CN116035069A - Lactobacillus paracasei K56 for regulating sleep and application thereof - Google Patents

Lactobacillus paracasei K56 for regulating sleep and application thereof Download PDF

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CN116035069A
CN116035069A CN202310062183.9A CN202310062183A CN116035069A CN 116035069 A CN116035069 A CN 116035069A CN 202310062183 A CN202310062183 A CN 202310062183A CN 116035069 A CN116035069 A CN 116035069A
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lactobacillus paracasei
milk
composition
use according
sleep
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洪维鍊
赵雯
刘伟贤
侯保朝
刘彪
陈青山
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Inner Mongolia Yili Industrial Group Co Ltd
Inner Mongolia Dairy Technology Research Institute Co Ltd
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Inner Mongolia Dairy Technology Research Institute Co Ltd
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Abstract

The invention provides lactobacillus paracasei K56 for regulating sleep and application thereof. The invention firstly provides application of lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56 in preparing a composition with sleep regulating effect, wherein the lactobacillus paracasei has a preservation number of CGMCC No.15139 or DSM27447. The invention discovers that the lactobacillus paracasei strain fermentation emulsion has the application of regulating sleeping and other directions, and can be widely applied to the fields of food, medicine, feed, chemical industry and the like.

Description

Lactobacillus paracasei K56 for regulating sleep and application thereof
The invention relates to Lactobacillus paracasei K56 for resisting oxidation and regulating blood pressure and application thereof, which are divisional applications with the application number of 202011348882.2 and the application date of 2020, 11 and 26.
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a related application of lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56 (with a preservation number of CGMCC No.15139 or DSM 27447) in preparing a composition with antioxidant, blood pressure regulating and/or sleep regulating effects by fermentation.
Background
Aging is the biggest killer of human organs, and free radicals (free radicals) generated by the body are important factors for organ aging. When the body's regulated free radical is lost or unbalanced, excessive free radical production will destroy cellular DNA, alter intracellular protein structure, attack cell membrane and ultimately result in somatic cell death. Skin aging is also caused by a rapid increase in the amount of active oxygen induced by long-term exposure to Ultraviolet (UV) light or heat source (heat exposure), and causes skin dullness, skin cancer, and other diseases.
Hypertension, hyperlipidemia and hyperglycemia are collectively called as 'three highs', and are cardiovascular and cerebrovascular diseases which are harmful to human health. Experts speculate that the number of people with global hypertension in 2025 will be over 15 billion. Effective treatment or prevention of hypertensive disorders has become a worldwide health problem. At present, related researches and products of fermented dairy products with blood pressure reducing effect are mainly focused on simple addition and application of functional component substances.
Since 90 s of the last century, antihypertensive researches on microorganisms, particularly lactic acid bacteria, have been highly emphasized, and related literature reports that lactic acid bacteria can reduce serum cholesterol concentration, increase SOD activity in serum, reduce peroxidized lipid level, and have auxiliary functions of reducing blood pressure and blood lipid and resisting oxidation, but the auxiliary functions of reducing blood pressure and blood lipid and resisting oxidation of lactic acid bacteria have differences of strains and even strains, and the efficacy of lactic acid bacteria in specific fermented dairy products is to be further examined.
Disclosure of Invention
It is an object of the present invention to provide a novel use of lactobacillus paracasei K56.
Lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) strain K56 was stored in german collection for microorganisms and cell cultures (German Collection of Microorganisms and Cell Cultures) at month 6 and 27 of 2013 under accession number DSM27447; in addition, lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) K56 strain was also preserved in China general microbiological culture Collection center (China General Microbiological Culture Collection Center, CGMCC) at 12 and 29 days of 2017, with a preservation number of CGMCC No.15139.
