CN116195631A - Lactobacillus paracasei ET-22 for regulating sleep and application thereof - Google Patents

Lactobacillus paracasei ET-22 for regulating sleep and application thereof Download PDF

Info

Publication number
CN116195631A
CN116195631A CN202310062047.XA CN202310062047A CN116195631A CN 116195631 A CN116195631 A CN 116195631A CN 202310062047 A CN202310062047 A CN 202310062047A CN 116195631 A CN116195631 A CN 116195631A
Authority
CN
China
Prior art keywords
lactobacillus paracasei
milk
composition
use according
sleep
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202310062047.XA
Other languages
Chinese (zh)
Inventor
赵雯
洪维鍊
刘伟贤
侯保朝
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Inner Mongolia Yili Industrial Group Co Ltd
Inner Mongolia Dairy Technology Research Institute Co Ltd
Original Assignee
Inner Mongolia Yili Industrial Group Co Ltd
Inner Mongolia Dairy Technology Research Institute Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Inner Mongolia Yili Industrial Group Co Ltd, Inner Mongolia Dairy Technology Research Institute Co Ltd filed Critical Inner Mongolia Yili Industrial Group Co Ltd
Priority to CN202310062047.XA priority Critical patent/CN116195631A/en
Publication of CN116195631A publication Critical patent/CN116195631A/en
Pending legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00Milk preparations; Milk powder or milk powder preparations
    • A23C9/12Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • A23C9/1234Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00Milk preparations; Milk powder or milk powder preparations
    • A23C9/16Agglomerating or granulating milk powder; Making instant milk powder; Products obtained thereby
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38Other non-alcoholic beverages
    • A23L2/382Other non-alcoholic beverages fermented
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/385Concentrates of non-alcoholic beverages
    • A23L2/39Dry compositions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66Microorganisms or materials therefrom
    • A61K35/74Bacteria
    • A61K35/741Probiotics
    • A61K35/744Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747Lactobacilli, e.g. L. acidophilus or L. brevis
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99Cosmetics 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
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00Drugs for disorders of the nervous system
    • A61P25/20Hypnotics; Sedatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00General protective or antinoxious agents
    • A61P39/06Free radical scavengers or antioxidants
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00Drugs for disorders of the cardiovascular system
    • A61P9/12Antihypertensives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Food Science & Technology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Nutrition Science (AREA)
  • Epidemiology (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Anesthesiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Toxicology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Birds (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Cardiology (AREA)
  • Gerontology & Geriatric Medicine (AREA)
  • Dermatology (AREA)
  • Physiology (AREA)
  • Animal Husbandry (AREA)
  • Zoology (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention provides lactobacillus paracasei ET-22 for regulating sleep and application thereof. The invention firstly provides application of lactobacillus paracasei (Lactobacillus paracasei) ET-22 in preparing a composition with sleep regulating effect, wherein the preservation number of the lactobacillus paracasei is CGMCC No.15077. 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 ET-22 for regulating sleep and application thereof
The invention relates to lactobacillus paracasei ET-22 with the application number of 202011348850.2, the application date of 2020, 11 months and 26 days and the name of antioxidant and blood pressure regulating and application of the lactobacillus paracasei ET-22.
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a related application of lactobacillus paracasei (Lactobacillus paracasei) ET-22 (preservation number CGMCC No. 15077) 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 lactobacillus paracasei ET-22.
Lactobacillus paracasei (Lactobacillus paracasei) ET-22 strain was stored in China General Microbiological Culture Center (CGMCC) (address: institute of microbiology, national academy of sciences of china) at 12 months 18 of 2017: lactobacillus paracasei (Lactobacillus paracasei); the preservation number is CGMCC No.15077.
The invention discovers that lactobacillus paracasei (Lactobacillus paracasei) ET-22 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, conjugated linoleic 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 invention provides the use of lactobacillus paracasei (Lactobacillus paracasei) with the preservation number of CGMCC No.15077 for preparing a composition with the effects of resisting oxidation, regulating blood pressure and/or regulating sleep.
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, in the application of the present invention, lactobacillus paracasei CGMCC No.15077 is utilized for fermentation to prepare a fermentation product, and the obtained fermentation product is used as or further used for preparing the composition with the effects of resisting oxidation, regulating blood pressure and/or regulating sleep.
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 ET-22 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 beverage, and fermentation of the milk substrate by lactobacillus paracasei ET-22 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, B vitaminsThe biotin also helps to remove liver fat, reduce 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 ET-22 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 ET-22 fermented emulsion of the invention has a total antioxidant capacity (T-AOC) of 1.0 to 2.7U/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 ET-22 fermentation emulsion has total superoxide dismutase (T-SOD) activity reaching 20-30U/mL.
