Lactobacillus rhamnosus, probiotic preparation for promoting growth and development and preparation method thereof

Abstract

The present invention provides a Lacticaseibacillus rhamnosus UP180, and its preservation number is CGMCC NO.32109. The present invention also provides a fermentation product obtained by fermenting astragalus extract by the Lacticaseibacillus rhamnosus UP180. The present invention also provides a probiotic preparation, including the Lacticaseibacillus rhamnosus UP180, which helps to promote growth and development. The present invention also provides a preparation method of the probiotic preparation, including the steps of fermenting and culturing the probiotics to obtain bacterial mud, and freeze-drying the bacterial mud after mixing it with a freeze-drying protective agent. It is convenient for subsequent industrial applications.

Description

Technical Field

[0001] The invention belongs to the field of probiotics, and specifically relates to a Lactobacillus rhamnosus and a fermentation product thereof obtained by fermentation and capable of promoting secretion of growth hormone, a probiotic preparation comprising the Lactobacillus rhamnosus and the fermentation product thereof, and a preparation method thereof. Background Art

[0002] Growth hormone is a hormone secreted by the pituitary gland and is highly related to metabolism such as bone metabolism, protein metabolism, sugar metabolism, fat metabolism, and electrolyte metabolism.

[0003] In childhood, insufficient secretion of growth hormone can lead to various problems, among which developmental disorders are the most well-known. In adulthood, insufficient secretion of growth hormone can cause metabolic abnormalities, cardiovascular disorders, etc. Even in healthy people, growth hormone secretion will gradually decrease as they age.

[0004] It is well known that compared with young people, the elderly are relatively in a state of insufficient growth hormone secretion. Some people point out that insufficient growth hormone secretion leads to a decrease in the quality of life of the elderly.

[0005] For the problem of insufficient growth hormone secretion in childhood, it has been found that developmental disorders can be improved by directly administering growth hormone. For the problem of insufficient growth hormone secretion in adulthood, the patient's metabolic abnormalities, cardiovascular disorders and other problems can also be improved by administering growth hormone. For the insufficient growth hormone secretion of the elderly, their quality of life can also be improved by administering growth hormone.

[0006] However, the use of growth hormone requires strict dosage control and professional medical supervision. In addition, growth hormone is a polypeptide that must be administered by injection rather than orally, and patient compliance is poor.

[0007] It has been reported that using an oral growth hormone secretion promoter to replace growth hormone can increase the secretion of growth hormone in the body. Therefore, finding a safe and mild growth hormone secretion promoter has become a problem that needs to be solved.

[0008] Compared with the Lactobacillus rhamnosus in the prior art, the Lactobacillus rhamnosus disclosed in the Chinese invention application with publication number CN117866831A has at least the following beneficial effects: (1) good tolerance to the acidic environment of gastric juice and bile salts; (2) good adhesion to the intestine; (3) ability to inhibit pathogenic bacteria; (4) ability to increase the expression of intestinal tight junction proteins and have the effect of preventing and / or improving intestinal epithelial barrier damage; (5) ability to significantly increase the expression of the anti-inflammatory factor IL-10 and inhibit the expression of pro-inflammatory factors, and can inhibit or improve colon inflammation and regulate immunity.

[0009] The degradation rate of Lactobacillus rhamnosus TG021 disclosed in the Chinese invention application with publication number CN117736908A in high-lipid cells is 44.44% for triglyceride TG and 26.05% for low-density lipoprotein cholesterol LDL-C.

[0010] However, the above-mentioned prior arts do not involve the problem of growth hormone promoters. Summary of the invention

[0011] The first object of the present invention is to provide a Lactobacillus rhamnosus that can ferment astragalus and promote intestinal peristalsis.

[0012] The second object of the present invention is to provide a fermentation product obtained by fermenting Astragalus with Lactobacillus rhamnosus.

[0013] The third object of the present invention is to provide a probiotic preparation comprising Lactobacillus rhamnosus.

