CN110923172B - Lactobacillus salivarius with bowel relaxing effect and cholesterol reducing effect and application thereof - Google Patents
Lactobacillus salivarius with bowel relaxing effect and cholesterol reducing effect and application thereof Download PDFInfo
- Publication number
- CN110923172B CN110923172B CN201911353237.7A CN201911353237A CN110923172B CN 110923172 B CN110923172 B CN 110923172B CN 201911353237 A CN201911353237 A CN 201911353237A CN 110923172 B CN110923172 B CN 110923172B
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- lactobacillus salivarius
- cholesterol
- strain
- hcs20
- lactobacillus
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus salivarius and application thereof. The lactobacillus salivarius is named as lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.18214 in 2019 and 07/12. The lactobacillus salivarius is applied to the preparation of medicines with the effects of relaxing bowels and/or reducing cholesterol. The lactobacillus salivarius can inhibit the growth and reproduction of harmful bacteria by metabolizing to generate lactic acid, promote intestinal peristalsis, increase the water content of excrement, and prevent and relieve constipation; the lactobacillus salivarius also has the function of reducing cholesterol, and when the lactobacillus salivarius reaches the intestinal tract after being ingested, the lactobacillus salivarius can reduce the absorption of cholesterol by the intestinal tract and convert part of cholesterol into cholate to be discharged out of the body.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus salivarius with a bowel relaxing effect, a cholesterol reducing effect, a strong acid resistance, a strong bile salt resistance and a strong gastrointestinal fluid resistance and application thereof.
Background
Along with the improvement of the living standard of people, the living rhythm is accelerated, and people with constipation and people with high cholesterol are more and more due to the reasons that people are biased to high-calorie fast food, lack of exercise, high pressure and the like. Constipation can make skin dull and dull, listlessness, dysphoria, abdominal distending pain, long-term constipation can also cause anorectal complications, and serious cases can even cause rectal cancer and colon cancer. High cholesterol damages blood vessels, causes cerebrovascular diseases and cardiovascular diseases, etc.
At present, according to the causes of constipation, the treatment schemes include general life treatment, drug treatment, biofeedback training and operation treatment. The most commonly used drug treatment scheme is divided into an external preparation and an internal preparation, and enema is a commonly used external purgative, which can play a role in short-term symptom relief, and symptoms still exist or even aggravate after the use, and the symptoms and the root causes are treated. The oral preparation generally achieves the aim of defecation by changing the permeability of intestinal tracts or using some pungent plant laxatives such as rheum officinale, senna leaf and the like, but has side effects of purgation, is easy to cause smooth muscle atrophy and the like, and aggravates constipation.
Drug therapy is also the option for most people, except by adjusting diet and increasing exercise amount in high cholesterol patients. However, it has been shown that chronic administration of statins for the treatment of high cholesterol causes non-specific muscle or joint pain and decreases side effects such as coenzyme Q10, an intermediate in the cholesterol synthesis pathway.
Probiotics are beneficial active microorganisms which can colonize human intestinal tracts, maintain micro-ecological balance in the human intestinal tracts and are beneficial to human health. Lactobacillus salivarius (Lactobacillus salivarius) is a probiotic listed in the list of strains for food in 2010, is gram positive, and has the effects of regulating immunity, resisting bacteria, reducing cholesterol and the like. However, the strains are different, the functional difference is larger, and the strain has different tolerance to gastrointestinal environment and intestinal functions, so that the obtained strain has strong acid resistance, strong bile salt resistance and strong gastrointestinal fluid resistance, reaches human intestinal tract with higher activity and can play a role in relaxing the bowels.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention aims to provide lactobacillus salivarius with strong acid resistance, strong bile salt resistance, strong gastrointestinal fluid resistance and the functions of relaxing the bowels and reducing cholesterol and application thereof, in particular to application in pharmaceutical compositions.
The strain is named as lactobacillus salivarius (Lactobacillus salivarius) HCS20-004, and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.18214 in the month 07 and 12 of 2019.
The application of the lactobacillus salivarius in preparing the medicine with the effect of reducing cholesterol.
The application of the lactobacillus salivarius in preparing the medicine with the effect of relaxing the bowels.
The application of the lactobacillus salivarius in preparing the medicine with the effects of reducing cholesterol and relaxing bowels.
The lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 is separated from the excreta of healthy 8-year-old girls, is a lactobacillus salivarius with strong capability of resisting the reverse environment of the digestive tract, has typical characteristics of lactobacillus, is gram-positive, and has circular colony, milky white, smooth surface, bulges in the middle and neat edges; the cells are in the shape of stubby bars. The physical and chemical characteristics are as follows: the cell morphology and physicochemical experimental results of the lactobacillus salivarius CGMCC No.18214 are shown in Table 1.
TABLE 1 cell morphology and physicochemical experiment results of Lactobacillus salivarius CGMCC No.18214
The 16S rDNA sequence of the Lactobacillus salivarius CGMCC No.18214 of the present invention was compared with the sequence similarity of the GenBank submitting strain by referring to the International related gene library in the American bioengineering information center (National Center for Biotechnology Information, NCBI), and the strain HCS20-004 was identified as Lactobacillus salivarius (Lactobacillus salivarius), and the comparison results are shown in Table 2.
TABLE 2 similarity of 16S rDNA sequence of Lactobacillus salivarius CGMCC No.18214 to GenBank submitted strain sequence
The lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 has the following beneficial effects when applied to a pharmaceutical composition:
1. the lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 is separated from human feces, so that the safety is high in application;
2. the lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 has excellent acid resistance and cholate resistance. When taken into the human body from the mouth, the composition can withstand the gastric environment with lower pH value and the high bile salt environment to reach the intestinal tract with higher survival rate;
3. the lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 has a good bowel relaxing function. Inhibiting growth and reproduction of harmful bacteria by metabolism to produce lactic acid, promoting intestinal peristalsis, increasing water content of feces, and preventing and relieving constipation;
4. the lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 has the function of reducing cholesterol. When the food reaches the intestinal tract after intake, the intestinal tract can reduce the absorption of cholesterol, and can convert part of cholesterol into cholate to be discharged outside the body.
Drawings
FIG. 1 is a cholesterol concentration standard curve.
The lactobacillus salivarius disclosed by the invention is named as lactobacillus salivarius (Lactobacillus salivarius) HCS20-004, and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.18214 and the preservation address of: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
Detailed Description
EXAMPLE 1 screening of Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 CGMCC No.18214 Strain
Adding 500uL of buffer protein lyophilized solution mixed with faeces sample into 5mL of buffer protein lyophilized solution, and diluting to 10 times by 10 times dilution method -4 Uniformly coating the diluted sample liquid on an acidification bile salt MRS culture medium plate, and carrying out anaerobic culture for 24-48 h at 37 ℃. Selecting single bacterial colony with typical characteristics (observation form, size, color, transparency and the like) of target bacterial strain, larger bacterial colony and stronger activity, carrying out streak purification culture on an MRS improved culture medium, and repeating the steps for 2-3 times until the bacterial colony characteristics in a streak plate are consistent; more than 2 individual colonies per purified plate were picked for smear, gram stain, and observed under a microscope for consistency in color, bacterial shape to determine if the colonies in the plate were pure cultures. If the observed results are consistent, taking the obtained pure culture (plate colony) as a suspected strain, numbering the corresponding plate, and identifying; if the observed results under the mirror are inconsistent, the operation is continued.
The MRS modified culture medium is prepared by: 10g/L of peptone, 3g/L of beef powder, 4g/L of yeast powder, 2g/L of dipotassium hydrogen phosphate, 2g/L of citric acid, 5g/L of sodium acetate, 20g/L of glucose, 0.58g/L of magnesium sulfate heptahydrate, 0.25g/L of manganese sulfate tetrahydrate, 0.6g/L of tween-80, 10g/L of calcium carbonate, 0.05g of neutral red (1% concentration of 5 mL), 10mL/L of tomato juice and 18g/L of agar powder, and adjusting the pH to 5.8; sterilizing at 115 deg.C for 30min.
The preparation method of the acidified bile salt MRS culture medium comprises the following steps: 10g/L of peptone, 3g/L of beef powder, 4g/L of yeast powder, 2g/L of dipotassium hydrogen phosphate, 2g/L of citric acid triammonium, 5g/L of sodium acetate, 20g/L of glucose, 0.58g/L of magnesium sulfate heptahydrate, 0.25g/L of manganese sulfate tetrahydrate, 0.6g/L of tween-80, 10g/L of calcium carbonate, 0.05g (1% concentration of 5 mL) of neutral red, 10mL/L of tomato juice, 0.1g/L of bovine bile salt and 18g/L of agar powder, and adjusting the pH to 5.5; sterilizing at 115 deg.C for 30min.
