CN114990011A - Lactobacillus reuteri capable of reducing cholesterol and inhibiting gardnerella and application - Google Patents
Lactobacillus reuteri capable of reducing cholesterol and inhibiting gardnerella and application Download PDFInfo
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- CN114990011A CN114990011A CN202210572722.9A CN202210572722A CN114990011A CN 114990011 A CN114990011 A CN 114990011A CN 202210572722 A CN202210572722 A CN 202210572722A CN 114990011 A CN114990011 A CN 114990011A
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- lactobacillus reuteri
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- hcs02
- cholesterol
- inhibiting
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus reuteri with cholesterol-reducing and gardnerella vaginalis-inhibiting functions and application thereof. The Lactobacillus reuteri is named as Lactobacillus reuteri HCS02-001, and the strain is preserved in China general microbiological culture Collection center (CGMCC) at 27 days 04/2020 with the preservation number of CGMCC No. 19746. The Lactobacillus reuteri HCS02-001 has the functions of reducing cholesterol and inhibiting gardnerella vaginalis.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus reuteri capable of reducing cholesterol and inhibiting gardnerella and application thereof.
Background
In the vagina of a healthy woman, a complex microbial system is formed, and various bacteria including probiotics and pathogenic bacteria in a balanced state are contained in the vagina, so that the vagina health is influenced by the combined action of the probiotics and the pathogenic bacteria. Gardnerella vaginalis is a main pathogenic bacterium causing gynecological diseases, and under the influence of certain factors, such as antibiotic use and the like, the gardnerella vaginalis overgrows to cause vaginitis and other diseases. The existing treatment mainly aims to improve the symptoms of gynecological diseases, adjust vaginal microbial flora and restore vaginal microbial balance so as to resist the invasion of external pathogenic bacteria. Therefore, the development of the bacterial strain with strong inhibiting effect on gardnerella vaginalis is particularly important, and the bacterial strain has acid resistance, and has no stimulation or harm to the interior of the vagina.
High cholesterol levels can cause cardiovascular disease, atherosclerosis, and high cholesterol levels are associated with many metabolic syndromes, resulting in the development of obesity, hypertension, and hyperlipidemia. At present, cholesterol is reduced mainly through drug treatment, which causes muscle spasm of organisms and has large side effect. Some patients select diet therapy, the effect of the medicine is reduced after long-term taking, the long-term taking is difficult to persist, the probiotic preparation with the function of reducing the cholesterol is produced at the same time, and the cholesterol degradation rate of some probiotics is generally low at present, and the formula is listed in table 1.
Lactobacillus reuteri is widely present in the intestinal tracts of humans and animals, and is abundant in source. The product has various probiotic effects, can regulate intestinal flora, effectively prevent diarrhea, inhibit reproduction of pathogenic microorganisms, and reduce intestinal diseases. The lactobacillus reuteri has more probiotic functions, one of the probiotic functions is participated in cholesterol metabolism, enters the intestinal tract through a digestive system after being taken, has the characteristics of acid resistance and bile salt resistance, and plays a probiotic role on the organism. The lactobacillus reuteri has the functions of reducing cholesterol and inhibiting gardnerella vaginalis, and at present, a plurality of lactic acid bacteria which are separated out through research have the functions of inhibiting gardnerella vaginalis, and vaginal diseases are eliminated by externally adjusting vaginal flora balance, so that the development and development of a cholesterol-reducing and gardnerella vaginalis-inhibiting microecological preparation have important significance for improving the health of people.
TABLE 1 Cholesterol degradation Rate for the bacterial preparations published in the prior art
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a lactobacillus reuteri strain which has strong acid resistance, strong bile salt resistance, strong gastrointestinal fluid resistance, a function of reducing cholesterol and a function of inhibiting gardnerella vaginalis and can be adhered to the vagina and an application thereof, in particular to an application in food or pharmaceutical compositions.
The Lactobacillus reuteri is named as Lactobacillus reuteri HCS02-001, and is preserved in China general microbiological culture Collection center (CGMCC) at 27 days 04 in 2020 and has the preservation number of CGMCC No. 19746.