According to some embodiments of the invention, the lactobacillus paracasei ET-22 fermented emulsion of the invention has an angiotensin converting enzyme inhibition (ACE) of 10 to 50%.
According to some embodiments of the invention, the gamma-aminobutyric acid content (GABA) in the lactobacillus paracasei ET-22 fermented emulsion of the invention can reach 10-17 mug/mL.
According to some specific embodiments of the invention, conjugated Linoleic Acid (CLA) in the lactobacillus paracasei ET-22 fermented emulsion of the invention can reach 0.01-0.07 mg/kg.
According to some embodiments of the invention, vitamin B in the lactobacillus paracasei ET-22 fermented emulsion of the invention 1 The content can reach 60-80 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 7-14 mug/mL; vitamin B 9 The content of (C) is 2.5-2.9 mug/mL.
According to a specific embodiment of the present invention, the fermented product obtained by fermenting the milk substrate with lactobacillus paracasei ET-22 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.), feed (including pet food or probiotic products for livestock), or pharmaceuticals, etc., or further used for preparing such food, daily chemicals, feed 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 present invention, the ET-22 fermentation product of the present 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.15077 is present in the fermented milk or fermented milk beverage in an amount of 1×10 6 cfu/mL~1×10 11 cfu/mL。
In summary, the invention provides a new application of lactobacillus paracasei ET-22, and discovers that the strain single-strain fermented milk matrix can generate 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 ET-22 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 ET-22 fermented milk
1. Experimental strains
Lactobacillus paracasei ET-22 (CGMCC No. 15077) used in the technical scheme 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 BDA0004061364950000061
From the analysis of the test data, the ET-22 grows rapidly in the number of live bacteria at 28h, and then reduces 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. ET-22 was significantly different from control strain 1, control strain 2 (p < 0.05); 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 BDA0004061364950000062
/>
Figure BDA0004061364950000071
From the analysis of the test data, the pH value of the ET-22 fermentation gradually decreases within 28-72 h, and the pH values of the control strain 1 and the control strain 2 gradually decrease within 12-36 h. ET-22 was very significantly different from control strain 1, control strain 2 analysis p < 0.01; 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 ℃, 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 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 BDA0004061364950000081
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 BDA0004061364950000082
(2) Detection of other antioxidant-associated capacities in ET-22 fermentation samples
In the invention, other antioxidant related capacities in ET-22 fermentation samples are also detected, 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.
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 BDA0004061364950000083
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 BDA0004061364950000091
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 invention, the measurement results of total superoxide dismutase (T-SOD) activity, angiotensin Converting Enzyme (ACE) inhibition rate, 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 ET-22 are shown in table 4.
TABLE 4ET-22 determination of antioxidant Capacity in relation to fermentation supernatant
Figure BDA0004061364950000101
The measurement results show that the ET-22 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 ET-22 fermentation to prepare milk beverage with antioxidant, blood pressure regulating and sleep regulating potential
The embodiment provides the lactobacillus paracasei ET-22 fermented milk beverage with the potential of resisting oxidation, regulating blood pressure and regulating sleep. The preparation method of the beverage comprises the following steps:
(a) Inoculating lactobacillus paracasei ET-22 into skim 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 96-120 h.
Further, in the step (b), the mixing volume ratio of the fermented milk to the blending liquid is 1:2-4.
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) having the accession number CGMCC No.15077 for the preparation of a composition having sleep-modulating effects.
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 claim 1, wherein the emulsion is fermented with lactobacillus paracasei CGMCC No.15077 to prepare a fermented product, the fermented 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, 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.15077 is present in the fermented milk or fermented milk beverage in an amount of 1 x 10 6 cfu/mL~1×10 11 cfu/mL。
CN202310062047.XA 2020-11-26 2020-11-26 Lactobacillus paracasei ET-22 for regulating sleep and application thereof Pending CN116195631A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202310062047.XA CN116195631A (en) 2020-11-26 2020-11-26 Lactobacillus paracasei ET-22 for regulating sleep and application thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202011348850.2A CN113349254B (en) 2020-11-26 2020-11-26 Antioxidant and blood pressure regulating lactobacillus paracasei ET-22 and application thereof
CN202310062047.XA CN116195631A (en) 2020-11-26 2020-11-26 Lactobacillus paracasei ET-22 for regulating sleep and application thereof