EXAMPLE 2 identification of Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 CGMCC No.18214 Strain
The pure culture isolated and purified in example 1 was further determined to be pure culture by streaking and smear microscopy, and then subjected to strain identification, including gram staining test, contact enzyme test and 16S rDNA full sequence sequencing identification. The final identification and separation of the strain is a strain of lactobacillus salivarius, named lactobacillus salivarius (Lactobacillus salivarius) HCS20-004, which is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.18214 in 2019, 07 and 12.
The lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 strain is gram positive, the colony is round, milky white, smooth in surface, convex in the middle and neat in edge; the cells are in the shape of stubby bars. The physical and chemical characteristics are as follows: as the contact enzyme-negative and oxidase-negative, D-galactose, D-glucose, D-fructose, D-mannose, L-rhamnose, mannitol, sorbitol, N-acetyl-glucosamine, maltose, melibiose, sucrose, raffinose, xylitol, and D-arabitol can be used.
EXAMPLE 3 Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 Strain culture experiments
The method comprises the following steps:
1. primary culture: taking out the strain freezing tube preserved at-80 ℃, thawing, inoculating 1mL (one freezing tube) of bacterial suspension into 10mL of modified lactobacillus culture medium, and standing and culturing at 37 ℃ for 17 hours;
2. dipping a trace of residual liquid in the freezing tube by using an inoculating loop, streaking on a flat plate, culturing for 24-48 h at 37 ℃, and observing colonies; a small amount of bacterial colony smear is picked, stained and the bacterial body under a microscope is observed;
3. secondary culture: transferring the bacterial suspension obtained by primary culture into 100mL of improved lactobacillus culture medium according to the inoculum size of 5%, and standing and culturing for 17 hours at 37 ℃;
4. and (3) three-stage culture: transferring the bacterial suspension obtained by the secondary culture into 300mL of improved lactobacillus culture medium according to the inoculum size of 5 percent, and standing and culturing for 17 hours at 37 ℃;
5. detecting the pH value and OD value of each level of culture solution, taking about 5mL of three-level culture solution by using a 10mL centrifuge tube to count the viable count, centrifuging the three-level culture solution (12000 r/min,10 min), discarding the supernatant, wiping the supernatant remained on the wall of the centrifuge tube, collecting bacterial mud, and calculating the bacterial mud yield;
6. after bacterial mud is collected by the three-stage culture solution, a micro bacterial mud is dipped by an inoculating loop and is coated on a glass slide, and bacterial bodies are observed under a microscope in a dyeing way.
The formula of the improved lactobacillus culture medium comprises the following components: 10g/L of yeast peptone, 3g/L of beef powder, 4g/L of yeast powder, 2g/L of monopotassium phosphate, 2g/L of citric acid, 5g/L of sodium acetate, 20g/L of glucose, 0.58g/L of magnesium sulfate heptahydrate, 0.25g/L of manganese sulfate tetrahydrate, 0.6g/L of tween-80, 10mL/L of tomato juice, regulating the pH to 6.5-7.0, subpackaging, and sterilizing at 115 ℃ for 30min.
TABLE 3HCS20-004 strain culture experimental data sheet
The observed colony forms are consistent, the single colony is about 2mm, round, milky white and smooth in surface. Gram-positive staining, rod-shaped, V-shaped, convex at both ends, convex in the middle, or shuttle-shaped, and the like, and is occasionally bifurcated and free of spores.
EXAMPLE 4 gastrointestinal adverse environmental test of Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 CGMCC No.18214 Strain
Lactobacillus salivarius (Lactobacillus salivarius) HCS 20-004's ability to withstand the reverse environment of the gut is a prerequisite for its ability to reach and survive, colonize and function in the gut with high survival rates.
1. Acid resistance test
The bacterial solutions of the three passages are respectively inoculated into a blank control culture medium and a basic MRS culture medium with the pH value of 3.0 according to the inoculation amount of 10 percent. And (3) carrying out stationary culture at 37 ℃ for 17 hours, sampling, carrying out 10-time serial dilution with sterilized normal saline, respectively taking 1000 mu L of bacterial liquid with proper dilution, carrying out mixed bacteria counting operation, repeating each dilution for 2 times, and carrying out stationary culture at 37 ℃ for 36-48 hours and counting.