The invention discovers that lactobacillus paracasei (Lactobacillus paracasei subsp.paracasei) K56 takes feed liquid (milk matrix) mainly containing milk components as a fermentation substrate, and substances such as superoxide dismutase, angiotensin converting enzyme inhibitors, gamma-aminobutyric acid, conjugated linoleic acid, B vitamins and the like are contained in the fermentation substrate, so that the lactobacillus paracasei (Lactobacillus paracasei subsp.paracasei) K56 has excellent total antioxidant (T-AOC) capability and has the effects of resisting oxidation, regulating blood pressure and/or regulating sleep.
Thus, the present invention provides the use of lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) having the preservation number CGMCC No.15139 or DSM27447 for the preparation of a composition having antioxidant, blood pressure regulating and/or sleep regulating effects.
According to a specific embodiment of the present invention, lactobacillus paracasei has antioxidant, blood pressure regulating and/or sleep regulating effects by fermentation to produce superoxide dismutase, angiotensin converting enzyme inhibitors, gamma aminobutyric acid, conjugated linoleic acid and/or B vitamins.
According to a specific embodiment of the present invention, the lactobacillus paracasei of the present invention is used for preparing a composition having antioxidant, blood pressure regulating and/or sleep regulating effects, which may be a food composition, a feed composition, a cosmetic composition or a pharmaceutical composition.
According to a specific embodiment of the present invention, the use of lactobacillus paracasei CGMCC No.15139 or DSM27447 for fermentation, the fermentation product is prepared, and the obtained fermentation product is used as or further for the preparation of the composition having antioxidant, blood pressure regulating and/or sleep regulating effects.
According to a specific embodiment of the present invention, the fermentation substrate used in the fermentation may be an emulsion. Preferably, the emulsion is fresh milk or reconstituted milk, and can be whole milk, low fat milk or skim milk. The emulsion may optionally contain 5% to 10% sucrose.
According to a specific embodiment of the invention, the fermentation conditions are: fermenting for 1-7 days at 35-45 ℃.
According to a specific embodiment of the invention, lactobacillus paracasei is inoculated in the form of a seed solution into a fermentation substrate, and the inoculation amount is 2-5%. The preparation of the seed liquid may be carried out with reference to conventional techniques in the probiotic field.
According to a specific embodiment of the present invention, the milk substrate used as a fermentation substrate for lactobacillus paracasei K56 of the present invention may be an emulsion having a content of 80% or more of cow's milk (whole milk, low fat milk or skim milk), and optionally, if necessary, auxiliary materials such as a sweetener and a stabilizer may be added thereto, and may be, for example, fermentation substrates commonly used in the production of fermented milk or milk drinks, and fermentation of the milk substrate by lactobacillus paracasei K56 of the present invention can achieve production of superoxide dismutase, angiotensin converting enzyme inhibitors, gamma-aminobutyric acid, conjugated linoleic acid, B vitamins and the like.
Superoxide dismutase (Superoxide Dismutase, SOD) is an antioxidant metalloenzyme existing in organisms, can catalyze superoxide anion free radical disproportionation to generate oxygen and hydrogen peroxide, and plays a vital role in organism oxidation and antioxidant balance.
Angiotensin Converting Enzyme (ACE), also known as kallikrein ii or peptidyl-carboxypeptidase, belongs to vascular endothelial cell membrane-binding enzymes. The ACE has the main functions of: catalyzing the conversion of angiotensin I to angiotensin II; inactivating the bradykinin. Angiotensin converting enzyme is an ideal target for treating diseases such as hypertension due to the two functions.
Gamma-aminobutyric acid (GABA) is a four-carbon, non-protein amino acid, which is an important inhibitory neurotransmitter, and has various physiological functions of tranquillizing, lowering blood pressure, improving sleep, etc. on mammals. It has been reported that GABA acts as a regulator of oxidative metabolites and acts as an antioxidant, and an Arabidopsis SSADH mutant grows under high temperature, and the accumulation of active oxygen intermediates (reactive oxygen intermediate, ROI) is found to lead plants to die, and GABA can reduce the accumulation of the ROI to lead organisms to avoid oxidative damage and peroxidation and decay caused by high temperature.
