermentation liquid; wherein the viable count of Leuconostoc mesenteroides subsp. enterica is 500 billion CFU / g.

[0061] The rest is the same as in Example 2.

[0062] Experimental Example 1: Determination of β-NMN Content - Ultra-High Performance Liquid Chromatography

[0063] 1.1 Sample Preparation

[0064] Weigh 2g of the fermentation products from Examples 2-4 and Comparative Examples 1-6 respectively, dissolve them in 50% acetonitrile solution, sonicate for 20min, place at room temperature, dilute to volume in a 50mL volumetric flask, and filter through a 0.22μm organic filter membrane for subsequent analysis.

[0065] 1.2 Determination Method

[0066] Preparation of standards: A certain amount of β-Nicotinamide mononucleotide (purity ≥95.0%, Sigma) was weighed and dissolved in 50% acetonitrile solution to prepare standard solutions with mass concentrations of 10, 50, 100, 200, 300, 500, and 1000 ng / mL. The linear regression equations were: Y = 46.932X - 284.2691, R0 2 =0.999.

[0067] Chromatographic conditions: SHIM-PACK WAX-2 column (4.0×50mm, 5μm); column temperature: 30℃; flow rate: 0.8mL / min; injection volume: 5μL; detection wavelength: 254nm; mobile phase A: 50mmol / L potassium dihydrogen phosphate; mobile phase B: acetonitrile; gradient elution conditions: within 0–10 min, phase A decreased from 98% to 95%, and phase B increased from 2% to 5%; within 10–15 min, phase A decreased from 95% to 80%, and phase B increased from 5% to 20%; within 15–16 min, phase A increased from 80% to 98%, and phase B decreased from 20% to 2%, held for 9 min and then stopped.

[0068] 1.3 Experimental Results

[0069] As shown in Table 1, the β-NMN content in Examples 2-4 was relatively high, with Example 2 showing a β-NMN content as high as 6.95 μg / g. Comparative Examples 1-6 showed a significant decrease compared to Example 2, with Comparative Examples 1-2 showing significantly lower β-NMN content than Example 2, indicating that the enzymatic hydrolysis process and the type of enzyme preparation affect the β-NMN content. The NMN content in Comparative Examples 3-6 was significantly lower than that in Example 2, indicating that the two strains of this invention are superior fermentation strains for the synergistic fermentation of peanut sprouts.

[0070] Table 1. β-NMN content in each group

[0071]

[0072] Note: Compared with Example 2, #P<0.05, ##P<0.01.

[0073] Experiment Example 2: Sleep Improvement Effect - Mouse Experiment

[0074] 2.1 Experimental animals and housing environment: SPF grade Balb-c healthy male mice, weighing 18~20g, were housed in an environment with a room temperature of 25~27℃, relative humidity of 50~70%, and a light / dark cycle of 12h each, with free access to water and food for one week.

[0075] 2.2 Grouping and Administration: After one week of acclimatization, mice were randomly divided into four groups for direct sleep experiments, pentobarbital-induced sleep duration prolongation experiments, pentobarbital sodium subvalence-induced hypnosis experiments, and pentobarbital sodium sleep latency experiments. Each group was further divided into 10 subgroups (n=10 per subgroup): a blank control group (NC) and experimental groups (Examples 2-4 and Comparative Examples 1-6). The experimental groups were administered the corresponding test substance by gavage at a dose of 15 mL / kg body weight, while the blank control group was given an equal volume of pure water, once a day for 30 consecutive days.

[0076] The gavage volume was calculated using the kilogram-weight conversion factor method. Based on the assumption that an adult weighing 60kg ingests 100mL of oral solution per day, the cross-point conversion factor is 9.01. Therefore, the daily gavage dose for mice should be (100mL / 60kg)×9.01=15mL / kg.

[0077] 2.3 Direct Sleep Experiment

[0078] Mice were administered the medication by gavage for 30 consecutive days. After the last gavage, the number of mice in each group who fell asleep within 1 hour was observed. Sleep was defined as the disappearance of the righting reflex; that is, mice that could not right themselves within 1 minute when placed in a supine position were considered to be asleep.

[0079] 2.4 Pentobarbital sodium sleep duration prolongation experiment: 30 minutes after the last administration of sodium pentobarbital, mice were intraperitoneally injected with sodium pentobarbital at a dose of 50 mg / kg (based on mouse body weight, mice fell asleep 100% but the sleep time was short). The sleep duration (time from the disappearance of the righting reflex to the reappearance of the righting reflex) of each group of mice was observed and recorded, and the sleep time between each group was compared.

[0080] 2.5 Subthreshold dose hypnotic experiment of sodium pentobarbital: 30 minutes after the last administration, sodium pentobarbital was injected intraperitoneally at a dose of 30 mg / kg (based on mouse body weight, the dose that prevents 80%–90% of mice from falling asleep). The number of animals that fell asleep within 30 minutes (those whose righting reflex disappeared for more than 1 minute) was recorded.

[0081] 2.6 Sodium barbital sleep latency test: 30 minutes after the last administration, sodium barbital was injected intraperitoneally at a dose of 230 mg / kg (based on mouse body weight, 100% of mice fell asleep). The time from the disappearance of the righting reflex 1 minute after injection to the disappearance of the righting reflex was recorded as the sleep latency. The effect of the test sample on the sleep latency of sodium barbital was observed.

[0082] 2.7 Statistical methods: Quantitative data were expressed as mean ± standard deviation (x ± s), and analysis of variance was used for comparisons among multiple groups; chi-square test was used for categorical data. A p-value < 0.05 was considered statistically significant.