[0014] The fourth object of the present invention is to provide a method for preparing a probiotic preparation.

[0015] The present invention is achieved through the following technical solutions:

[0016] A rhamnosus Lactobacillus UP180, whose Latin name is Lacticaseibacillus rhamnosus. The preservation number of the strain is CGMCC NO.32109. The strain was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on September 29, 2024, the preservation unit code is CGMCC, the preservation unit address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code 100101, and the preservation number is CGMCC32109. The bacteria were alive when preserved.

[0017] A fermentation product obtained by fermenting the astragalus extract with the rhamnosus Lactobacillus UP180, wherein the fermentation product comprises a growth hormone secretion promoter;

[0018] The structure of the growth hormone secretion promoter is as follows:

[0019]

[0020] A probiotic preparation comprises the Lactobacillus rhamnosus UP180 and the fermentation product.

[0021] The probiotic preparation also includes a freeze-drying protective agent.

[0022] The freeze-drying protective agent comprises 2-4% skim milk powder, 1.5-1.8% soy protein isolate, 1.5-2.5% casein, 1.5-3.5% whey protein, 3-4% L-cysteine, 1-5% oligofructose, 2-3% trehalose, 0.6-2.5% calcium alginate and the balance is water.

[0023] The preparation method of the probiotic preparation comprises the steps of fermenting and culturing the probiotics to obtain bacterial mud, mixing the bacterial mud with a freeze-drying protective agent, and then freeze-drying.

[0024] The culture medium used in the fermentation culture includes the following components in percentage by weight:

[0025] Yeast extract powder 0.4-0.8%, beef extract powder 0.4-0.7%, peptone 0.7-0.9%, diammonium hydrogen citrate 0.1-0.2%, dipotassium hydrogen phosphate 0.1-0.5%, anhydrous magnesium sulfate 0.01-0.03%, Tween 80 0.05-0.15%, glucose 1.5-2.5%, maltodextrin 1.0-1.5%, galacto-oligosaccharide 0.5-0.8%, astragalus extract 0.3-0.5%, water balance. The preparation method of the astragalus extract is as follows:

[0026] The astragalus root is crushed to obtain powder, and the powder is subjected to supercritical CO2 extraction to obtain the product.

[0027] The entrainer used in the supercritical CO2 extraction includes ethanol;

[0028] The liquid ratio of the powder to the entrainer is 3 g:1 mL.

[0029] The supercritical CO2 extraction adopts an extraction pressure of 30-50MPa;

[0030] The extraction temperature used in the supercritical CO2 extraction is 45-50°C;

[0031] The extraction time of the supercritical CO2 extraction is 1-1.5h.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] 1. The present invention provides a specific Lactobacillus rhamnosus UP180, whose preservation number is CGMCC NO.32109, which provides a new strain resource for scientific research and industrial application.

[0034] 2. The present invention provides a fermentation product obtained by fermenting astragalus extract with rhamnosus Lactobacillus UP180. The fermentation product may have unique biological activity or function, providing a possibility for developing new functional foods or drugs.

[0035] 3. The present invention provides a probiotic preparation, which contains Lactobacillus rhamnosus UP180, which helps to promote growth and development, which may have a positive effect on improving human health, promoting growth and development, etc.

[0036] 4. The present invention provides a method for preparing a probiotic preparation, comprising the steps of fermenting and culturing probiotics to obtain bacterial sludge, and mixing the bacterial sludge with a lyophilization protective agent and then freeze-drying the mixture. This method can conveniently realize the large-scale production and preservation of probiotic preparations, and provides technical support for subsequent industrial applications. DETAILED DESCRIPTION

[0037] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0038] Example 1 Astragalus Extraction

[0039] Since astragalus is used as both medicine and food, it is mainly absorbed by the human body after being fermented by probiotics in the intestines after being decocted. Therefore, screening out probiotics that can ferment astragalus from the human intestine and isolating beneficial compounds from the fermentation products has become a method for finding target bacteria.