Conjugated Linoleic Acid (CLA) is a series of double bond linoleic acid, and has the effects of scavenging free radicals, enhancing oxidation resistance and immunity of human body, regulating blood cholesterol and triglyceride level, preventing atherosclerosis, promoting lipolysis, and performing comprehensive benign regulation on human body. It has been reported that CLA can effectively exert the function of blood vessel scavenger, remove the garbage in blood vessels, effectively regulate blood viscosity, dilate and relax vascular smooth muscle, inhibit blood movement center, reduce peripheral resistance of blood circulation, lower blood pressure, especially lower diastolic blood pressure, and improve microcirculation and stabilize blood pressure.
The antioxidant efficacy of B vitamins is well known, and B vitamins also help to clear liver fat, lower cholesterol, and prevent arteriosclerosis. In addition, vitamin B can prevent sunburn and skin cancer, keep skin smooth and moist, delay appearance of wrinkles, and promote healthy hair growth. Has research report that vitamin B in B vitamins 2 Vitamin B 3 Vitamin B 6 The components are involved in the oxidation-reduction process of the organism to different extents, and play an important role in the in-vivo oxidation-resistant defense system.
According to a specific embodiment of the present invention, superoxide dismutase, angiotensin converting enzyme inhibitors, gamma aminobutyric acid, conjugated linoleic acid and/or vitamins of the B-group in the supernatant of the K56 fermented emulsion is measured as or further used for preparing said composition with antioxidant, blood pressure regulating and/or sleep regulating effects.
According to some embodiments of the invention, the lactobacillus paracasei K56 fermented emulsion of the invention has a total antioxidant capacity (T-AOC) of 1.0 to 3.0U/mL and a slow decrease in total antioxidant capacity as the fermentation process proceeds.
According to some specific embodiments of the invention, the lactobacillus paracasei K56 fermented emulsion has total superoxide dismutase (T-SOD) activity reaching 20-30U/mL.
According to some embodiments of the invention, the lactobacillus paracasei K56 fermented emulsion according to the invention has an angiotensin converting enzyme inhibition (ACE) of 10 to 70%.
According to some embodiments of the invention, the gamma-aminobutyric acid content (GABA) in the lactobacillus paracasei K56 fermented emulsion of the invention can reach 14-21 mug/mL.
According to some specific embodiments of the invention, conjugated Linoleic Acid (CLA) in the lactobacillus paracasei K56 fermented emulsion of the invention can reach 0.01-0.06 mg/kg.
According to some embodiments of the invention, vitamin B in the Lactobacillus paracasei K56 fermented emulsion of the invention 1 The content can reach 30-70 mug/mL; vitamin B 2 The content can reach 2.8-3 mug/mL; vitamin B 3 50-80 mug/mL; vitamin B 6 The content is 8-20 mug/mL; vitamin B 9 The content of (C) is 1.6-1.8 mug/mL.
According to a specific embodiment of the present invention, the fermented product obtained by fermenting the milk substrate with lactobacillus paracasei K56 of the present invention may be directly used as a food (e.g., fermented milk, milk drink, milk powder, solid beverage, etc.), daily chemicals (cosmetics, etc.), feeds (including pet foods or probiotic products for livestock), or medicines, etc., or further used for preparing such foods, daily chemicals, feeds or pharmaceutical products. The method for preparing food, daily chemicals, feed or pharmaceutical products from the fermentation product may be performed with reference to the prior art in the field.