The lactobacillus reuteri strain is applied to preparation of a medicine for reducing cholesterol.
The lactobacillus reuteri strain is applied to preparation of medicines for inhibiting gardnerella vaginalis.
A pharmaceutical preparation with cholesterol lowering and/or gardnerella vaginalis inhibiting functions comprises an effective dose of lactobacillus reuteri and pharmaceutically acceptable auxiliary materials.
A microbial inoculum with cholesterol lowering and/or vaginal Gardner bacteria inhibiting functions is a microbial inoculum containing the lactobacillus reuteri.
The present invention provides a composition of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 strain or a culture solution thereof.
The present invention provides a food composition of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 strain or a culture solution thereof.
The present invention provides a pharmaceutical composition of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 strain or a culture solution thereof.
The food composition can be a health food composition or a fermented food composition.
The formulation of the food composition or the pharmaceutical composition is not limited, and for example, it may be formulated into powder, tablet, pill, capsule, granule, oral liquid, etc.
The cholesterol lowering function can be used in food or pharmaceutical compositions, and the inhibition of gardnerella vaginalis can be used in pharmaceutical compositions.
The lactobacillus reuteri HCS02-001 is separated from milk curd in a landau in reobelch Manchurian, inner Mongolia, is a lactobacillus reuteri strain with the functions of reducing cholesterol and inhibiting gardnerella vaginalis, has the typical characteristics of the lactobacillus, is gram-positive, has no spores, and has rod-shaped cells which are arranged in pairs. The physical and chemical characteristics are as follows: the contact enzyme is negative, the oxidase is negative, and the cell morphology and the physicochemical experiment results of the lactobacillus reuteri CGMCC No.19746 are detailed in the table 2.
TABLE 2 cell morphology and physicochemical experiment results of Lactobacillus reuteri CGMCC No.19746
The 16S rDNA sequence of Lactobacillus reuteri CGMCC No.19746 of the present invention was compared with the sequence similarity of the GenBank-submitted strain by consulting the International Union of genes related in the National Center for Biotechnology Information, NCBI, to identify the strain HCS02-001 as Lactobacillus reuteri (Lactobacillus reuteri), and the results of the comparison are shown in Table 3.
TABLE 3 comparison of similarity of 16S rDNA sequence of Lactobacillus reuteri CGMCC No.19746 and GenBank submitted strain sequence
Compared with the prior art, the invention has the following beneficial effects:
1. the Lactobacillus reuteri HCS02-001 has the functions of reducing cholesterol and inhibiting Gardnerella vaginalis.
2. According to the Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001, the cholesterol degradation rate of the cholesterol-lowering function reaches 51.5%.
3. The Lactobacillus reuteri HCS02-001 has the function of inhibiting Gardnerella vaginalis, and 3 times of concentration of fermentation liquor and fermentation supernatant fluid has an inhibition zone with the diameter of more than 14.6mm, so that the inhibition capability is strong;
4. the Lactobacillus reuteri HCS02-001 has better vaginal cell adhesion capability and can adhere to the vagina to play a role in inhibiting bacteria.
5. The Lactobacillus reuteri HCS02-001 has excellent acid resistance and cholate resistance. When ingested orally into the human body, it is able to withstand the lower pH gastric environment and the high bile salt environment to reach the intestinal tract with a higher survival rate.
6. The Lactobacillus reuteri HCS02-001 is separated from food raw material milk tofu, the components and raw materials adopted by a culture medium and a neutralizer in the strain purification and fermentation process are food grade, and the Lactobacillus reuteri HCS02-001 is verified to be safe and can be used for food fermentation, freeze-dried strain powder, food or medicine and the like.
Drawings
FIG. 1a shows the result of 3 times concentration inhibition of L.reuteri HCS02-001 fermentation liquid on Gardner's bacteria.
FIG. 1b shows the results of 3-fold concentration inhibition of L.reuteri HCS02-001 supernatant in Gardner bacteria.
