CN115211515A - Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage and anti-depression application thereof - Google Patents

Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage and anti-depression application thereof Download PDF

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CN115211515A
CN115211515A CN202211019677.0A CN202211019677A CN115211515A CN 115211515 A CN115211515 A CN 115211515A CN 202211019677 A CN202211019677 A CN 202211019677A CN 115211515 A CN115211515 A CN 115211515A
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lactobacillus delbrueckii
bulgaricus
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马明磊
夏越
刘雪春
王艳
韩文立
李海舰
杨小宁
隋岩
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Shandong Sunflower Bioengineering Co ltd
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Abstract

The invention relates to the technical field of microorganisms, and in particular relates to a lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented beverage and an anti-depression application thereof. The fermented beverage comprises the following raw materials in parts by weight: 70-90 parts of purified water, 1-2 parts of licorice extract, 2-4 parts of buckwheat extract, 1-3 parts of red date powder, 0.5-1 part of red sage root extract, 0.5-1 part of mint extract, 0.5-1 part of cassia seed extract, 0.5-1 part of kudzu root extract, 0.5-1 part of angelica dahurica extract, 2-3 parts of lily extract, 2-3 parts of yam powder and 1-2 parts of sea buckthorn extract; the fermented beverage is obtained by fermenting Lactobacillus delbrueckii subsp bulgaricus SF-L-18. By taking the Lactobacillus delbrueckii subspecies Bulgaria SF-L-18 fermented beverage, the beverage can sooth liver and relieve depression, calm heart and tranquilize mind, and has good antidepressant effect and safety.

Description

Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage and anti-depression application thereof
Technical Field
The invention relates to the technical field of microorganisms, and in particular relates to a lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage and an anti-depression application thereof.
Background
Depression is a typical condition of depressive disorders, with the core symptoms mainly being mood depression, loss of interest, and lack of energy. Patients with depressive disorders are often associated with other cognitive, physiological, and behavioral symptoms, such as inattention, insomnia, unresponsiveness, decreased behavioral activity, and feelings of fatigue, on the basis of a depressed mood. Depression is currently treated mainly with antidepressant drugs, assisted with psychological or physical therapy. Common antidepressant drugs are divided into western medicines and traditional Chinese medicines, wherein the western medicines include fluoxetine, paroxetine, fluvoxamine, citalopram and the like, and the traditional Chinese medicines include lily and rehmannia decoction, sini powder, xiaoyao powder, bupleurum and liver soothing powder, wu sink decoction, yueju pills and the like. CN107875144B discloses an oral pharmaceutical composition for treating depression, which comprises 21-29 parts by weight of metformin hydrochloride, 6-14 parts by weight of hamamelis tannin and 11-15 parts by weight of miltefosine, and has good antidepressant effect and safety. CN110559322A discloses a sea-buckthorn compound hard capsule for treating depression and preventing anxiety and depression and a preparation method thereof, wherein the sea-buckthorn compound hard capsule comprises 15-30 parts of sea-buckthorn seed procyanidine extract, 35-45 parts of sea-buckthorn pericarp extract and 30-45 parts of ilex latifolia thunb extract in Aquifoliaceae, and is subjected to normal-temperature flash extraction and low-temperature concentration treatment to effectively treat depression. At present, pure western medicines for treating depression have strong side effects, and traditional Chinese medicine treatment has certain effect and small side effect, but has slow effect and poor taste.
Disclosure of Invention
Aiming at the problems of high side effect, slow effect and poor taste of the product in the prior art, the invention provides the lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented beverage and the anti-depression application thereof to solve the problems. By taking the lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage, the beverage can sooth liver and relieve depression, calm heart and calm nerves, and has good anti-depression effect and safety.
In a first aspect, the invention provides a lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage which comprises the following raw materials in parts by weight: 70-90 parts of purified water, 1-2 parts of licorice extract, 2-4 parts of buckwheat extract, 1-3 parts of red date powder, 0.5-1 part of red sage root extract, 0.5-1 part of mint extract, 0.5-1 part of cassia seed extract, 0.5-1 part of kudzu root extract, 0.5-1 part of angelica dahurica extract, 2-3 parts of lily extract, 2-3 parts of yam powder and 1-2 parts of sea buckthorn extract; the fermented beverage is obtained by fermenting with Lactobacillus delbrueckii subsp. Bulgaricus SF-L-18; the Lactobacillus delbrueckii subspecies Bulgaria SF-L-18 has been preserved in the China general microbiological culture Collection center in 21.10.2020, with the preservation number as follows: CGMCC No.20928, preservation address: xilu No. 1 Hospital No. 3, beijing, chaoyang, north; and is disclosed in chinese patent application CN114456977 a.