In a specific embodiment of the invention, the K56 fermentation product of the invention is used as or further for the preparation of a food product, preferably a fermented milk or a fermented milk beverage. Other materials commonly used in fermented milk or fermented milk beverages, such as appropriate amounts of sweeteners, stabilizers, nutritional additives, etc., may also be included in the emulsion as a fermentation substrate. The fermented milk or fermented milk beverage can be fermented milk or fermented milk beverage of active bacteria stored at low temperature, or sterilized fermented milk or fermented milk beverage stored at normal temperature. In some more specific embodiments of the present invention, the lactobacillus paracasei CGMCC No.15139 is present in the fermented milk or fermented milk beverage in an amount of 1 x 10 7 cfu/mL～1×10 11 cfu/mL。
In summary, the invention provides a new application of lactobacillus paracasei K56, and discovers that the single-strain fermented milk substrate of the strain can produce substances such as superoxide dismutase, angiotensin converting enzyme inhibitors, gamma-aminobutyric acid, conjugated linoleic acid, B vitamins and the like, and has the potential of resisting oxidation, regulating blood pressure and regulating sleep. The lactobacillus paracasei K56 strain can be used for production and application of fermented milk, milk beverage, milk powder, solid beverage, daily chemicals and the like with the effects of resisting oxidation, regulating blood pressure and regulating sleep.
Detailed Description
In order to more clearly understand the technical features, objects and advantages of the present invention, the following detailed description will be made with reference to specific embodiments thereof, and it should be understood that these examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. In the examples, each of the starting reagent materials is commercially available, and the experimental methods without specifying the specific conditions are conventional methods and conventional conditions well known in the art, or according to the conditions recommended by the instrument manufacturer.
Example 1: measurement of growth dynamics characteristics of lactobacillus paracasei K56 fermented milk
1. Experimental strains
Lactobacillus paracasei K56 (CGMCC No. 15139) used in the technical proposal of the invention;
control strain 1: lactobacillus paracasei strain l.caii 431;
control strain 2: lactobacillus paracasei strain LPC-37.
2. Activation of strains and seed liquid culture
(1) Strain activation: activating Lactobacillus paracasei with MRS culture medium, collecting strain growing to logarithmic phase, and preserving glycerol tube for use. Wherein MRS medium (g/L): 10.0g of peptone; 5.0g of yeast extract; glucose 20.0g; 5.0g of anhydrous sodium acetate; 2.0g of citric acid dihydro amine; tween-80.0 ml; 2.0g of dipotassium hydrogen phosphate; 0.2g of magnesium sulfate heptahydrate; manganese sulfate 0.05g; 10.0g of beef extract; 15g-20g of agar.
(2) Determination of growth dynamics of strain
Dissolving 12% full-fat milk powder and 6% white granulated sugar in water at normal temperature, sterilizing at 95deg.C for 5min, cooling to 36+ -1deg.C, inoculating strain as milk matrix, fermenting and culturing for 120 hr, sampling at different fermentation time periods, and measuring viable count and pH value.
Detecting the number of viable bacteria: after 25ml of the sample was mixed with 75ml of physiological saline uniformly, the plate dilution method was used for counting.
Acidity detection: the sample pH was measured using a pH meter and recorded. 10g of fermented dairy product is taken, added with 2 drops of phenolphthalein solution, and evenly mixed, and 0.1M standard NaOH solution is titrated to pink. Titration acidity calculation formula:
°T＝C×V×100/(m×0.1)。
the measurement results are shown in tables 1 and 2, respectively.
TABLE 1 colony counts of Lactobacillus paracasei in milk matrix at different fermentation times
Figure BDA0004061383430000061
From the analysis of the test data, the pH value of the K56 fermentation increases rapidly at 28h, and then decreases to be stable. The viable count of the control strain 1 and the control strain 2 increases rapidly at 36 hours, and then decreases to a stable state. K56 was analyzed p >0.05 with control strain 1, control strain 2, no significant differences; control strain 1 and control strain 2 were analyzed for p >0.05 without significant differences. Wherein the blank group is milk matrix without bacteria liquid.