[0040] The intestinal flora of newborn babies is relatively simple, with probiotics accounting for the majority, and they have not used antibiotics, so the activity of probiotics is very high. Therefore, the success rate of screening and isolating bacteria with target functions from the feces of newborn babies is relatively high. The specific extraction process of Astragalus extract is as follows:

[0041] Cut the Astragalus into thin slices of 0.5-0.8 cm, put them into an oven at 70°C and dry them for 4 hours, then crush them into 2-5 mm to obtain Astragalus granules for use in supercritical CO2 fluid extraction.

[0042] After the extraction kettle is installed, three layers of folded absorbent gauze are placed at one end of the extraction kettle and flattened, and then the crushed astragalus granules are placed in the extraction kettle, a certain volume of ethanol solution is poured into the extraction kettle, and then the three layers of folded absorbent gauze are placed at the other end of the extraction kettle to obtain a loaded extraction kettle. The mass ratio of the astragalus granules to the volume of the extraction kettle is 3g:1mL, and the volume concentration (v / v) of the ethanol solution is 65%.

[0043] Fix the loaded extraction kettle in the heating box, turn on the power of the supercritical CO2 fluid extraction equipment, set the temperature of the heating box, and when the temperature of the extraction kettle reaches the set value, turn on the air inlet switch to adjust the pressure. When the pressure of the extraction kettle reaches the set conditions, perform static extraction, wherein the temperature of the extraction kettle is 45-50°C, the pressure of the extraction kettle is 30-50MPa, and the time of the static extraction is 1 hour.

[0044] After the static extraction time is reached, the fine-tuning valve heating switch is turned on and the temperature is set to 90°C. After the temperature reaches the set value, the air outlet valve is opened and the gas flow in the recovery bottle is controlled by the fine-tuning valve so that the tail-connected flow meter displays a flow of 2 liters per minute. The pressure of the extraction kettle is balanced by a pneumatic pump to keep the pressure of the extraction kettle constant. The entrainer pump is turned on and the flow rate of the entrainer ethanol solution is set to 1.5 milliliters per minute. Dynamic extraction is started. After the dynamic extraction time is reached, the entrainer pump switch is turned off, the air inlet switch is turned off, the heating switch of the heating box is turned off, and CO2 in the extraction kettle is allowed to continue to be discharged. The recovery bottle continues to collect the extract until CO2 is emptied and the recovery bottle is removed, wherein the volume concentration (v / v) of the entrainer ethanol solution is 60%-70%, and the dynamic extraction time is 1-1.5 hours.

[0045] The extract in the recovery bottle is transferred to a triangular flask, and anhydrous ethanol is added to the triangular flask to obtain an alcohol precipitation solution. The alcohol precipitation solution is kept at 4° C. for 2 hours, and the alcohol precipitation solution is centrifuged using a centrifuge to collect the lower layer of white solid matter, put it into an oven, and dry it at 70° C. to obtain a dry astragalus extract, wherein the rotation speed of the centrifuge is 3000 revolutions per minute, and the centrifugation is performed at room temperature for 20 minutes.

[0046] Example 2 Screening and Identification Process of Bacteria

[0047] 1. Take 2 g of fresh feces from a healthy baby within 7 days after birth, filter it with sterile saline, spread it on a plate with MSR culture medium after gradient dilution, place it in an anaerobic incubator, and culture it in a constant temperature incubator at 37°C for 48 hours.

[0048] 2. After the colonies grow on the plate, use an inoculation loop to pick the colonies and purify them by streaking according to their color, size, transparency, edge shape, etc. Perform purification three times until the color, size, transparency, and edge shape of the colonies on the same plate are uniform.

[0049] 3. The purified colonies were subjected to Gram staining and catalase analysis, retaining Gram-positive bacteria and catalase-negative bacteria.