Preferably, the feed comprises the following raw materials in parts by weight: 81.5 parts of purified water, 1 part of licorice extract, 3 parts of buckwheat extract, 2 parts of red date powder, 0.5 part of salvia miltiorrhiza extract, 0.5 part of mint extract, 0.5 part of cassia seed extract, 0.5 part of kudzu root extract, 0.5 part of angelica dahurica extract, 2 parts of lily extract, 2 parts of yam powder and 1 part of sea buckthorn extract.
Preferably, the feed also comprises the following raw materials in parts by weight: 1-3 parts of honey and 2-4 parts of carrot concentrated juice.
Preferably, the amount of the Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 in the fermented beverage is 10-30 hundred million/mL.
In a second aspect, the invention provides a preparation method of the lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented beverage, which comprises the following steps:
(1) Mixing purified water, a licorice extract, a buckwheat extract, red date powder, a red sage extract, a mint extract, a cassia seed extract, a kudzuvine root extract, an angelica dahurica extract, a lily extract, yam powder and a sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, sterilizing at 120 ℃ under high pressure, and cooling to obtain a sterilized solution;
(3) Inoculating Lactobacillus delbrueckii subspecies Bulgaria SF-L-18 into MRS liquid culture medium, and culturing at 37 deg.C to obtain inoculant;
(4) Inoculating the inoculant into a sterilized solution, and culturing at 37 ℃ to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine for wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing the wall-broken fermentation liquor with honey and carrot juice, seasoning, and performing pasteurization to obtain the Lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product.
Preferably, the method comprises the following steps:
(1) Firstly, mixing purified water, a liquorice extract, a buckwheat extract, red date powder, a salvia miltiorrhiza extract, a mint extract, a cassia seed extract, a kudzuvine root extract, an angelica dahurica extract, a lily extract, yam powder and a sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, carrying out high-pressure sterilization at 120 ℃ for 30min, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus SF-L-18 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, and performing wall breaking treatment under the pressure of 50MPa to obtain wall-broken fermentation liquor;
(6) Mixing the wall-broken fermentation liquor with honey and carrot juice, seasoning, and performing pasteurization to obtain the Lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product.
Preferably, the MRS liquid medium comprises the following components: 10g of peptone, 5g of beef powder, 4g of yeast powder and K 2 HPO 4 ·7H 2 O2 g, triammonium citrate 2g, glucose 20g, CH 3 COONa·3H 2 O 5g、 MgSO 4 ·7H 2 O 0.2g、MnSO 4 ·4H 2 O0.05 g, tween 80 1mL, and distilled water 1000mL.
In a third aspect, the invention relates to application of the lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented beverage in preparing an anti-depression product.
The invention has the beneficial effects that:
the lactobacillus delbrueckii subspecies bulgaricus SF-L-18 for fermentation has strong gastrointestinal adaptability, can effectively regulate the health of gastrointestinal flora, and has the effect of improving the immunity of organisms.
The lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermentation product provided by the invention has the effects of soothing the liver, relieving depression, calming the heart and soothing the nerves, and has good prevention and treatment effects on patients with mild and moderate depression.
The lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermentation product has the following effects:
licorice root: has the effects of invigorating spleen and replenishing qi, clearing away heat and toxic materials, eliminating phlegm and stopping cough, relieving spasm and pain, and harmonizing the medicines.
Buckwheat: promoting appetite, relaxing bowel, descending qi and removing food retention;
red dates: sweet in taste, warm in nature, and capable of entering spleen and stomach channels, and has the functions of tonifying middle-jiao and Qi, nourishing blood and tranquilization and moderating drug property;
red sage root: has the effects of promoting blood circulation, removing blood stasis, dredging channels, relieving pain, clearing away heart-fire, relieving restlessness, cooling blood and eliminating carbuncle;
mint: disperse wind-heat, clear head and eyes, relieve sore throat, promote eruption, soothe liver and promote qi circulation;
cassia seed: bitter, sweet and salty taste, slightly cold nature, liver heat clearing, eyesight improving, diuresis inducing, constipation relieving, blood pressure lowering, and blood lipid reducing;
kudzu root: sweet, pungent and cool; has the effects of relieving muscles, allaying fever, promoting eruption, promoting the production of body fluid and quenching thirst;
radix angelicae: relieving exterior syndrome, dispelling cold, dispelling pathogenic wind, relieving pain, dredging nasal orifice, eliminating dampness, relieving leukorrhagia, relieving swelling, expelling pus, dispelling pathogenic wind, and relieving itching.