TABLE 2 pH determination of Lactobacillus paracasei in milk base at different fermentation times
Figure BDA0004061383430000062
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Figure BDA0004061383430000071
From the analysis of the test data, K56, control strain 1 and control strain 2 gradually decrease within 12-36 hours. K56 was very significantly different from control strain 2 analysis p < 0.01; k56 was analyzed p >0.05 with control strain 1 without significant differences; control strain 1 and control strain 2 were analyzed for p >0.05 without significant differences. Due to temperature, the blank group contains non-killed microorganisms, which cause milk spoilage and drop in pH. Wherein the blank group is milk matrix without bacteria liquid.
(3) Preparation of strain seed solution: dissolving 12% full-fat milk powder and 6% white granulated sugar in water at normal temperature, sterilizing at 95deg.C for 5min, cooling to 36+ -1deg.C, inoculating strain, fermenting at 36+ -1deg.C for 48 hr+ -2 hr, and taking the strain as seed solution.
3. Detection of antioxidant capacity of lactobacillus paracasei fermented milk
And (3) preparing fermented milk: adding sucrose accounting for 6.5% of the total weight of the raw milk, stirring, heating to 95 ℃, sterilizing for 5min, cooling the sterilized emulsion to 37 ℃, adding seed liquid of each bacterium (lactobacillus paracasei K56 inoculation amount is 3%, and the inoculation amount of control bacterium is equivalent to K56 according to the number of viable bacteria), fermenting each sample at 37 ℃ for 24, 48, 72, 96 and 120h, centrifuging the fermented milk at 8000r/min for 15min at 4 ℃, collecting fermentation supernatant, filtering with a 0.22 mu m filter membrane, and measuring the antioxidant capacity of the filtrate.
(1) Determination of Total antioxidant (T-AOC) Capacity
In the present invention, the antioxidant capacity of Lactobacillus paracasei is mainly evaluated by measuring the total antioxidant (T-AOC) capacity.
The total antioxidant capacity (T-AOC) is measured by using a total antioxidant capacity (T-AOC) detection kit purchased from Nanjing, and the operation is performed according to the instruction of the kit. The antioxidant substance can reduce ferric ions into ferrous ions, the ferrous ions can form firm complex with phenanthrene substances, and the oxidation resistance of the ferric ions can be measured by a colorimetric method. In the measurement process, when a sample to be measured is mixed with a reagent provided by the kit, a vortex mixer is fully and uniformly mixed, the mixture is placed in a water bath at 37 ℃ for 30 minutes for 10 minutes, the wavelength is 520nm, the optical path is 1cm, double distilled water is zeroed, and the absorbance value of each tube is measured.
Figure BDA0004061383430000081
In the present invention, the results of the T-AOC capacity measurement of the supernatants of probiotics fermentation at different fermentation times are shown in Table 3.
TABLE 3 Total antioxidant capacity
Figure BDA0004061383430000082
(2) Detection of other antioxidant-associated capacities in K56 fermentation samples
In the invention, other antioxidant related capacities in K56 fermentation samples, including total superoxide dismutase (T-SOD) activity, angiotensin Converting Enzyme (ACE) inhibition rate, superoxide anion free radical scavenging capacity, OH free radical scavenging capacity, DPPH free radical scavenging capacity, gamma-aminobutyric acid content (GABA), conjugated Linoleic Acid (CLA) and B vitamins content of supernatant fluid with different fermentation time are also detected.
In the invention, the activity of total superoxide dismutase (T-SOD) is measured by using a total superoxide dismutase (T-SOD) kit purchased from Nanjing, and the operation is carried out according to the instruction of the kit. The superoxide anion free radical is generated by a xanthine and xanthine oxidase reaction system, hydroxylamine is oxidized to form nitrite, the nitrite is mauve under the action of a color developing agent, and the absorbance of the nitrite is measured by a visible spectrophotometer. When the detected sample contains SOD, the specific inhibition effect on superoxide anion free radical is realized, so that the formed nitrite is reduced, the absorbance value of the measuring tube is lower than that of the control tube during colorimetric, and the activity of the SOD in the sample can be obtained through calculation.
Figure BDA0004061383430000083
The method for measuring Angiotensin Converting Enzyme (ACE) mainly comprises a colorimetry method, an enzyme coupling method and the like. In the present invention, the Angiotensin Converting Enzyme (ACE) inhibitory activity was measured by ultraviolet spectrophotometry: taking a 2mL centrifuge tube, adding 50 mu L of 50mmol/L sodium borate buffer solution with 5mmol/L Hippuriyl-His-Leu (HHT) as a substrate, adding 20 mu L of liquid to be detected, mixing, preheating for 3min in a 37 ℃ water bath, adding 10 mu L of ACE enzyme solution, reacting for 30min at 37 ℃, and adding 100 mu L of 1mol/L hydrochloric acid to terminate the reaction. 1.7mL of ethyl acetate was added, 15s was shaken, centrifuged at 3000 Xg for 10min, and the upper 1mL ethyl acetate layer (layer where the sample was located) was separated. The organic solvent was removed by evaporation under heating, 1mL of deionized water was added thereto, and the hippuric acid was completely dissolved by shaking, and the absorbance of the sample was measured at 228nm by a micro-ultraviolet spectrophotometry. The ACE inhibition ratio (ACEI) is calculated as follows:
Figure BDA0004061383430000091
wherein A represents the absorbance value of the reaction of ACE and HIL to generate hippuric acid completely (control); b represents the absorbance value (sample) of hippuric acid generated after the liquid to be tested reacts with ACE and HIL; c represents the absorbance of hippuric acid generated after the reaction of ACE and HIL added with hydrochloric acid.
In the invention, the determination of the scavenging capacity of superoxide anion free radicals adopts a pyrogallol-spectrophotometry method. The pyrogallol-spectrophotometry is widely applied to the primary screening of antioxidants in the food and pharmaceutical industries and the evaluation of the antioxidant function of various active substances for scavenging superoxide anion free radicals. The pyrogallol autoxidation process is chain reaction, can produce superoxide anion free radical, its content of its own oxidation products can be detected with the spectrophotometry, the antioxidation ability of the antioxidant substance of indirect evaluation through this method. The method comprises the following specific steps: 0.88mL of 0.1mol/L Tris-HCl buffer (pH 8.2) was taken in a test tube, and 0.2mL of 1.0mmol/L ethylenediamine tetraacetic acid (EDTA), 0.2mL of sample, and 0.3mL of distilled water were sequentially added. The mixture was reacted in a water bath at 25℃for 10 minutes, then 0.4mL of 9.0mmol/L of pyrogallol was added thereto, and after 60 minutes of exact reaction, 100. Mu.L of 12.0mol/L of HCl was added thereto to terminate the reaction. The above were all averaged 3 times in parallel and the experiment was repeated 3 times. The calculation formula is as follows: clearance = (Ac-As)/ac×100%, where As: measuring absorbance at a wavelength of 325 nm; ac: the blank tube was replaced with 0.2mL of distilled water, and absorbance was measured using the same sample tube as the operation method.
In the invention, the OH free radical scavenging ability is determined by the phenanthroline method.
In the present invention, the method for measuring the gamma-aminobutyric acid (GABA) content is carried out according to "QB/T4587-2013 gamma-aminobutyric acid".
In the invention, the method for measuring the content of the Conjugated Linoleic Acid (CLA) is carried out according to a gas chromatography NY/T1671-2008 for measuring the content of the Conjugated Linoleic Acid (CLA) in milk and dairy products.
In the invention, the content of the B vitamins is measured by adopting a high performance liquid chromatograph, and specific operating conditions are carried out by referring to ' HPLC for detecting water-soluble vitamins in Collybia albuminosa ' (Li Yan, food research and development, 2018 ').