[0083] 2.8 Result Interpretation: If any two of the three tests—the prolonged sodium pentobarbital sleep time test, the subthreshold dose sodium pentobarbital hypnosis test, and the sodium barbital sleep latency test—are positive and there is no direct sleep effect, the test sample can be determined to have the effect of improving sleep function.

[0084] 2.9 Experimental Results

[0085] 2.9.1 Results of the direct sleep experiment

[0086] Thirty days after the intervention, the sleep status of mice in each group was observed within 1 hour after the last gavage. No righting reflex was lost in any group, and the sleep rate of mice in each group remained at 0%, indicating that peanut sprout fermentation has no direct sleep effect on mice.

[0087] 2.9.2 Results of the experiment on prolonging sodium pentobarbital sleep time

[0088] Compared with the blank control group, the sleep time of mice in Examples 2-4 and Comparative Examples 1-6 was significantly prolonged; compared with Example 2, the sleep time of Comparative Examples 1-2 was shortened, indicating that the enzymatic hydrolysis process and the type of enzyme preparation of the present invention affect the effect of the fermentation product on prolonging the sleep of mice. The sleep time of Comparative Examples 3-6 was significantly lower than that of Example 2, indicating that the types of fermentation agents selected in the present invention are a better combination that can prolong the sleep time.

[0089] Table 2. Effects of peanut sprout fermentation on prolonged pentobarbital sodium sleep time in mice.

[0090]

[0091] Note: Compared with the blank control group, +P<0.05, ++P<0.01; compared with Example 2, #P<0.05, ##P<0.01.

[0092] 2.9.3 Results of the subthreshold dose hypnotic experiment using sodium pentobarbital

[0093] The sleep rate of mice in the blank control group was 10%, verifying the rationality of using a dose of 30 mg / kg sodium pentobarbital to study the effect of subthreshold doses on mouse hypnosis. The sleep rate of mice in Examples 2-4 was significantly higher than that in the blank control group, and the sleep rate of mice in Comparative Examples 1-6 was reduced to varying degrees compared with that in Example 2.

[0094] Table 3 Effects of subthreshold doses of sodium pentobarbital on hypnosis in mice

[0095]

[0096] 2.9.4 Results of the sodium barbital sleep latency test

[0097] Compared with the blank control group, Examples 2-4 and Comparative Examples 2-6 significantly shortened the sleep latency of mice; compared with Example 2, Comparative Examples 1-6 significantly increased the sleep latency, indicating that the mice fell asleep more slowly, and the sample of Example 2 had the best sleep-aiding effect.

[0098] Table 4 Effects of sodium barbital on sleep latency in mice

[0099]

[0100] Note: Compared with the blank control group, +P<0.05, ++P<0.01; compared with Example 2, #P<0.05, ##P<0.01.

[0101] Combining the above three experiments, the results of samples in Examples 2-4 were all positive, indicating that peanut sprout fermentation has the effect of improving sleep; and the enzymatic hydrolysis process, enzyme preparations, and fermentation agents provided by this invention can achieve the optimal calming and sleep-aiding effect.

[0102] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing a dual-action food product that improves sleep, characterized in that, Includes the following steps: (1) Mix peanut sprout powder with water to obtain a mixture; (2) Adjust the pH of the mixture to 6-8, add enzyme preparation for enzymatic hydrolysis, and obtain the enzymatic hydrolysis solution; (3) Add carbon source and nitrogen source, and adjust the pH of the enzymatic hydrolysis solution to 6-7, and perform enzyme inactivation extraction treatment to obtain enzyme inactivation extract; (4) When the temperature of the enzyme-inactivated extract drops to 30-40℃, add the fermenting agent to carry out fermentation and obtain the fermentation liquid; (5) The fermentation broth is centrifuged, and the supernatant is vacuum dried to obtain the fermentation product; The enzyme preparation includes cellulase and bromelain; the fermentation agent includes *Lactobacillus plantarum* FTCM001 and *Lactobacillus plantarum* XD087.

2. The preparation method according to claim 1, characterized in that, The weight ratio of the fermenting agent to peanut sprout powder is 1:(50-150); the weight ratio of *Lactobacillus plantarum* FTCM001 to *Lactobacillus plantarum* XD087 is (2-3):(1-2).

3. The preparation method according to claim 1, characterized in that, The weight ratio of the enzyme preparation to peanut sprout powder is 1:(25-45); the weight ratio of the cellulase to bromelain is (2-5):(1-2).

4. The preparation method according to claim 1, characterized in that, In step (1), the weight ratio of peanut sprout powder to water is 1:(3-8).

5. The preparation method according to claim 1, characterized in that, The enzymatic hydrolysis temperature in step (2) is 45-60℃ and the enzymatic hydrolysis time is 45-70min.

6. The preparation method according to claim 1, characterized in that, In step (3), the enzyme inactivation extraction time is 15-40 min and the enzyme inactivation extraction temperature is 80-120℃.

7. The preparation method according to claim 1, characterized in that, The fermentation time in step (4) is 12-20 hours and the fermentation temperature is 30-40℃.

8. The preparation method according to claim 1, characterized in that, In step (5), the vacuum degree of vacuum drying is -0.085MPa, the drying temperature is 40-50℃, and the drying time is 6-7h.

9. The dual-element food product obtained by the preparation method according to any one of claims 1-8.

10. The application of the dual-action food according to claim 9 in the preparation of products for relieving insomnia symptoms.

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