FIG. 2 is a standard curve for cholesterol.
The Lactobacillus reuteri with the functions of reducing cholesterol and inhibiting gardnerella vaginalis is named as Lactobacillus reuteri HCS02-001, and is preserved in China general microbiological culture Collection center (CGMCC No. 19746) at 27.04.2020 and has the preservation address as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
Detailed Description
Example 1 screening of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 CGMCC No.19746 Strain
1. Preliminary screening
(1) Sample enrichment: adding a small spoon of sample into sterilized buffer peptone water culture medium, mixing, and culturing in a 37 deg.C incubator for 24 h;
(2) adding 500 μ L into 4.5mL modified MRS after enrichment, mixing well, sequentially performing ten times of gradient dilution to obtain 10 -2 -10 -6 Diluting, then sucking 100 μ L of each dilution, adding to the surface of modified MRS solid plate, coating with coating rod, and adding 10 μ L of each dilution -1 And (4) scribing the dilution zones, making two plates for each dilution, inversely placing the plates into a self-sealing bag, and respectively placing the self-sealing bag into an incubator at 37 ℃ and an anaerobic bag for culturing for 48 hours.
2. Purification of
And selecting a single colony with typical characteristics of a target strain, a large colony and high activity, performing streak purification culture on an improved MRS solid culture medium, culturing for 24-48h at 37 ℃, and repeating the steps for 2-3 times until the characteristics of the colony in a streak plate are consistent.
3 microscopic examination
And picking more than 2 single colonies from each purified plate, smearing, gram staining, and observing whether the color and the thallus shape are consistent under a microscope, thereby determining whether the colonies in the plate are pure cultures.
The improved MRS culture medium comprises: 10g/L of peptone, 3g/L of beef powder, 4g/L of yeast powder, 2g/L of dipotassium 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, 800.6 g/L of Tween-A, 10g/L of calcium carbonate, 0.05g (1% concentration of 5mL) of neutral red, 10mL/L (v/v) of tomato juice and 20g/L of agar powder, and adjusting the pH to 5.5; sterilizing at 115 deg.C for 30 min.
Example 2 identification of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 CGMCC No.19746 Strain
The pure culture isolated and purified in example 1 was subjected to strain identification including gram stain test, catalase test and 16S rDNA full sequence sequencing identification. The finally identified and separated strain is Lactobacillus reuteri, is named as Lactobacillus reuteri HCS02-001, is preserved in China general microbiological culture Collection center (CGMCC) at 27 days 04 and 2020, and has the preservation number of CGMCC No. 19746.
The Lactobacillus reuteri HCS02-001 strain is gram-positive, and the cells are in the shape of polymorphous rods and are arranged in pairs. The physical and chemical characteristics are as follows: the catalase is negative, the oxidase is negative, and L-arabinose, D-galactose, D-glucose, D-fructose, maltose, lactose, melibiose, sucrose, and raffinose can be used.
Example 3 stress test of resistance to digestive tract by Lactobacillus reuteri HCS02-001 CGMCC No.19746 Strain
The ability of Lactobacillus reuteri HCS02-001 to withstand the adverse environment of the digestive tract 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 liquid after three passages is respectively inoculated in a blank control culture medium, a basic MRS culture medium with pH2.0 and a basic MRS culture medium with pH3.0 according to the inoculum size of 10 percent. Performing static culture at 37 ℃, sampling for 17h, performing a series of 10-fold dilutions by using sterilized normal saline, performing viable bacteria counting operation on 1000 mu L of bacterial liquid with appropriate dilution respectively, repeating the steps for 2 times at each dilution, and counting after static culture at 37 ℃ for 36-48 h.
Acid resistance test data indexes:
viable count (denoted by N') measured for media at different pH conditions; viable count (in N) in the blank control 0 Expressed), the acid-resistant viable cell count logarithmic ratio calculation formula is as follows:
the acid-resistant survival rate (%) of the tested strain is lg cfu N'/lg cfu N 0 ×100％；
TABLE 4 acid resistance test data table for Lactobacillus reuteri HCS02-001
As can be seen from Table 4, the survival rate of the HCS02-001 strain still reaches 85.52% after 17 hours under the condition of pH2.0, which shows that the bacterium can still maintain higher activity after the inhibition of gastric acid, thereby exerting the probiotic effect.