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN202011348850.2A Division CN113349254B (en) 2020-11-26 2020-11-26 Antioxidant and blood pressure regulating lactobacillus paracasei ET-22 and application thereof

Publications (1)

Publication Number Publication Date
CN116195631A true CN116195631A (en) 2023-06-02

Family

ID=77524548

Family Applications (3)

Application Number Title Priority Date Filing Date
CN202011348850.2A Active CN113349254B (en) 2020-11-26 2020-11-26 Antioxidant and blood pressure regulating lactobacillus paracasei ET-22 and application thereof
CN202310017824.9A Pending CN116195630A (en) 2020-11-26 2020-11-26 Lactobacillus paracasei ET-22 for regulating blood pressure and application thereof
CN202310062047.XA Pending CN116195631A (en) 2020-11-26 2020-11-26 Lactobacillus paracasei ET-22 for regulating sleep and application thereof

Family Applications Before (2)

Application Number Title Priority Date Filing Date
CN202011348850.2A Active CN113349254B (en) 2020-11-26 2020-11-26 Antioxidant and blood pressure regulating lactobacillus paracasei ET-22 and application thereof
CN202310017824.9A Pending CN116195630A (en) 2020-11-26 2020-11-26 Lactobacillus paracasei ET-22 for regulating blood pressure and application thereof

Country Status (1)

Country Link
CN (3) CN113349254B (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113892651A (en) * 2021-10-08 2022-01-07 河北一然生物科技股份有限公司 Novel application of composite probiotic preparation in antioxidant field
CN114752525B (en) * 2022-04-21 2022-09-09 广西壮族自治区水牛研究所 Lactobacillus casei with blood pressure lowering effect and application thereof
CN114903083B (en) * 2022-06-09 2023-10-20 西安百跃羊乳集团有限公司 Bioactive peptide probiotic fermented sheep milk and preparation method thereof
CN115851520B (en) * 2022-11-09 2024-04-19 厦门元之道生物科技有限公司 Lactobacillus paracasei and application thereof

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005102559A (en) * 2003-09-29 2005-04-21 Domer Inc NEW STRAIN AND METHOD FOR MASS-PRODUCING gamma-AMINOBUTYRIC ACID(GABA) USING THE SAME
KR20100117918A (en) * 2009-04-27 2010-11-04 강원대학교산학협력단 The preparing method for natural gaba using serial fermentation of plant originated lactic acid bacteria with gaba-producing ability and protease-producing ability, and the food and beverage containing the natural gaba
KR20110052808A (en) * 2009-11-13 2011-05-19 충북대학교 산학협력단 A polysaccharide producing lactobacillus paracasei and a use thereof
CN104855956A (en) * 2015-04-13 2015-08-26 劲膳美生物科技股份有限公司 Medical formula food for insomnia
KR20160006377A (en) * 2014-07-09 2016-01-19 주식회사 유담 yogurt
CN110964653A (en) * 2018-09-30 2020-04-07 内蒙古伊利实业集团股份有限公司 Lactobacillus paracasei ET-22 capable of adjusting intestinal flora balance

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140328814A1 (en) * 2008-05-13 2014-11-06 Oü Tervisliku Piima Biotehnoloogiate Arenduskeskus (Bio-Competence Centre of Antimicrobial and antihypertensive probiotic composition, food product and dietary supplement comprising microorganism strain lactobacillus plantarum tensia dsm 21380 and method for suppressing contaminating microbes in a food product
CN103060243A (en) * 2013-01-17 2013-04-24 福建省农业科学院农业工程技术研究所 Sub-lactobacillus casei and sub-cheese subspecies strain
ITUB20152376A1 (en) * 2015-07-07 2017-01-09 Alfasigma Spa LACTOBACILLUS PARACASEI FOR THE PRODUCTION OF CONJUGATED LINOLEIC ACID, NUTRITIONAL AND PHARMACEUTICAL PREPARATIONS THAT INCLUDE IT AND THEIR USE
CN105400727A (en) * 2015-12-21 2016-03-16 南昌大学 Lactobacillus paracasei with antioxidant activity and application thereof
CN106071436A (en) * 2016-06-12 2016-11-09 中国食品发酵工业研究院 A kind of Germinatus Phragmitis fermented beverage that can improve sleep quality and preparation method thereof
CN108283285A (en) * 2017-12-31 2018-07-17 深圳市金乐智能健康科技有限公司 A kind of health-care physiotherapeutic food of diabetes
US11617772B2 (en) * 2018-09-11 2023-04-04 Direct Digital Llc Nutritional supplements and therapeutic compositions comprising probiotics
CN110964658B (en) * 2018-09-30 2021-07-06 内蒙古伊利实业集团股份有限公司 Lactobacillus paracasei ET-22 with immunoregulation function
CN110960559B (en) * 2018-09-30 2021-02-26 内蒙古伊利实业集团股份有限公司 Food, oral cleaning and pharmaceutical composition for inhibiting lactic acid bacteria strain of oral pathogenic bacteria
CN110893195B (en) * 2019-09-30 2023-03-14 内蒙古伊利实业集团股份有限公司 Lactobacillus paracasei ET-22 with function of relieving intestinal inflammation
CN110903995B (en) * 2019-09-30 2023-03-24 内蒙古伊利实业集团股份有限公司 Probiotic edible composition and food with digestion promoting effect
CN111304113A (en) * 2019-12-20 2020-06-19 佛山科学技术学院 Lactobacillus paracasei subspecies paracasei and application thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005102559A (en) * 2003-09-29 2005-04-21 Domer Inc NEW STRAIN AND METHOD FOR MASS-PRODUCING gamma-AMINOBUTYRIC ACID(GABA) USING THE SAME
KR20100117918A (en) * 2009-04-27 2010-11-04 강원대학교산학협력단 The preparing method for natural gaba using serial fermentation of plant originated lactic acid bacteria with gaba-producing ability and protease-producing ability, and the food and beverage containing the natural gaba
KR20110052808A (en) * 2009-11-13 2011-05-19 충북대학교 산학협력단 A polysaccharide producing lactobacillus paracasei and a use thereof
KR20160006377A (en) * 2014-07-09 2016-01-19 주식회사 유담 yogurt
CN104855956A (en) * 2015-04-13 2015-08-26 劲膳美生物科技股份有限公司 Medical formula food for insomnia
CN110964653A (en) * 2018-09-30 2020-04-07 内蒙古伊利实业集团股份有限公司 Lactobacillus paracasei ET-22 capable of adjusting intestinal flora balance