Acid resistance test data index:
the number of viable bacteria (expressed as N') measured in the medium at different pH conditions; viable count measured in a blank control test (N 0 Expressed) and the acid-fast viable bacteria number logarithmic ratio is calculated as follows:
acid-resistant survival (%) = lg cfu N'/lg cfu N of the test strain 0 ×100％；
TABLE 4HCS20-004 acid resistance test data sheet
As shown in Table 4, the survival rate of HCS20-004 strain after 17 hours at pH3.0 is 90.0%, which indicates that the strain can still maintain higher activity after inhibition by gastric acid, thereby exerting its probiotic effect.
2. Test for bile salt resistance
Bacterial solutions of the three passages were inoculated into modified lactic acid bacteria culture media containing no ox gall salt (blank control) and 0.3%, 0.5%, 1.0% and 1.5% concentration of ox gall salt (sigma) according to an inoculum size of 10%, and were subjected to stationary culture at 37℃and sampling for 17 hours, and the number of viable bacteria was measured.
Data index of bile salt resistance test:
n for measuring viable count of blank control 0 The number of viable bacteria measured under other bile salt concentration conditions is represented by N', and the logarithmic ratio of the number of viable bacteria resistant to bile salts is represented byThe calculation formula is as follows:
test strain bile salt tolerance test survival (%) = lgcfu N "/lgcfu N 0 ×100％；
TABLE 5HCS20-004 bile salt resistance test data sheet
As shown in Table 5, although the HCS20-004 strain gradually decreased in viable count with increasing bile salt concentration, the survival rate was as high as 90.71% under the 1.5% bile salt concentration treatment condition.
3. Simulated gastric fluid test
Shaking up bacterial liquid after three passages, taking 10mL of bacterial suspension, centrifuging (5000 Xg, 10min,4 ℃) to obtain bacterial mud, flushing with PBS buffer solution for 2 times, re-suspending the obtained bacterial mud in 10mL of simulated gastric fluid, digesting for 3h at 37 ℃, and sampling and measuring the number of living bacteria respectively at 0h and 3 h.
The number of viable bacteria in the third-generation culture solution of the test strain after the third generation of activation is represented by N, and the number of viable bacteria measured by counting after digestion for 3 hours in the simulated gastric fluid culture medium is represented by N # The survival rate of the simulated gastric juice test of the tested strain is expressed as follows:
survival rate (%) =lgcfu N of simulated gastric juice test of the tested strain # /lgcfuN×100％；
TABLE 6HCS20-004 simulated gastric fluid test data sheet
As shown in Table 6, the HCS20-004 strain has strong survival ability in simulated gastric fluid, the survival rate can still reach 96.6% after 3 hours treatment, and the activity can still be kept high after the HCS20-004 strain stays in the stomach for a long time.
4. Simulated intestinal juice test
Shaking and shaking bacterial liquid for three times, taking 10mL of bacterial suspension, centrifuging (5000 Xg, 10min,4 ℃) to obtain bacterial mud, flushing with PBS buffer solution for 2 times, re-suspending the obtained bacterial mud in 10mL of artificial intestinal liquid, culturing at 37 ℃, and sampling and measuring the number of living bacteria respectively at 0h, 2h and 4 h.
The number of viable bacteria in the third generation culture solution of the test strain after the third generation is activated is represented by N, the number of viable bacteria counted after 2 hours and 4 hours of culture in the simulated intestinal fluid culture solution is represented by N, and the calculation formula of the survival rate of the simulated intestinal fluid test of the test strain is as follows:
survival rate (%) = lgcfun×lgcfun×100% of simulated intestinal fluid test of the tested strain;
TABLE 7HCS20-004 simulated intestinal juice test data sheet
As shown in Table 7, the HCS20-004 strain has strong survival ability in simulated intestinal fluid, and the survival rate of the strain is still as high as 98.9% when the simulated intestinal fluid is treated for 4 hours.
In conclusion, the strain has strong acid resistance and bile salt resistance, and can effectively resist the influence of gastrointestinal fluid, so that the strain can still maintain high activity after passing through the alimentary canal.
EXAMPLE 5 laxative efficacy test of Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 CGMCC No.18214 Strain
The test operation steps are as follows:
1. 40 clean-grade Kunming (KM) female mice were selected and weighed 18-22g and divided into four groups of 10 mice each.
2. The indoor temperature is controlled to be 22+/-1.5 ℃, the humidity is controlled to be 50+/-10%, the working illuminance is 160-280lx, and the noise is less than 60dB. The model of constipation in mice was established by oral gavage administration of the modeling drug loperamide hydrochloride (0.15 mg/mL).
3. 2 test dose groups, one solvent control group (distilled water) and one model control group were set. Test dose groups mice were given once a day with 4g/kg and 2g/kg of Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 lyophilized powder; the solvent control group is irrigated once a day, and the volume of the irrigated stomach is 10ml/kg; the test lasts for 7 days, and the defecation condition of the animal is reflected by recording the number of black bowel movements and the first black bowel movement time in 6 hours of the test animal.
The test results are shown in Table 8.
TABLE 8 results of the laxative efficacy test of Lactobacillus salivarius (HCS 20-004)
As shown by the test results in Table 8, the number of the black discharge particles in the 6h period of the C group and the D group is higher than that in the A group and the B group, and the first black discharge time is shorter than that in the A group and the B group. Namely, under the test condition, the HCS20-004 freeze-dried powder is continuously administered for 7 days under the dosage of 2g/kg and 4g/kg, and has certain defecation promoting effect.
EXAMPLE 6 test of cholesterol-lowering efficacy of Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 CGMCC No.18214 Strain
1. Drawing of a Standard Curve
1.1 1mg/mL cholesterol solution preparation
0.1g cholesterol was fixed to 100mL with absolute ethanol, filtered through a 0.22 μm filter, sterilized and stored at 4℃until use.
1.2 preparation of Cholesterol Standard solution
The cholesterol solutions of 50. Mu.L, 100. Mu.L, 150. Mu.L, 200. Mu.L, 250. Mu.L and 300. Mu.L of 1mg/mL were respectively aspirated into 10mL clean flasks and the volumes were determined with absolute ethanol to prepare standard solutions of 5. Mu.g/mL, 10. Mu.g/mL, 15. Mu.g/mL, 20. Mu.g/mL, 25. Mu.g/mL and 30. Mu.g/mL.
1.3 Standard cholesterol Curve drawing
Sucking 4mL of cholesterol standard solution, blowing nitrogen in a 50mL centrifuge tube, adding 4mL of phthalic dicarboxaldehyde solution, shaking, standing for 10min, adding 2.0mL of concentrated sulfuric acid, mixing, standing for color developmentAnd (3) for 10min, performing blank zeroing by using phthalic dicarboxaldehyde and concentrated sulfuric acid without adding cholesterol standard solution, and measuring the absorbance at 550 nm. Cholesterol concentration (μg/mL) is plotted on the abscissa, and OD550 is plotted on the ordinate. The linear regression equation is calculated to be y=0.0167x+0.0171, and the correlation coefficient is calculated to be R 2 = 0.9953. The standard curve is shown in figure 1.
2. Preparation of test strains
(1) Primary culture: taking out the strain cryopreservation tube preserved at-80 ℃, thawing, uniformly mixing, streaking the 1-cycle bacteria liquid on an MRS solid flat plate, and standing and culturing at 37 ℃ for 36-48h;
(2) Secondary culture: selecting single colony to 5mL MRS liquid culture medium, and culturing at 37 ℃ for 20h;
(3) And (3) three-stage culture: inoculating 5mL of bacterial liquid into 100mL of MRS liquid culture medium, and culturing at 37 ℃ for 17h;
the MRS culture medium is prepared by: 10g/L of yeast peptone, 3g/L of beef powder, 4g/L of yeast powder, 2g/L of monopotassium phosphate, 2g/L of citric acid, 5g/L of sodium acetate, 20g/L of glucose, 0.58g/L of magnesium sulfate heptahydrate, 0.25g/L of manganese sulfate tetrahydrate, 0.6g/L of tween-80, 10mL/L of tomato juice and 20g/L of agar, and sterilizing for 30min by regulating the pH to 6.5 and 115 ℃.
3. Colorimetric cholesterol content determination by phthalic dicarboxaldehyde
And inoculating the bacterial liquid after three-stage culture into MRS-CHOL culture medium according to an inoculum size of 1%. The control was MRS-CHOL medium without inoculation. Culturing at 37deg.C for 24h, at 800r/min, centrifuging at 4deg.C for 10min, and collecting supernatant.