Lily: sweet taste and cold nature; nourishing yin and moistening lung, clearing heart fire and tranquilizing;
chinese yam: invigorating spleen, tonifying lung, reinforcing kidney, replenishing vital essence;
sea-buckthorn: invigorating spleen to promote digestion, relieving cough and eliminating phlegm, promoting blood circulation and removing blood stasis;
honey: has the effects of regulating and nourishing spleen and stomach.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The MRS solid plate culture medium used by the invention comprises the following components:
10g of peptone, 5g of beef powder, 4g of yeast powder and K 2 HPO 4 ·7H 2 O2 g, triammonium citrate 2g, glucose 20g, CH 3 COONa·3H 2 O 5g、MgSO 4 ·7H 2 O 0.2g、MnSO 4 ·4H 2 0.05g of O, 1mL of Tween 80, 15.0g of agar and 1000mL of distilled water.
The MRS liquid culture medium used in the invention comprises the following components:
10g of peptone, 5g of beef powder, 4g of yeast powder and K 2 HPO 4 ·7H 2 O2 g, triammonium citrate 2g, glucose 20g, CH 3 COONa·3H 2 O 5g、MgSO 4 ·7H 2 O 0.2g、MnSO 4 ·4H 2 0.05g of O, 1mL of Tween 80 and 1000mL of distilled water.
The preparation method of the extracts of the substances used in the invention comprises the steps of cleaning liquorice, buckwheat, salvia miltiorrhiza, mint, cassia seed, kudzuvine root, angelica dahurica, lily and sea buckthorn, crushing the cleaned liquorice, buckwheat, salvia miltiorrhiza, mint, cassia seed, kudzuvine root, angelica dahurica, lily and sea buckthorn into coarse particles, decocting the coarse particles with 10 times of water for 2 times respectively, wherein the two times of decoction are 3 hours and 1 hour, combining the two times of filtrate, standing the mixture for 2 hours, filtering the mixture to obtain an extracting solution, concentrating the extracting solution to 1.10 times (70 ℃) under the condition of vacuum degree of-0.06 Mpa, then carrying out spray drying, and finally crushing the extracting solution into 80-mesh fine powder to obtain the extract;
the preparation method of the red date powder comprises removing core of red date, adding water, pre-boiling to obtain red date juice, crushing the red date juice, adding 0.01wt% of pectinase for enzymolysis (30 ℃,80 min), heating and concentrating the enzymolysis product in vacuum to 20% of the original weight to obtain concentrated red date pulp, and then carrying out two-stage high-pressure homogenization treatment and spray drying to obtain the red date powder;
the preparation method of the yam powder comprises the steps of selecting fresh iron stick yam which is smooth, has no scab and is straight and smooth in strip shape, cleaning and peeling, cutting into slices with the thickness of 0.2-0.3 cm, immediately soaking in 0.5% sodium bisulfite aqueous solution, fishing out after 3h, cleaning with clear water, blanching in boiling water for 8min, fishing out yam slices, rinsing with clear water to remove mucus, drying the yam slices, and grinding into powder by an electric grinder to obtain the yam powder;
the preparation method of radix Dauci Sativae concentrated juice comprises pulping peeled radix Dauci Sativae Ding Yuzhu for 5min, extracting juice from the pulp with horizontal screw machine, concentrating to 40% with two-effect vacuum evaporator, sterilizing at high temperature, and cooling to obtain radix Dauci Sativae concentrated juice;
the above Glycyrrhrizae radix extract, semen Fagopyri Esculenti extract, saviae Miltiorrhizae radix extract, herba Menthae extract, semen Cassiae extract, radix Puerariae extract, radix Angelicae Dahuricae extract, bulbus Lilii extract, fructus Hippophae extract, and radix Dauci Sativae concentrated solution can also be prepared by other methods or purchased corresponding commercially available products.