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
PEGAH HAMZEHLOU等: "Production of Vitamins B3, B6 and B9 by Lactobacillus Isolated from Traditional Yogurt Samples from 3 Cities in Iran, Winter 2016", APPLIED FOOD BIOTECHNOLOGY, vol. 5, no. 2, 31 December 2018 (2018-12-31), pages 107 - 120 *
刘春晓等: "内蒙古锡林郭勒盟地区传统奶油制品中产共轭亚油酸乳酸菌的分离筛选与鉴定", 食品科学, vol. 37, no. 9, 15 May 2016 (2016-05-15), pages 186 - 191 *
张俊等: "副干酪乳杆菌的益生特性及其应用研究进展", 包装与食品机械, vol. 37, no. 4, 30 August 2019 (2019-08-30), pages 47 - 52 *

Also Published As

Publication number Publication date
CN113349254A (en) 2021-09-07
CN116195630A (en) 2023-06-02
CN113349254B (en) 2022-11-29

Similar Documents

Publication Publication Date Title
CN113367191B (en) Lactobacillus paracasei K56 for resisting oxidation and regulating blood pressure and application thereof
CN113349254B (en) Antioxidant and blood pressure regulating lactobacillus paracasei ET-22 and application thereof
CN113773999B (en) Lactobacillus paracasei fermentation filtrate, preparation method and application thereof
CN113350383B (en) Bifidobacterium lactis BL-99 capable of resisting oxidation and regulating blood pressure and application thereof
CN113930361B (en) Fermentation medium of lactobacillus paracasei and application thereof
Zhao et al. Preventive effect of Lactobacillus plantarum KSFY02 isolated from naturally fermented yogurt from Xinjiang, China, on d-galactose–induced oxidative aging in mice
CN109694886B (en) Green tea fermentation filtrate and preparation method and application thereof
CN111471604B (en) Application of saccharomyces cerevisiae ZLG-6 and lactobacillus plantarum Picp-2
CN108925982A (en) A kind of preparation method of high anti-oxidation activity pomegranate ferment
KR101984400B1 (en) Process for producing antioxidant and anti-fatigue efficacy of mulberry vinegar
KR20110135477A (en) Fermented liquid by saccharomyces sp. having the effects of antioxidation and whitening and method of manufacturing thereof
CN112167345B (en) Food composition and pharmaceutical composition containing antioxidant lactobacillus fermentation product
CN113632922A (en) Lysate, preparation method and application
CN114081862B (en) Preparation and application of Saccharomyces cerevisiae fermentation product from highland barley wine distiller&#39;s yeast in Pan-Himalaya region
TWI824330B (en) Jabuticaba extract fermented by lactobacillus and preparation method thereof
CN111973495A (en) Preparation method and application of fermented vegetable-berry fruit product
CN113974099B (en) Preparation method and application of blood orange fermented product with antioxidant activity
CN111500642B (en) Blueberry fermentation product and preparation method and application thereof
TWI752334B (en) Food composition and pharmaceutical composition with anti-oxidation fermentation metabolites of lactic acid bacterium
CN117223808B (en) Bigeminal live bacteria fermented beverage for high yield of gamma-aminobutyric acid
CN107048130A (en) A kind of dendrobium candidum ferment pearl powder beverage and preparation method thereof
TWI742406B (en) Anti-oxidation food composition and pharmaceutical composition with strain of lactic acid bacterium
KR102042363B1 (en) Method for producing functional fermented materials containing astaxanthin and GABA by fermentation methods
KR20220028782A (en) The fermented cornus officinalis composition for an antioxidant and antiobesity effects and method for manufacturing the same
KR20230076251A (en) Method for rice bran fermented extracted material product

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination