CN116211898A - Lactobacillus bifidus BL-99 for regulating sleep and application thereof - Google Patents
Lactobacillus bifidus BL-99 for regulating sleep and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/1203—Addition of, or treatment with, enzymes or microorganisms other than lactobacteriaceae
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/16—Agglomerating or granulating milk powder; Making instant milk powder; Products obtained thereby
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/20—Hypnotics; Sedatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
Abstract
The invention provides bifidobacterium lactis BL-99 for regulating sleep and application thereof. The invention firstly provides application of bifidobacterium lactis (Bifidobacterium lactis) BL-99 in preparing a composition with sleep regulating effect, wherein the preservation number of the bifidobacterium lactis is CGMCC No.15650. The invention discovers that the bifidobacterium lactis 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
The invention relates to bifidobacterium lactis BL-99 with the application number of 202011348864.4, the application date of 2020, 11 months and 26 days and the name of 'antioxidation and blood pressure regulation' and application thereof.
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a related application of bifidobacterium lactis (Bifidobacterium lactis) BL-99 (preservation number CGMCC No. 15650) in fermentation preparation of a composition with antioxidant, blood pressure regulating and/or sleep regulating effects.
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 bifidobacterium lactis BL-99.
Bifidobacterium lactis (Bifidobacterium lactis) BL-99 strain has been preserved in China general microbiological culture collection center CGMCC (address: north Chen West Lu No.1, 3 of the area of Chachiensis in Beijing, and national academy of sciences of China) at 26 th 2018, and named after classification: bifidobacterium lactis (Bifidobacterium lactis); the preservation number is CGMCC No.15650.
The invention discovers that bifidobacterium lactis (Bifidobacterium lactis) BL-99 takes feed liquid (milk matrix) mainly containing milk components as a fermentation substrate, and the fermentation product contains substances such as superoxide dismutase, angiotensin converting enzyme inhibitors, gamma-aminobutyric acid, B vitamins and the like, 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 bifidobacterium lactis (Bifidobacterium lactis) having a collection number of cgmccno.15650 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, the bifidobacterium lactis has antioxidant, blood pressure regulating and/or sleep regulating effects by fermentation to produce superoxide dismutase, angiotensin converting enzyme inhibitors, gamma aminobutyric acid and/or B vitamins.
According to a specific embodiment of the present invention, the bifidobacterium lactis of the present invention is used in the preparation of 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 bifidobacterium lactis CGMCC No.15650 for fermentation to produce a fermentation product, the fermentation product obtained being used as or further for the production 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, the bifidobacterium lactis is inoculated in the fermentation substrate in the form of a seed solution, 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 as a fermentation substrate for bifidobacterium lactis BL-99 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, for example, fermentation substrates commonly used in the production of fermented milk or milk drinks, and fermentation of these milk substrates by bifidobacterium lactis BL-99 of the present invention can achieve the production of angiotensin converting enzyme inhibitors, superoxide dismutase, gamma-aminobutyric acid, B vitamins and the like.
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.
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.
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.
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 and/or vitamins of group B in the supernatant of BL-99 fermented emulsion is measured as or further used for preparing the composition with antioxidant, blood pressure regulating and/or sleep regulating effects.
According to some embodiments of the invention, the bifidobacterium lactis BL-99 fermented emulsion of the invention has an angiotensin converting enzyme inhibition (ACE) of 10-55%.
According to some embodiments of the invention, the bifidobacterium lactis BL-99 fermented emulsion of the invention has a total antioxidant capacity (T-AOC) of 1.0-2.5U/mL and gradually decreases as the fermentation process proceeds.
According to some embodiments of the invention, the bifidobacterium lactis BL-99 fermented emulsion has a superoxide anion radical scavenging capacity of 20-40%.
According to some specific embodiments of the invention, the total superoxide dismutase (T-SOD) activity of the bifidobacterium lactis BL-99 fermentation emulsion can reach 20-30U/mL.
According to some embodiments of the invention, the gamma-aminobutyric acid content (GABA) in the bifidobacterium lactis BL-99 fermentation emulsion of the invention can reach 10-20 mug/mL.