[0050] 4. Spread the remaining bacteria on an MRS plate containing 0.8% calcium carbonate, and pick out the slightly larger bacteria with a transparent calcium-soluble circle.

[0051] 5. The selected bacteria will be subjected to the next step of gastric acid and bile salt tolerance test.

[0052] Artificial gastric juice tolerance test: The treated bacterial liquid was inoculated into artificial gastric juice with a pH value of 3.0, and cultured in a 37°C water bath for 3 hours. The number of live bacteria was determined and the survival rate was calculated.

[0053]

[0054]

[0055] Artificial intestinal fluid tolerance test: After the bacteria with high survival rate were treated in artificial gastric fluid for 3 hours, they were inoculated into artificial intestinal fluid (containing 0.3% bile salt) and cultured in a 37°C water bath for 6 hours. After that, the number of live bacteria was determined and the survival rate was calculated.

[0056] Bacteria Survival rate (%) JYLR-985 2 UP180 94 TG021 43 IOB820 40 DY801 65 grx21 44

[0057] The above survival rate data can be used to screen out the strains UP180 and DY801 with relatively high survival rates.

[0058] 6. The strains UP180 and DY801 with high survival rates were subjected to fermentation tests. Specifically, the bacteria with high survival rates were further cultured in MRS medium (0.5wt% of astragalus extract was additionally added to the above MRS medium), with a bacterial inoculation amount of 2% and a temperature of 37°C. After testing the fermentation product of UP180 using a human growth hormone releasing hormone receptor IGF-1 (GHRH) ELISA detection kit, it was found that the fermentation product of strain UP180 had an agonistic effect on the human growth hormone releasing hormone receptor. A single colony of UP180 can be isolated from the above MRS medium.

[0059] A single colony of strain UP180 was picked and placed in Lysis Buffer for Microorganism to Direct PCR, and lysed in a water bath at 80°C for 30 min to obtain a bacterial DNA template.

[0060] Universal primers for bacterial 16S rRNA gene amplification: 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-TACGACTTAACCCCAATCGC-3'.

[0061] The reaction system (25 μL) was: 1 μL each of universal primers 27F and 1492R for bacterial 16S rRNA gene amplification, 0.5 μL of UP180 bacterial DNA template, 12.5 μL of 2×RapidTaq Master Mix, and finally dd H2O was used to make up the 25 μL reaction system.

[0062] PCR reaction program: 95°C pre-denaturation for 3 min; 95°C denaturation for 15 s, 60°C annealing for 15 s, 72°C extension for 15 s, 35 cycles; 72°C final extension for 5 min; 4°C storage. After PCR, the reaction mixture was electrophoresed in 1% agarose gel, and the electrophoresis buffer was 1×TAE (50 mmol / L Tris-acetic acid, 1 mmol / L ETA, pH 8.0).

[0063] UP180 was sequenced and compared with the sequencing results, and was identified as belonging to Lactobacillus rhamnosus. The 16s rDNA sequencing results of the bacteria are as follows:

[0064]

[0065] See SEQ ID NO.1 for details.

[0066] Example 3 Lactobacillus rhamnosus UP180 fermented Astragalus extract

[0067] (1) In order to extract, separate and identify various substances from the fermentation broth, the present invention purifies the fermentation broth. The specific steps are as follows:

[0068] Next: 1. After the fermentation is completed, the fermentation liquid is soaked in acetone, centrifuged and the bacterial residue is discarded, the acetone soaked clear liquid is collected and adsorbed on an NKA-9 macroporous adsorption resin column at a rate of 1.5 BV / h, and then eluted with 8 times the column volume of water and 60% acetonitrile-water solution, respectively, at an elution flow rate of 1.5 BV / h, to remove some impurity pigments; then eluted with 4 times the column volume of 95% acetone, collected the eluate, and concentrated to obtain a crude extract;