Example 1
A preparation method of a lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage comprises the following steps:
(1) Firstly, 90 parts of purified water, 2 parts of licorice extract, 2 parts of buckwheat extract, 1 part of red date powder, 0.8 part of salvia miltiorrhiza extract, 0.7 part of mint extract, 0.7 part of cassia seed extract, 0.8 part of kudzu root extract, 0.8 part of angelica dahurica extract, 2.5 parts of lily extract, 2.5 parts of yam powder and 1.5 parts of sea buckthorn extract are mixed to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, carrying out high-pressure sterilization at 120 ℃ for 30min, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus SF-L-18 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, setting the pressure at 50MPa, and performing wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing and flavoring the wall-broken fermentation liquor, 2 parts of honey and 3 parts of carrot concentrated juice, and carrying out pasteurization to obtain the lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product. The number of the Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 in the fermented beverage is 15 hundred million/mL.
Example 2
A preparation method of a lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage comprises the following steps:
(1) Firstly, mixing 70 parts of purified water, 1.5 parts of licorice extract, 4 parts of buckwheat extract, 3 parts of red date powder, 1 part of salvia miltiorrhiza extract, 1 part of mint extract, 1 part of cassia seed extract, 1 part of kudzu root extract, 1 part of angelica dahurica extract, 3 parts of lily extract, 3 parts of yam powder and 2 parts of sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, sterilizing for 30min at 120 ℃ under high pressure, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus SF-L-18 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, setting the pressure at 50MPa, and performing wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing and flavoring the wall-broken fermentation liquor, 3 parts of honey and 4 parts of carrot concentrated juice, and carrying out pasteurization to obtain the lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product. The amount of the Lactobacillus delbrueckii subsp bulgaricus SF-L-18 in the fermented beverage is 18 hundred million/mL.
Example 3
A preparation method of a lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage comprises the following steps:
(1) Firstly, mixing 81.5 parts of purified water, 1 part of licorice extract, 3 parts of buckwheat extract, 2 parts of red date powder, 0.5 part of salvia miltiorrhiza extract, 0.5 part of mint extract, 0.5 part of cassia seed extract, 0.5 part of kudzu root extract, 0.5 part of angelica dahurica extract, 2 parts of lily extract, 2 parts of yam powder and 1 part of sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, sterilizing for 30min at 120 ℃ under high pressure, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus SF-L-18 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20 hours to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, setting the pressure at 50MPa, and performing wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing and flavoring the wall-broken fermentation liquor with 1 part of honey and 3 parts of carrot concentrated juice, and carrying out pasteurization to obtain the lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product. The number of the Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 in the fermented beverage is 22 hundred million/mL.
Comparative example 1
A preparation method of a fermented beverage comprises the following steps:
(1) Firstly, 85.5 parts of purified water, 2 parts of red date powder, 0.5 part of red sage root extract, 0.5 part of mint extract, 0.5 part of cassia seed extract, 0.5 part of kudzu root extract, 0.5 part of angelica dahurica extract, 2 parts of lily extract, 2 parts of yam powder and 1 part of sea buckthorn extract are mixed to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, carrying out high-pressure sterilization at 120 ℃ for 30min, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus SF-L-18 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20 hours to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, setting the pressure at 50MPa, and performing wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing and flavoring the wall-broken fermentation liquor, 1 part of honey and 3 parts of carrot concentrated juice, and carrying out pasteurization to obtain a fermentation product.
Comparative example 2
A preparation method of a fermented beverage comprises the following steps:
(1) Firstly, 83 parts of purified water, 1 part of licorice extract, 3 parts of buckwheat extract, 2 parts of red date powder, 0.5 part of salvia miltiorrhiza extract, 0.5 part of mint extract, 2 parts of lily extract, 2 parts of yam powder and 1 part of sea buckthorn extract are mixed to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, sterilizing for 30min at 120 ℃ under high pressure, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus SF-L-18 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, setting the pressure at 50MPa, and performing wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing and flavoring the wall-broken fermentation liquor, 1 part of honey and 3 parts of carrot concentrated juice, and carrying out pasteurization to obtain a fermentation product.
Comparative example 3
A preparation method of Lactobacillus delbrueckii subspecies bulgaricus CICC 20271 fermented beverage comprises the following steps:
(1) Firstly, mixing 81.5 parts of purified water, 1 part of licorice extract, 3 parts of buckwheat extract, 2 parts of red date powder, 0.5 part of salvia miltiorrhiza extract, 0.5 part of mint extract, 0.5 part of cassia seed extract, 0.5 part of kudzu root extract, 0.5 part of angelica dahurica extract, 2 parts of lily extract, 2 parts of yam powder and 1 part of sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, sterilizing for 30min at 120 ℃ under high pressure, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus CICC 20271 into MRS liquid culture medium with the inoculation amount of 1%, and culturing at 37 ℃ for 20h to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, setting the pressure at 50MPa, and performing wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing and flavoring the wall-broken fermentation liquor with 1 part of honey and 3 parts of carrot concentrated juice, and carrying out pasteurization to obtain the lactobacillus delbrueckii subspecies bulgaricus CICC 20271 fermentation product.
Comparative example 4
A preparation method of a Lactobacillus delbrueckii subspecies bulgaricus CICC 20353 fermented beverage comprises the following steps:
(1) Firstly, mixing 81.5 parts of purified water, 1 part of licorice extract, 3 parts of buckwheat extract, 2 parts of red date powder, 0.5 part of salvia miltiorrhiza extract, 0.5 part of mint extract, 0.5 part of cassia seed extract, 0.5 part of kudzu root extract, 0.5 part of angelica dahurica extract, 2 parts of lily extract, 2 parts of yam powder and 1 part of sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, carrying out high-pressure sterilization at 120 ℃ for 30min, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus CICC 20353 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20h to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, and performing wall breaking treatment under the pressure of 50MPa to obtain wall-broken fermentation liquor;
(6) Mixing and flavoring the wall-broken fermentation liquor with 1 part of honey and 3 parts of carrot concentrated juice, and carrying out pasteurization to obtain the lactobacillus delbrueckii subspecies bulgaricus CICC 20353 fermented product.
Test example 1
(1) Configuration of simulated gastric fluid: dissolving pepsin in sterilized normal saline (0.9% w/v, pH 3.0) to make pepsin final concentration be 3g/L, filtering with sterile 0.22 μm filter membrane, and using it as it is;
(2) Preparation of simulated intestinal fluid: dissolving trypsin in sterilized normal saline (0.9% w/v, pH 8.0) to make trypsin final concentration be 1g/L, adding bile salt to make bile salt final concentration be 0.3%, filtering with 0.22 μm sterile filter membrane, and using;
(3) Respectively continuously activating Lactobacillus delbrueckii subspecies Bulgaria SF-L-18, lactobacillus delbrueckii subspecies Bulgaria CICC 20271 and Lactobacillus delbrueckii subspecies Bulgaria CICC 20353 for three generations (each generation is 18 h), measuring OD value of the strain with the A600 of the strain with the three generations continuously activated, and calculating to obtain OD 5 The required amount of bacterial liquid;
(4) Will OD 5 Centrifuging the bacterial solution at 8000G for 10min, discarding supernatant, resuspending in 1mL simulated gastric juice, culturing at 37 deg.C for 3h, and counting viable bacteria on plate;
(5) Taking 8000G bacteria liquid treated by simulated gastric juice, centrifuging for 10min, discarding supernatant, suspending in simulated intestinal fluid with the same volume, culturing at 37 ℃ for 4h, and counting viable bacteria on a flat plate.
The survival rate of each strain after tolerating gastrointestinal fluids was calculated according to the following formula, and the gastrointestinal adaptability of each strain was analyzed.
Survival after tolerating gastrointestinal fluids (%) = (number of live bacteria in bacterial suspension/original number of live bacteria in bacterial suspension after tolerating simulated intestinal fluids) × 100%.
The detection results are detailed in table 1:
TABLE 1 Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 gastrointestinal tract suitability test
Strain Survival rate (%)
Lactobacillus delbrueckii subspecies bulgaricus CICC 20271 8.8
Lactobacillus delbrueckii subspecies bulgaricus CICC 20353 9.2
Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 10.3
As shown in Table 1, lactobacillus delbrueckii subspecies bulgaricus SF-L-18 of the present invention has stronger gastrointestinal adaptability than other Lactobacillus.