4mL of the centrifuged supernatant was taken, 2.0mL of 50% KOH solution was added thereto, and after mixing uniformly, 3.0mL of 95% ethanol was added thereto, followed by vortexing for 1min. Placing the centrifuge tube in a water tank at 60 ℃, standing for 15min, taking out, rapidly cooling to room temperature, adding 3.0mL of n-hexane into the centrifuge tube, adding 2.0mL of distilled water after uniform mixing, sealing and vortex oscillating for 1min, standing for 15min, taking 2.0mL of n-hexane layer solution into a clean centrifuge tube, adding 2.0mL of phthalic aldehyde (OPA) solution after volatilizing the solvent by a nitrogen blow dryer at 60 ℃, uniformly mixing, standing for 10min at room temperature, adding 1.0mL of concentrated sulfuric acid, vortex oscillating, developing for 10min, and measuring OD 550 According to standard curveThe line calculates the cholesterol content in the sample.
The cholesterol removal rate of the test strain was calculated as follows:
wherein: c (C) 0 Measured concentration of cholesterol in the non-inoculated MRS-CHOL medium, μg/mL; c is the measured concentration of cholesterol in the supernatant of fermentation broth centrifugation obtained after inoculation of Lactobacillus salivarius (Lactobacillus salivarius) HCS20-004 culture, μg/mL.
The formula of the high cholesterol medium (MRS-CHOL) comprises the following components: MRS liquid culture medium, 2g/L sodium thioglycolate, 3g/L bovine bile salt (import), 115 ℃, sterilization for 30min, and 0.1g/L cholesterol solution.
TABLE 9 test results of efficacy of Lactobacillus salivarius (HCS 20-004) in lowering cholesterol
Note that: control strain H2001 is Lactobacillus salivarius.
According to the test results shown in Table 9, the control strain H2001 has no cholesterol degradation capability, while the lactobacillus salivarius (HCS 20-004) has a cholesterol degradation rate of 47.8% in an MRS culture medium, which indicates that the lactobacillus salivarius (HCS 20-004) has better cholesterol degradation capability.
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CGACTCTCTG TCCCTTAGAC GGCTGGCTCC TTGCGGTTAC CCCACCGGCT TTGGGTGTTA 60
CAAACTCTCA TGGTGTGACG GGCGGTGTGT ACAAGGCCCG GGAACGTATT CACCGCGACA 120
TGCTGATTCG CGATTACTAG CGATTCCGAC TTCATGTAGG CGAGTTGCAG CCTACAATCC 180
GAACTGAGAA CGGCTTTAAG AGATTAGCTA AACCTCGCGG TCTTGCGACT CGTTGTACCG 240
TCCATTGTAG CACGTGTGTA GCCCAGGTCA TAAGGGGCAT GATGACTTGA CGTCGTCCCC 300
ACCTTCCTCC GGTTTGTCAC CGGCAGTCTC GCCAGAGTGC CCAACTTAAT GCTGGCAACT 360
GACAACAAGG GTTGCGCTCG TTGCGGGACT TAACCCAACA TCTCACGACA CGAGCTGACG 420
ACAGCCATGC ACCACCTGTC ACTTTGTCCC CGAAGGGAAA GCCTAATCTC TTAGGTGGTC 480
AAAGGATGTC AAGACCTGGT AAGGTTCTTC GCGTTGCTTC GAATTAAACC ACATGCTCCA 540
CCGCTTGTGC GGGCCCCCGT CAATTCCTTT GAGTTTCAAC CTTGCGGTCG TACTCCCCAG 600
GCGGAATGCT TATTGCGTTA GCTGCGGCAC TGAAGGGCGG AAACCCTCCA ACACCTAGCA 660
TTCATCGTTT ACGGCGTGGA CTACCAGGGT ATCTAATCCT GTTTGCTACC CACGCTTTCG 720
AACCTCAGCG TCAGTTACAG ACCAGAGAGC CGCTTTCGCC ACTGGTGTTC TTCCATATAT 780
CTACGCATTT CACCGCTACA CATGGAGTTC CACTCTCCTC TTCTGCACTC AAGTCTTCCA 840
GTTTCCAATG CACTACTCCG GTTAAGCCGA AGGCTTTCAC ATCAGACTTA AAAGACCGCC 900
TGCGTTCCCT TTACGCCCAA TAAATCCGGA CAACGCTTGC CACCTACGTA TTACCGCGGC 960
TGCTGGCACG TAGTTAGCCG TGACTTGCTG GTTAGATACC GTCATCGAAT GAACAGTTAC 1020
TCTCACTCGT GTTCTTCTCT AACAACAGAG TTTTACGATC CGAAGACCTT CTTCACTCAC 1080
GCGGCGTTGC TCCATCAGAC TTGCGTCCAT TGTGGAAGAT TCCCTACTGC TGCCTCCCGT 1140
AGGAGTTTGG GCCGTGTCTC AGTCCCAATG TGGCCGATCA ACCTCTCAGT TCGGCTACGT 1200