[0069] 2. Take the crude extract and add a small amount of methanol to dissolve it by ultrasonication, then stir and mix it with G200-300 mesh silica gel, and vacuum dry it to remove the solvent; and fill the silica gel with the crude product into the upper layer of G200-300 mesh silica gel column, and then use petroleum ether: ethyl acetate = 7:4, petroleum ether: ethyl acetate = 4:3, and petroleum ether: ethyl acetate = 3:2 as the mobile phase for gradient elution, and collect the eluent in sections;

[0070] 3. The collected eluate was tested for effective components by HPLC, and the same components were combined and concentrated under vacuum. The concentrated solution was collected and dissolved in DMSO and then loaded on the DAC dynamic axial compression system. After HPLC detection, the target products were combined and vacuum dried to obtain the pure compound.

[0071] HPLC analysis conditions: mobile phase: chromatographic grade acetonitrile (phase A), pure water containing 0.5% (v:v) formic acid (phase B); chromatographic column: ZORBAX SB-C18 (250×4.6 mm, 5 μm); flow rate: 1 mL / min; maximum pressure limit: 380 bar; detector: VWD; column temperature: 25°C; wavelength: 255 nm, 305 nm; injection volume: 20 μL; retention time: 50 min; gradient elution conditions: starting at A15% B 85%, reaching A 80% B20% at 25 min, reaching A85% B15% at 30 min, A reaching 95% B10% at 32 min, A reaching 100% at 38 min, A reaching 30% B70% at 45 min, and maintaining A20% B 80% at 50 min.

[0072] The DAC dynamic axial compression system has C18 filler, a 50% acetonitrile-water solution as the mobile phase, a controlled flow rate of 10 ml / min, and a detection wavelength of 247 nm.

[0073] When the substances extracted by the above method were studied in pharmacology, it was found that the following compounds showed the effect of promoting the secretion of growth hormone:

[0074]

[0075] The spectral data of the compound are as follows:

[0076] 1H NMR (300MHz, CD3CN, δ, ppm)

[0077] 9.58(1H in phenol),8.62(1H in sec.amide),8.23(1H in 2-pyridine),7.56(1H in 2-pyridine),6.00(1H in 2-furan),5.99(1H in 2-furan),5.61(1H in 1-ethylene),5.48(1H in 1-ethylene),5.54(1H in 1-ethylene),5.42(1H in 1-ethylene),4.38(1H in methine),3.56(1H in methine),3.46(2H in methine),3.29(2H in methylene),2.73(2H in methylene),1.63(6H in methyl),1.25(3H in methyl),1.21(3H in methyl).

[0078] 13C NMR (300MHz, CD3CN, δ, ppm)

[0079] 171.6,153.6,153.3,151.7,151.1,140.9,135.3,135.2,130.7,127.7,125.7,121.3,106.8,106.6,53.1,39.1,37.3,35.7,29.1,23.2,17.9,17.6,15.4.

[0080] Example 4 A method for preparing a probiotic freeze-dried powder

[0081] UP180 was inoculated into MRS test tube culture medium and cultured at 35-40°C for 32-38 hours. The test tube strain was then inoculated into a triangular flask liquid culture medium for fermentation and cultured at 35-40°C for 36-40 hours.

[0082] The culture medium comprises the following components in percentage by weight: 0.4-0.8% yeast extract, 0.4-0.7% beef extract, 0.7-0.9% peptone, 0.1-0.2% diammonium hydrogen citrate, 0.1-0.5% dipotassium hydrogen phosphate, 0.01-0.03% anhydrous magnesium sulfate, 0.05-0.15% Tween 80, 1.5-2.5% glucose, 1.0-1.5% maltodextrin, 0.5-0.8% galacto-oligosaccharide, 0.3-0.5% astragalus extract, and the balance of distilled water.

[0083] The fermented probiotic fermentation liquid is passed through a tubular centrifuge and centrifuged at a high speed of 8000-10000 r / min to obtain probiotic bacterial mud; the bacterial mud is mixed with a freeze-drying protective agent and freeze-dried at -60°C to obtain the probiotic bacterial mud.