2. Bile salt resistance test
Respectively inoculating the bacterial liquid after three passages in MRS liquid culture medium with different cholate concentrations of 0.3%, 0.5%, 1.0% and 1.5% (imported sigma cholate) according to the inoculation amount of 10%, standing and culturing at 37 ℃, sampling for 17h, and determining the viable count.
N for measuring viable count of blank control 0 The viable count measured under other conditions of the bile salt concentration is expressed by N', and the calculation formula of the bile salt resistant viable count logarithmic ratio is as follows:
the survival rate (%) of the tested strain in the bile salt resistance test is lgcfuN'/lgcfu N 0 ×100％；
TABLE 5 Lactobacillus reuteri HCS02-001 Table for bile salt resistance test data
As shown in Table 5, although the number of viable bacteria of the HCS02-001 strain gradually decreased with the increase of the concentration of bile salts, the survival rate was still as high as 90.99% under the condition of 1.5% treatment of the concentration of bile salts, and the strain had strong bile salt resistance.
3. Simulated gastric fluid test
Shaking the bacterial liquid after three passages evenly, centrifuging 10mL of bacterial suspension (5000 Xg, 10min, 5 ℃) to obtain bacterial sludge, washing the bacterial sludge for 2 times by using PBS buffer solution, suspending the obtained bacterial sludge in 10mL of simulated gastric juice, digesting the bacterial sludge for 3 hours at 37 ℃, and sampling to measure the number of viable bacteria.
The viable count of the test strain after the third generation of activation in the third generation culture medium is represented by N, and the viable count measured after digestion for 3 hours in the simulated gastric juice culture medium is represented by N # The survival rate of the tested strain simulated gastric juice test is represented by the following formula:
the survival rate (%) of the tested strain in simulated gastric fluid test is lgcfu N # /lgcfuN×100％；
TABLE 6 data sheet for simulated gastric fluid test of Lactobacillus reuteri HCS02-001
As can be seen from Table 6, the HCS02-001 strain has strong survival ability in simulated gastric juice, the survival rate is as high as 98.71% after 3h treatment, and the strain still maintains high activity for a long time in the stomach.
4. Simulated intestinal fluid test
Shaking the bacterial liquid after three passages evenly, centrifuging 10mL of bacterial suspension (5000 Xg, 10min, 5 ℃) to obtain bacterial sludge, washing the bacterial sludge for 2 times by using PBS buffer solution, suspending the obtained bacterial sludge in 10mL of artificial intestinal juice, culturing the bacterial sludge at 37 ℃, and sampling for 2h and 4h respectively to measure the number of viable bacteria.
The viable count of the test strain after the third generation of activation in the third generation culture solution is represented by N, the viable count measured after 2 hours and 4 hours of culture in the simulated intestinal juice culture solution is represented by N, and the survival rate calculation formula of the test strain simulated intestinal juice test is as follows:
the tested strain simulates intestinal fluid test survival rate (%) ═ lgcfuN/lgcfuN multiplied by 100%;
TABLE 7 data table of simulated intestinal fluid test of Lactobacillus reuteri HCS02-001
As can be seen from Table 7, the HCS02-001 strain has strong survival ability in simulated intestinal fluid, and when the simulated intestinal fluid is treated for 4 hours, the strain almost completely survives, and the survival rate is as high as 99.89%.
In conclusion, the strain has strong acid and bile salt resistance, and can effectively resist the influence of gastrointestinal fluids, so that high activity can be maintained after the strain passes through the digestive tract.