In the present invention, the measurement results of total superoxide dismutase (T-SOD) activity, angiotensin Converting Enzyme (ACE) inhibition ratio, superoxide anion free radical scavenging ability, OH free radical scavenging ability, DPPH free radical scavenging ability, gamma-aminobutyric acid content (GABA), conjugated Linoleic Acid (CLA) and B vitamins of the fermentation supernatants of probiotics with different fermentation times of K56 are shown in Table 4.
TABLE 4 determination of antioxidant Capacity associated with K56 fermentation supernatant
Figure BDA0004061383430000101
From the measurement results, the K56 fermented emulsion has excellent antioxidant capacity and can be used for production and application of fermented milk, milk beverage, milk powder, solid beverage, daily chemicals and the like with antioxidant, blood pressure regulating and sleep regulating effects.
EXAMPLE 2 Lactobacillus paracasei K56 fermentation preparation of milk beverage with antioxidant, blood pressure regulating and sleep regulating potential
The embodiment provides the milk beverage prepared by fermenting the lactobacillus paracasei K56 and having the potential of resisting oxidization, regulating blood pressure and regulating sleep. The preparation method of the beverage comprises the following steps:
(a) Inoculating lactobacillus paracasei K56 into skimmed milk for fermentation, and obtaining fermented milk after fermentation;
(b) Blending sweetener and/or sour agent to form blending liquid, mixing the blending liquid with the fermented milk prepared in step (a), homogenizing, and preparing into fermented milk beverage.
Further, the fermentation temperature in the step (a) is 37 ℃ and the fermentation time is 72 hours.
Further, in the step (b), the mixing volume ratio of the fermented milk to the blending liquid is 1:4.
Further, the homogenizing pressure in the step (b) is 30MPa.
Further, in the step (b), a sterilization process may be used, and the preferred sterilization temperature is 120 ℃ and the sterilization time is 15min.
The beverage of the embodiment has good potential of resisting oxidation, regulating blood pressure and regulating sleep in two products at low temperature and normal temperature.

Claims (10)

1. Use of lactobacillus paracasei (Lactobacillus paracasei subsp. Paracasei) having the accession number CGMCC No.15139 or DSM27447 for the preparation of a composition having sleep-modulating effect.
2. Use according to claim 1, wherein lactobacillus paracasei has sleep-modulating effects by fermentation to produce superoxide dismutase, angiotensin converting enzyme inhibitors, gamma aminobutyric acid, conjugated linoleic acid and/or B vitamins;
preferably, lactobacillus paracasei further has antioxidant and/or blood pressure regulating effects as an active substance.
3. The use according to claim 1, wherein the composition is a food composition, a feed composition, a cosmetic composition or a pharmaceutical composition.
4. Use according to any one of claims 1-3, wherein the emulsion is fermented with lactobacillus paracasei CGMCC No.15139 or DSM27447 to prepare a fermented product, which is used as or further for preparing the composition with sleep-modulating effect.
5. The use according to claim 4, wherein the emulsion is fresh cow's milk or reconstituted milk, optionally containing 5% to 10% sucrose.
6. The use according to claim 5, wherein the fermentation conditions are: fermenting for 1-7 days at 35-45 ℃.
7. Use according to claim 5 or 6, wherein lactobacillus paracasei is inoculated in the emulsion in the form of a seed solution in an amount of 2% to 5%.
8. The use according to claim 5, wherein the supernatant of the fermentation product is used as or further for the preparation of the composition with sleep-modulating effect.
9. The use according to claim 4, wherein the composition having sleep-regulating effect is fermented milk, fermented milk beverage, milk powder or solid beverage.
10. Use according to claim 9, wherein the lactobacillus paracasei CGMCC No.15139 is present in the fermented milk or fermented milk beverage in an amount of 1 x 10 7 cfu/mL～1×10 11 cfu/mL。
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