According to some embodiments of the invention, vitamin B in the bifidobacterium lactis BL-99 fermented emulsion of the invention 1 The content can reach 60-100 mug/mL; vitamin B 2 The content can reach 2.8-3.1 mug/mL; vitamin B 3 40-60 mug/mL; vitamin B 6 The content is 9-14 mug/mL; vitamin B 9 The content of (C) is 1.5-1.6 mug/mL.
According to a specific embodiment of the present invention, the fermented product obtained by fermenting the milk substrate with bifidobacterium lactis BL-99 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 pharmaceuticals, 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 the present inventionIn a specific embodiment, the BL-99 fermentation product of the present invention is used as or further in 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 content of bifidobacterium lactis CGMCC No.15650 in the fermented milk or the fermented milk beverage is 1×10 7 cfu/mL～1×10 11 cfu/mL。
In summary, the invention provides a new application of bifidobacterium lactis BL-99, and discovers that the strain is used for fermenting milk matrix by virtue of single bacteria, can generate substances such as superoxide dismutase, angiotensin converting enzyme inhibitors, gamma-aminobutyric acid, B vitamins and the like, and has the potential of resisting oxidation, regulating blood pressure and regulating sleep. The bifidobacterium lactis BL-99 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: determination of growth dynamics characteristics of bifidobacterium lactis BL-99 fermented milk
1. Experimental strains
Bifidobacterium lactis BL-99 (CGMCC No. 15650) used in the technical scheme of the invention;
control strain 1: commercial bifidobacterium lactis strain BB-12.
2. Activation of strains and seed liquid culture
(1) Strain activation: activating bifidobacterium lactis with a BSM culture medium, taking a strain growing to a logarithmic phase, and preserving a glycerol tube for later use. Wherein BSM medium (g/L): 5.0g of peptone, 5.0g of beef extract, 10.0g of tryptone, 5.0g of yeast extract powder, 0.5% fructo-oligosaccharide, 10.0g of glucose, 1.0mL of tween-80 and K 2 HPO 4 2.0g of sodium acetate 5.0g, diammonium hydrogen citrate 2.0g, znSO 4 ·7H 2 O 0.25g，MgSO 4 ·7H 2 O 0.1g，1.0％CaCO 3 Distilled water was added (endogenous adjustment of pH) to 1000mL.
(2) 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 culturing to logarithmic phase as seed solution.
3. Detection of antioxidant capacity of fermented milk of bifidobacterium lactis
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 ℃, respectively adding various bacterial seed solutions (inoculum size is 3%), respectively fermenting each sample at 37 ℃ for 24, 48, 72, 96 and 120h, centrifuging the fermented emulsion at 8000r/min for 15min, collecting fermentation supernatant, filtering with a 0.22 μm 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 bifidobacterium lactis is mainly evaluated by measuring the inhibition rate of Angiotensin Converting Enzyme (ACE), the total antioxidant (T-AOC) capacity and the scavenging capacity of superoxide anion radicals.
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:
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.
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.
The superoxide anion radical scavenging capacity was determined using a pyrogallol-spectrophotometer 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 present invention, the results of the detection of the inhibition rate of angiotensin converting enzyme of the fermentation supernatants of probiotics at different fermentation times are shown in Table 1, the results of the measurement of the T-AOC ability are shown in Table 2, and the results of the measurement of the scavenging ability of superoxide anions are shown in Table 3.
TABLE 1 angiotensin converting enzyme inhibition ratio
TABLE 2 Total antioxidant capacity
TABLE 3 superoxide anion radical scavenging Capacity
(2) Detection of other antioxidant-associated capacities in BL-99 fermentation samples
In the invention, other antioxidant related capacities in BL-99 fermentation samples are also detected, including total superoxide dismutase (T-SOD) activity, gamma-aminobutyric acid content (GABA) and B vitamins content of supernatants at different fermentation times.
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.
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 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 invention, the determination results of total superoxide dismutase (T-SOD) activity, gamma-aminobutyric acid content (GABA) and B vitamins of the fermentation supernatants of probiotics with different fermentation times of BL-99 are shown in Table 4.
TABLE 4BL-99 fermentation supernatant-related antioxidant capacity assay
As shown by the measurement results, the BL-99 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 preparation of milk beverage with antioxidant, blood pressure and sleep-regulating potential by Bifidobacterium lactis BL-99 fermentation
The embodiment provides the milk beverage prepared by fermenting the bifidobacterium lactis BL-99 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 bifidobacterium lactis BL-99 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.