Example 4 Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 inhibition of Gardnerella vaginalis functional assay
1. Concentrating fermentation supernatant of Lactobacillus reuteri HCS02-001
Thawing a strain HCS02-001 frozen tube at room temperature, uniformly mixing, taking 1-ring frozen tube bacterial liquid, streaking on an MRS solid plate, performing static culture at 37 ℃ for 48h, selecting a single bacterial colony, culturing in 5mL of MRS liquid culture medium at 37 ℃ for 20h, performing amplification culture on 5% of inoculum size to 100mL of MRS liquid culture medium, and culturing at 37 ℃ for 24 h.
Fermentation liquor sample: and concentrating the cultured strain fermentation liquor by 3 times, and storing at 4 ℃ for later use.
Fermentation broth supernatant sample: centrifuging the cultured strain fermentation liquid at 4 deg.C and 4500g for 10min, collecting supernatant, concentrating 3 times, and storing at 4 deg.C.
A sterile medium was prepared as a blank, and the blank was similarly concentrated to 3-fold by heating and left at 4 ℃ for further use.
The MRS culture medium: 10g/L of yeast peptone, 3g/L of beef powder, 4g/L of yeast extract, 2g/L of potassium dihydrogen phosphate, 2g/L of citric acid, 5g/L of sodium acetate, 20g/L of glucose, 0.58g/L of magnesium sulfate, 0.25g/L of manganese sulfate, 800.6 g/L of tween, 10mL/L (v/v) of tomato juice and pH 6.5.
2. Preparation of Gardnerella vaginalis test Strain
Inoculating 1mL bacterial solution of Gardner vaginal bacteria in 100mL in TSB liquid medium added with sterile defibrinated sheep blood (addition amount is 50g/L), culturing at 37 deg.C for 24h (micro-anaerobic aerogenic bag culture (5% CO) 2 ))。
The TSB culture medium: 17g/L of tryptone, 3g/L of plant peptone, 5g/L of sodium chloride, 2.5g/L of dipotassium phosphate, 2.5g/L of glucose, 20g/L of solid agar powder and 7.3 +/-0.2 of pH.
3. Preparation of indicator-containing double-layer plate
15-20mL of TSB medium supplemented with sterile defibered sheep blood (added in an amount of 50g/L) was poured onto a dish and the agar was allowed to solidify to room temperature. Placing 4 oxford cups on average, and collecting 100 μ L of Gardnerella vaginalis bacterial solution (concentration about 1 × 10) 8 CFU/mL) and 10mL of 1.5% agar medium (air-dried to be not hot) are mixed and poured into a plate to ensure that the concentration of the pathogenic bacteria liquid in the plate is about 1 × 10 6 CFU/mL, after the agar solidified, taking away the Oxford cup to form a sample hole with the diameter of 8 mm.
4. Adding antibacterial substance strain HCS02-001 fermentation liquid and fermentation supernatant
Adding 100 μ L of 3 times concentrated sample of strain fermentation liquid into 3 sample adding holes of the plate, adding 3 times concentrated solution of sterilized MRS culture medium into 1 other hole as blank control, carefully placing the plate in 37 deg.C incubator for culturing for 48h (micro anaerobic gas production package culture (5% CO) 2 ))。
5. Measuring the diameter of the zone of inhibition
After 48h of culture, the bacteriostatic circle is observed, and the diameter of the bacteriostatic circle is measured by a vernier caliper.
6. Test results for inhibiting Gardnerella vaginalis
TABLE 8 bacteriostatic spectra of Gardnerella vaginalis
As can be seen from the test results shown in Table 8 and FIG. 1, the 3-fold concentration of the fermentation liquid of the strain HCS02-001 and the 3-fold concentration of the fermentation supernatant both have inhibition zones, the diameters of the inhibition zones both reach more than 14.6mm, and the strain HCS02-001 has an obvious inhibition effect on Gardner vaginal bacteria.