Test example 2
Suitability after one week of feeding, 72 SPF-grade Balb-c healthy male mice were randomly divided into 6 groups of 12 mice each, and divided into control group, example 3 group, comparative example 1 group, comparative example 2 group, comparative example 3 group, and comparative example 4 group. By constructing a chronic unpredictable emergency model, mice were given different stimuli each day, 2 were randomly selected each day: crowding (4 h), odor stimulation (12 h), ice-water bath (3 min), fasting (12 h), water deprivation (12 h), cage pouring (12 h), day and night reversing (12 h), tail clamping (2 min), squirrel cage shaking (10 min), solitary (3 h), and wet padding (12 h) for 1 week.
Control group: the basic daily feed is continuously fed for 4 weeks, and 10mL of normal saline is infused per patient.
Example 3 group: the basic daily feed is continuously fed for 4 weeks, and 10 mL/L of the Lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product prepared in example 3 is administered daily.
Comparative example 1 group: the fermented product of Lactobacillus delbrueckii subsp bulgaricus prepared in comparative example 1 was administered daily for 4 weeks with a continuous basal diet of 10 mL/dose.
Comparative example 2 group: the basic daily feed is continuously fed for 4 weeks, and 10 mL/L of the Lactobacillus delbrueckii subsp bulgaricus fermented product prepared in comparative example 2 is administered every day.
Comparative example 3 group: the basic daily feed is continuously fed for 4 weeks, and 10 mL/L of the Lactobacillus delbrueckii subsp bulgaricus fermented product prepared in comparative example 3 is administered every day.
Comparative example 4 group: the fermented product of Lactobacillus delbrueckii subsp bulgaricus prepared in comparative example 4 was administered daily for 10 mL/dose with basal diet continuously for 4 weeks.
The test method comprises the following steps:
(1) Tail suspension experiment: groups of mice were subjected to tail overhang testing in the morning of the last 1d of week 5. Fixing 1/3 of the tip of the tail of the mouse in a tail suspension box to enable the animal to be in a suspended upside-down state, and blocking the sight of the mouse by using partition plates at two sides of the suspension. Animals struggle to overcome the abnormal posture, but stop struggling when feeling unexpected. The experiment was carried out for a total of 6min and the immobility time within 4min after the recording was recorded.
(2) Forced swimming experiment: each group of mice was subjected to forced swim test in the afternoon of the last 1d of week 5. The mice were placed in a plastic cup with a water depth of 10cm for forced swimming, observed for 6min, and the time for remaining immobile 4min after recording. The test results are given in table 2 below:
TABLE 2 influence of Lactobacillus delbrueckii subspecies bulgaricus fermentation product of the present invention on depression model mice
Figure RE-GDA0003846673060000121
Note: p <0.05 compared to control group.
As can be seen from the test results shown in Table 2, compared with the control group, the Lactobacillus delbrueckii subsp. bulgaricus SF-L-18 fermented product provided by the invention can reduce the cumulative immobile time of the tail suspension and the immobile time of forced swimming of the mouse, wherein the groups of example 3 and comparative example 4 have statistical significance (p is less than 0.05), which indicates that the Lactobacillus delbrueckii subsp. bulgaricus fermented product has a certain antidepressant effect.
Test example 3
By adopting a completely random method, 60 patients with mild and moderate depression are selected and randomly divided into 5 groups, namely, a group of example 3, a group of comparative example 1, a group of comparative example 2, a group of comparative example 3 and a group of comparative example 4, wherein 12 patients in each group are added with 10ml of lactobacillus delbrueckii subsp bulgaricus fermentation products on the basis of normal treatment and 2 times a day. Treatment was continued for 8 weeks. The efficacy on disease was assessed on the HAMD scale.
HAMD reduction rate = (pre-treatment score-post-treatment score)/pre-treatment score × 100%.
The test results are shown in table 3 below:
TABLE 3 clinical test results for Lactobacillus delbrueckii subspecies Bulgaricus SF-L-18 fermentation products
Figure RE-GDA0003846673060000122
As can be seen from the test results shown in Table 3, the treatment effect was improved to some extent by administering the fermented product of Lactobacillus delbrueckii subsp. Bulgaricus SF-L-18 based on the normal treatment, and particularly the treatment effect of example 3 was more significant.
Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention.