[0084] The freeze-drying protective agent is 2-4% skim milk powder, 1.5-1.8% soy protein isolate, 1.5-2.5% casein, 1.5-3.5% whey protein, 3-4% L-cysteine, 1-5% oligofructose, 2-3% trehalose, 0.6-2.5% calcium alginate and the balance is water.

[0085] Example 5: Common fermentation method for preparing probiotics

[0086] UP180 was inoculated into MRS test tube culture medium and cultured at 35-40°C for 32-38 hours. The test tube strain was then inoculated into a triangular flask liquid culture medium for fermentation and cultured at 35-40°C for 36-40 hours.

[0087] The culture medium comprises the following components in percentage by weight: 0.4-0.8% yeast extract powder, 0.4-0.7% beef extract powder, 0.7-0.9% peptone, 0.1-0.2% diammonium hydrogen citrate, 0.1-0.5% dipotassium hydrogen phosphate, 0.01-0.03% anhydrous magnesium sulfate, 0.05-0.15% Tween 80, 1.5-2.5% glucose, 1.0-1.5% maltodextrin, 0.5-0.8% galacto-oligosaccharide, and the balance distilled water.

[0088] Subsequent treatment method Reference Example 6 Probiotic freeze-dried preparation method

[0089] Example 7 Characterization of the Growth Hormone Secretion Promoting Effect

[0090] Twenty-four 7-week-old male C57BL / 6 mice (Japan SLC, Inc.) were divided into three groups, each including 8 mice: a group of mice administered with a powder of a growth hormone secretion promoter (experimental group 1); a group of mice administered with a powder of a probiotic prepared by ordinary fermentation (experimental group 2); and a group of mice administered with a control nutritional composition powder (control group).

[0091] Each group was fed for 2 weeks and then blood was collected from the abdominal vein. Protease inhibitors were added to the blood at a concentration of 1 / 100 of the blood mass. The blood was centrifuged at 6000 rpm and 2°C for 2 minutes to separate the plasma and measure the concentration of growth hormone.

[0092] The acyl auxin concentration in plasma was measured by ELISA kit (auxin ELISA kit). Plasma was diluted 5-fold with assay buffer for measurement, and the acyl auxin concentration was calculated by correcting the measured value by the dilution factor.

[0093] The results were: experimental group 1: 77.6fmol / ml; experimental group 2: 58.0 and the control group: 38.4fmol / ml. It can be seen that the experimental group showed a significant increase in the concentration of active growth hormone in the blood. The specific data are as follows:

[0094] Serial number Experimental group 1 (fmol / ml) Experimental group 2 (fmol / ml) Control group (fmol / ml) 1 74.8 54.8 39.1 2 79.0 62.7 34.1 3 79.6 59.3 36.7 4 78.4 61.5 46.3 5 70.4 55.6 38.4 6 78.1 56.3 35.3 7 79.8 55.3 36.7 8 80.4 58.4 40.4

[0095] Example 8 Characterization of the ability of probiotics to promote growth and development in mice

[0096] 1. Experimental animals: Healthy SPF-grade 6-week-old Balb / c male and female mice were grouped and mixed, and the female mice were made pregnant. Newborn mice were obtained. The newborn mice were randomly divided into 3 groups, 8 in each group: control group, experimental group 1 (fed with probiotic freeze-dried powder), and experimental group 2 (fed with ordinary fermented probiotic powder).

[0097] 2. Intervention method: From the first week to the fifth week after birth, newborn mice were given normal saline (control group) and lyophilized powder of Yihe probiotics (experimental groups 1 and 2) by oral gavage. The oral dose of mice was 1×10 8 CFU / day / probiotics.

[0098] 3. Index analysis:

[0099] Determination of the average length of the left and right tibia of mice: At the end of the experiment, the length of each group of mice was measured while they were lightly anesthetized and stretched to evaluate the potential effect of probiotics on their growth and development. Results: Compared with the control group, the experimental group with probiotic intervention promoted a significant increase in the length of mice, which indicates that the probiotic agent involved in the present invention has the potential to promote bone growth and development.