7. Contrasting the bacteriostatic effect of other strains
TABLE 9 comparison of other strains
Example 5 Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 vaginal cell adhesion test
1. Test method
Adhesion of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 to vaginal epithelial cells: selecting 30 healthy women with age of 20-30 years, normal gynecological examination, and asexual life history within 3 days, scraping posterior segment mucosal epithelial cells in vagina, suspending in 10mL RPM11640, washing cells with vortex mixer, removing endogenous bacteria, and adjusting cell concentration to 1 × 10 5 one/mL. Using commercial strain as control group, adjusting final concentration of bacterial liquid to be detected to 2 × 10 7 CFU/mL, mixing with 1mL of vaginal epithelial cell suspension, shaking and culturing at 37 ℃ for 1h, centrifuging at 1000r/min for 5min, washing with PBS for 3 times, discarding supernatant containing cells which are not adhered, smearing precipitate, drying, fixing with methanol for 15min, gram staining, randomly counting 50 epithelial cells under an oil mirror, and calculating the adhesion index of each epithelial cell, wherein the adhesion index is adhesive bacteria number/cell number multiplied by 100%.
2. Test results
Table 10 comparison of the adhesion indices of the different strains to vaginal epithelial cells (%,
)
the results show that different strains have different adhesion degrees to vaginal epithelial cells when the bacterial concentration is 2X 10 7 CFU/mL, the difference between the adhesion indexes of different strains has statistical significance, and the adhesion index of the lactobacillus reuteri is obviously higher than that of the commercial strain, which shows thatThe lactobacillus reuteri can adhere to cells in the vagina and colonize the environment to play a role.
Example 6 Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 functional in vitro assay for cholesterol lowering
1. Test method
(1) Drawing of standard curve
1mg/mL cholesterol solution: 0.1g of cholesterol, the volume is adjusted to 100mL by absolute ethyl alcohol, the mixture is filtered and sterilized by a 0.22 mu m filter membrane, and the mixture is stored at 4 ℃ for standby.
Preparing a cholesterol standard solution: the cholesterol solution of 50. mu.L, 100. mu.L, 150. mu.L, 200. mu.L, 250. mu.L, 300. mu.L 1mg/mL was pipetted into a 10mL clean volumetric flask, and the volume was 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.
Standard curve for cholesterol: absorbing 4mL of cholesterol standard solution into a 50mL centrifuge tube, blowing nitrogen flow to blow a solvent by a nitrogen blow-drying instrument at 60 ℃, adding 4mL of o-phthalaldehyde solution, shaking, standing for 10min, adding 2mL of concentrated sulfuric acid, mixing uniformly, developing for 10min, zeroing by taking the o-phthalaldehyde and the concentrated sulfuric acid which are not added with the cholesterol standard solution as blanks, and measuring the light absorption value at 550 nm. A cholesterol standard curve was plotted with cholesterol concentration (. mu.g/mL) as the abscissa and. DELTA.OD 550 as the ordinate.
(2) Preparation of test strains
And (3) streaking the strain to separate a single colony, selecting the single colony, inoculating the single colony in 10mL of a suitable culture medium, covering the culture medium with tin foil at a suitable temperature for 20h, transferring the single colony to 200mL of the culture medium again, and continuously covering the tin foil at a suitable temperature for 17h for subsequent tests.
3. O-phthalaldehyde colorimetric method for measuring cholesterol content (OPA method)
Inoculating the cultured bacterial liquid into 50mL of high-cholesterol culture medium according to the inoculation amount of 1%; the control was high cholesterol medium without inoculation. Culturing at 39 deg.C for 24h, 8000r/min, centrifuging at 4 deg.C for 10min, collecting supernatant, and determining by OPA method.
And (3) adding 2mL of 50% KOH into 4mL of centrifuged supernatant, uniformly mixing, adding 3mL of 95% ethanol, and shaking for 1min by vortex. Placing the centrifuge tube in a 60 ℃ water tank, standing for 15min, taking out, rapidly cooling to room temperature, adding 3mL of n-hexane into the centrifuge tube, uniformly mixing, adding 2mL of distilled water, covering, performing vortex oscillation for 1min, standing for 15min, taking 2mL of n-hexane layer solution into a clean centrifuge tube, volatilizing the solvent in a 60 ℃ nitrogen blow-drying instrument, adding 2mL of phthalic dicarboxaldehyde (OPA) solution prepared on the same day, uniformly mixing, standing for 10min at room temperature, adding 1mL of concentrated sulfuric acid, performing vortex oscillation, developing for 10min, measuring delta OD550, and calculating the cholesterol content in the sample according to a standard curve.