ATCATCACCT TGGTAGGCCG TTACCCCACC AACTAGTTAA TACGCCGCGG GTCCATCTAA 1260
AAGCGATAGC AGAACCATCT TTCATCTAAG GATCATGCGA TCCTTAGAGA TATACGGTAT 1320
TAGCACCTGT TTCCAAGTGT TATCCCCTTC TTTTAGGCAG GTTACCCACG TGTTACTCAC 1380
CCGTCCGCCA CTCAACTTCT TACGGTGAAT GCAAGCATTC GGTGTAAGAA AGTTTCGTTC 1440
GACTTGCATC ATAGACACG 1459
Claims (4)
1. A strain of Lactobacillus salivarius, characterized in that the strain is named Lactobacillus salivarius (Lactobacillus salivarius)
salivarius) HCS20-004, which has been deposited in the China general microbiological culture Collection center, with a collection number of CGMCC No.18214, at 2019, month 07, and day 12.
2. Use of a strain of lactobacillus salivarius as claimed in claim 1 in the manufacture of a medicament for use in reducing cholesterol.
3. Use of a strain of lactobacillus salivarius as claimed in claim 1 in the manufacture of a medicament for use in the treatment of constipation.
4. Use of a strain of lactobacillus salivarius as claimed in claim 1 in the manufacture of a medicament for use in the reduction of cholesterol and in the relief of constipation.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113337440B (en) * | 2021-06-21 | 2023-02-21 | 美益添生物医药(武汉)有限公司 | Lactobacillus salivarius MG-587 and application thereof |
| CN115927045B (en) * | 2022-07-13 | 2023-10-20 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Lactobacillus salivarius 069 with cholesterol reducing and liver injury relieving functions caused by hyperlipidemia and application thereof |
| CN115969888A (en) * | 2022-10-25 | 2023-04-18 | 深圳未知君生物科技有限公司 | Saliva combined lactobacillus and application thereof in preparation of medicine for treating cancer |
| CN117384788B (en) * | 2023-10-10 | 2024-03-22 | 广东悦创生物科技有限公司 | Saliva combined lactobacillus SM4 and application thereof in preparation of whitening and cholesterol lowering foods and medicines |
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| CN103897998A (en) * | 2012-12-25 | 2014-07-02 | 北京中天神舟航天食品技术研究院 | Lactobacillus salivarius, applications thereof, functional food compositions and preparation method of the functional food composition |
| EP3412766A1 (en) * | 2017-06-06 | 2018-12-12 | Casen Recordati, S.L. | Lactobacillus salivarius strain, composition comprising the same, and uses thereof |
| WO2019169181A1 (en) * | 2018-02-28 | 2019-09-06 | Evelo Biosciences, Inc. | Compositions and methods for treating cancer using lactobacillus salivarius |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002016554A1 (en) * | 2000-08-25 | 2002-02-28 | Wakamoto Pharmaceutical Co.,Ltd. | Probiotics products containing lactic acid bacterium |
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| CN103897998A (en) * | 2012-12-25 | 2014-07-02 | 北京中天神舟航天食品技术研究院 | Lactobacillus salivarius, applications thereof, functional food compositions and preparation method of the functional food composition |
| EP3412766A1 (en) * | 2017-06-06 | 2018-12-12 | Casen Recordati, S.L. | Lactobacillus salivarius strain, composition comprising the same, and uses thereof |
| WO2019169181A1 (en) * | 2018-02-28 | 2019-09-06 | Evelo Biosciences, Inc. | Compositions and methods for treating cancer using lactobacillus salivarius |
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| Effects of a Lactobacillus salivarius mixture on performance, intestinal health and serum lipids of broiler chickens;Parisa Shokryazdan等;《PLoS ONE》;第12卷(第5期);e0175959 * |
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