Claims (8)

1. The lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage is characterized by comprising the following raw materials in parts by weight: 70-90 parts of purified water, 1-2 parts of licorice extract, 2-4 parts of buckwheat extract, 1-3 parts of red date powder, 0.5-1 part of red sage root extract, 0.5-1 part of mint extract, 0.5-1 part of cassia seed extract, 0.5-1 part of kudzu root extract, 0.5-1 part of angelica dahurica extract, 2-3 parts of lily extract, 2-3 parts of yam powder and 1-2 parts of sea buckthorn extract; the fermented beverage is obtained by fermenting Lactobacillus delbrueckii subsp. Bulgaricus SF-L-18;
the Lactobacillus delbrueckii subspecies Bulgaria SF-L-18 has been preserved in China general microbiological culture Collection center in 21.10.2020, with the preservation numbers: CGMCC No.20928, preservation address: xilu No. 1 Hospital No. 3, beijing, chaoyang, north.
2. The lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented beverage according to claim 1, comprising the following raw materials in parts by weight: 81.5 parts of purified water, 1 part of licorice extract, 3 parts of buckwheat extract, 2 parts of red date powder, 0.5 part of salvia miltiorrhiza extract, 0.5 part of mint extract, 0.5 part of cassia seed extract, 0.5 part of kudzu root extract, 0.5 part of angelica dahurica extract, 2 parts of lily extract, 2 parts of yam powder and 1 part of sea buckthorn extract.
3. The lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented beverage of claim 1, further comprising the following raw materials in parts by weight: 1-3 parts of honey and 2-4 parts of carrot concentrated juice.
4. The lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage of claim 1, wherein the amount of lactobacillus delbrueckii subspecies bulgaricus SF-L-18 in the fermented beverage is 10 to 30 hundred million/mL.
5. A method for preparing a lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented beverage according to claim 1, comprising the steps of:
(1) Mixing purified water, a licorice extract, a buckwheat extract, red date powder, a red sage extract, a mint extract, a cassia seed extract, a kudzuvine root extract, an angelica dahurica extract, a lily extract, yam powder and a sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, sterilizing at 120 ℃ under high pressure, and cooling to obtain a sterilized solution;
(3) Inoculating Lactobacillus delbrueckii subspecies Bulgaria SF-L-18 into an MRS liquid culture medium, and culturing at 37 ℃ to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution, and culturing at 37 ℃ to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine for wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing the wall-broken fermentation liquor with honey and carrot juice, seasoning, and performing pasteurization to obtain a lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product.
6. The method of claim 5, comprising the steps of:
(1) Firstly, mixing purified water, a licorice extract, a buckwheat extract, red date powder, a red sage root extract, a mint extract, a cassia seed extract, a kudzuvine root extract, an angelica dahurica extract, a lily extract, yam powder and a sea buckthorn extract to obtain mixed slurry;
(2) Conveying the mixed slurry into a sterilization container, carrying out high-pressure sterilization at 120 ℃ for 30min, and then cooling to 30 ℃ to obtain a sterilization solution;
(3) Inoculating lactobacillus delbrueckii subspecies bulgaricus SF-L-18 into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing at 37 ℃ for 20 hours to obtain an inoculant;
(4) Inoculating the inoculant into a sterilized solution with the inoculation amount of 1%, and culturing at 37 ℃ for 48h to obtain a fermentation liquid;
(5) Conveying the fermentation liquor to a high-pressure spray wall breaking machine, setting the pressure at 50MPa, and performing wall breaking treatment to obtain wall-broken fermentation liquor;
(6) Mixing the wall-broken fermentation liquor with honey and carrot juice, seasoning, and performing pasteurization to obtain a lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermentation product.
7. The preparation method according to claim 5, wherein the MRS liquid medium comprises the following components: 10g of peptone, 5g of beef powder, 4g of yeast powder and K 2 HPO 4 ·7H 2 O2 g, triammonium citrate 2g, glucose 20g, CH 3 COONa·3H 2 O 5g、MgSO 4 ·7H 2 O 0.2g、MnSO 4 ·4H 2 O0.05 g, tween 80 1mL, and distilled water 1000mL.
8. Use of the lactobacillus delbrueckii subsp bulgaricus SF-L-18 fermented drink according to claim 1 for preparing an antidepressant product.
CN202211019677.0A 2022-08-24 2022-08-24 Lactobacillus delbrueckii subspecies bulgaricus SF-L-18 fermented beverage and anti-depression application thereof Pending CN115211515A (en)

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