EXAMPLE 3 preparation of fermented milk with antioxidant, blood pressure and sleep potential by Bifidobacterium lactis BL-99 fermentation
Heating the defatted milk to above 50 ℃, adding 6% of white granulated sugar, stirring until the white granulated sugar is completely dissolved, preheating to 60-65 ℃, homogenizing under 20Mpa pressure, performing heat treatment at about 90 ℃ for 5-8 minutes, cooling to 37 ℃, and inoculating bifidobacterium lactis BL-99 seed liquid with the inoculum size of 2% -3%. Fermenting at 37deg.C for 6-8 hr to obtain functional fermented dairy product. The product has sour and sweet taste, good flavor, and has effects of resisting oxidation, regulating blood pressure, and regulating sleep.
Claims (10)
1. Use of bifidobacterium lactis (Bifidobacterium lactis) having a preservation number of CGMCC No.15650 for the preparation of a composition having sleep-modulating effects.
2. Use according to claim 1, wherein bifidobacterium lactis has sleep-regulating efficacy by fermentation to produce superoxide dismutase, angiotensin converting enzyme inhibitors, gamma aminobutyric acid and/or B vitamins.
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 claim 1, wherein the emulsion is fermented with bifidobacterium lactis CGMCC No.15650 to prepare a fermentation product, the fermentation product obtained being used as or further for preparing the composition with sleep-regulating effect.
5. The use according to claim 4, wherein the emulsion is fresh cow's milk or reconstituted milk, with or without sucrose in an amount of 5% to 10%.
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 bifidobacterium lactis 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 content of bifidobacterium lactis CGMCC No.15650 in the fermented milk or fermented milk beverage is 1 x 10 7 cfu/mL～1×10 11 cfu/mL。
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| CN104146254B (en) * | 2014-08-14 | 2016-06-15 | 劲膳美生物科技股份有限公司 | The full nutritional formulas of hypertension |
| CN105368738A (en) * | 2015-10-23 | 2016-03-02 | 中国农业大学 | Lactobacillus paracasei and appliance thereof |
| CN105483234A (en) * | 2015-12-26 | 2016-04-13 | 上海中优生物高科技有限责任公司 | Hypertension-type obese gene individualization intervention composition and preparation method and system |
| CN105831749A (en) * | 2016-04-15 | 2016-08-10 | 劲膳美生物科技股份有限公司 | Medical formula food for hypertension |
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| CN110964655B (en) * | 2018-09-30 | 2021-05-11 | 内蒙古伊利实业集团股份有限公司 | Bifidobacterium lactis BL-99 and application thereof |
| CN110959676B (en) * | 2018-09-30 | 2022-11-25 | 内蒙古伊利实业集团股份有限公司 | Fermented milk product containing bifidobacterium lactis and application thereof |
| CN110964657B (en) * | 2018-09-30 | 2021-03-12 | 内蒙古伊利实业集团股份有限公司 | Bifidobacterium lactis BL-99 capable of improving immunity and application thereof |
| EP3812452A4 (en) * | 2018-09-30 | 2021-11-03 | Inner Mongolia Yili Industrial Group Co., Ltd. | Bifidobacterium lactis bl-99 and application thereof |
| CN109771470A (en) * | 2019-03-21 | 2019-05-21 | 慕恩（广州）生物科技有限公司 | A kind of composition and preparation method thereof containing probiotics |
| CN110893193B (en) * | 2019-06-27 | 2023-03-24 | 内蒙古伊利实业集团股份有限公司 | Novel application of bifidobacterium lactis BL-99 |
| CN110892961B (en) * | 2019-06-27 | 2022-03-29 | 内蒙古伊利实业集团股份有限公司 | Solid beverage containing bifidobacterium lactis BL-99 and application thereof |
| CN116211898A (en) * | 2020-11-26 | 2023-06-06 | 内蒙古伊利实业集团股份有限公司 | Lactobacillus bifidus BL-99 for regulating sleep and application thereof |
| CN114470010B (en) * | 2022-02-07 | 2024-02-27 | 北京华元生物技术研究院 | Novel application of bifidobacterium lactis BL-11 |
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- 2020-11-26 CN CN202310388410.7A patent/CN116211898A/en active Pending
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