[0100] Serial number Experimental group 1 (mm) Experimental group 2 (mm) Control group (mm) 1 15.23 14.12 13.01 2 15.04 13.01 13.10 3 14.91 13.89 12.99 4 14.93 13.97 12.98 5 14.85 13.65 13.06 6 14.74 13.74 12.90 7 14.68 13.57 13.03 8 14.92 13.69 13.11 average value 14.91 13.70 13.02

[0101] Mouse serum index detection: After the experiment, blood was collected from each group of mice. After the blood was left to stand for 1 hour, it was centrifuged for 10 minutes to separate the serum. The serum insulin-like growth factor 1 (IGF-1) level and serum osteocalcin (OCN) level of mice were measured by enzyme-linked immunosorbent assay (ELISA) using a kit.

[0102] Compared with the control group, the serum insulin-like growth factor 1 (IGF-1) level and serum osteocalcin (OCN) level of young mice in the probiotic intervention group (experimental group 1) were higher.

[0103]

[0104]

[0105]

[0106] This result indicates that the probiotics of the present invention may promote the expression of growth factors by activating endocrine pathways, thereby affecting bone growth and metabolism.

[0107] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

Claims

1. A Lactobacillus rhamnosus UP180, characterized in that: Its accession number is CGMCC NO.32109.

2. A fermentation product obtained by fermenting the astragalus extract with the Lactobacillus rhamnosus UP180 according to claim 1, characterized in that: The fermentation product includes a growth hormone secretion promoter; The structure of the growth hormone secretion promoter is as follows:

3. A probiotic preparation, characterized in that: The invention comprises the Lactobacillus rhamnosus UP180 as claimed in claim 1.

4. The probiotic preparation according to claim 3, characterized in that: Lyoprotectants are also included.

5. The probiotic preparation according to claim 4, characterized in that: The freeze-drying protective agent is 2-4% skim milk powder, 1.5-1.8% soy protein isolate, 1.5-2.5% casein, 1.5-3.5% whey protein, 3-4% L-cysteine, 1-5% oligofructose, 2-3% trehalose, 0.6-2.5% calcium alginate and the balance is water.

6. The method for preparing the probiotic preparation according to claim 3, characterized in that: The method comprises the steps of fermenting and culturing probiotics to obtain bacterial mud, mixing the bacterial mud with a freeze-drying protective agent, and then freeze-drying the mixture.

7. The method for preparing the probiotic preparation according to claim 6, characterized in that: The culture medium used in the fermentation culture includes the following components in percentage by weight: Yeast extract powder 0.4-0.8%, beef extract powder 0.4-0.7%, peptone 0.7-0.9%, diammonium hydrogen citrate 0.1-0.2%, dipotassium hydrogen phosphate 0.1-0.5%, anhydrous magnesium sulfate 0.01-0.03%, Tween 80 0.05-0.15%, glucose 1.5-2.5%, maltodextrin 1.0-1.5%, galacto-oligosaccharide 0.5-0.8%, astragalus extract 0.3-0.5%, and the balance is water.

8. The method for preparing the probiotic preparation according to claim 7, characterized in that: The preparation method of the astragalus extract is as follows: The astragalus root is crushed to obtain powder, and the powder is subjected to supercritical CO2 extraction to obtain the product.

9. The method for preparing the probiotic preparation according to claim 8, characterized in that: The entrainer used in the supercritical CO2 extraction includes ethanol; The liquid ratio of the powder to the entrainer is 3 g:1 mL.

10. The method for preparing the probiotic preparation according to claim 8, characterized in that: The supercritical CO2 extraction adopts an extraction pressure of 30-50MPa; The extraction temperature used in the supercritical CO2 extraction is 45-50°C; The extraction time of the supercritical CO2 extraction is 1-1.5h.

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