2. Test result calculation method
The cholesterol content of the sample was determined according to a standard curve.
In the formula: c 0 The actually measured concentration of cholesterol in the centrifugal supernatant of the non-inoculated culture solution is mu g/mL; c is the actually measured concentration of cholesterol in the supernatant fluid of the inoculated fermentation liquor centrifugation, mu g/mL.
Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 showed a cholesterol removal rate of 51.5%.
Example 7 Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 safety verification
The pathogenicity of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 was detected according to the safety inspection and evaluation technical guide principle (2010 edition) for the strains of the raw materials of the health food, and the results are shown in tables 11-16. The result shows that the strain HCS02-001 is nonpathogenic and has safety.
TABLE 11 Effect of the culture of strain HCS02-001 on the body weight of male mice (intraperitoneal injection)
Table 11 the results illustrate: the initial body weight and the final body weight of the male mice were not significantly different (P > 0.05) compared between the culture group and its corresponding control group, i.e., the strain HCS02-001 culture had no effect on the body weight of the male mice.
TABLE 12 Effect of the culture of strain HCS02-001 on the body weight of female mice (intraperitoneal injection)
Table 12 the results illustrate: the initial body weight and the final body weight of the female mice have no significant difference (P > 0.05) compared between the culture group and the corresponding control group, namely, the culture of the strain HCS02-001 has no influence on the body weight of the female mice.
TABLE 13 acute toxicity of cultures of strain HCS02-001 on mice (intraperitoneal injection)
Table 13 the results show that: the strain HCS02-001 culture stock solution was intraperitoneally injected into mice at a dose of 0.2mL for 1 time, and 21 days were continuously observed, and no toxic reaction or death of the test mice was observed.
TABLE 14 Effect of the culture of Strain HCS02-001 on Male mouse body weight (by oral gavage)
Table 14 the results show that: the initial and final body weights of the male mice were not significantly different (P > 0.05) compared between the culture group and its corresponding control group, i.e., the strain HCS02-001 culture had no effect on the body weight of the male mice.
TABLE 15 Effect of the culture of Strain HCS02-001 on female mouse body weight (by oral gavage)
Table 15 the results show that: both the initial and final body weights of female mice were not significantly different (P > 0.05) compared between the culture group and its corresponding control group, i.e., the strain HCS02-001 culture had no effect on the body weight of male mice.
TABLE 16 acute toxic effects of the culture of strain HCS02-001 on mice (by oral gavage)
Table 16 the results show that: the HCS02-001 culture stock solution and the concentrated solution are subjected to oral gavage on a mouse with the dosage of 20.0mL/kg BW, the gavage is continuously carried out for 3 days, 21 days are observed, and the toxic reaction or death phenomenon of the tested mouse is observed.
Example 8 method for culturing Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001
1. Recovering the frozen strains: taking a strain freezing tube stored in a low-temperature refrigerator, immediately putting the strain freezing tube into a water bath kettle at 37 ℃ for strain recovery for 15-30 s until all solids in the freezing tube are melted;
2. first stage culture
Transferring 1mL of the recovered strain into 10mL of a basal medium, carrying out shaking culture at 37 ℃ and 100rpm for 16-20h, and storing in a refrigerator at 4 ℃ for later use.
3. Second stage culture
Transferring the bacterial suspension obtained by the first-stage culture into a 100mL basal medium by an inoculum size of 5%, carrying out shake culture at 37 ℃ and 100rpm for 16-20h, and determining the pH value of the bacterial suspension to be 4.40-4.60 and the yield to be 1.00-1.30.
4. Three-stage culture
Transferring the bacterial suspension obtained by secondary fermentation into 300mL of optimized culture medium with the inoculation amount of 5%, loading liquid by 40-60%, carrying out shake culture at 37 ℃ and 100rpm for 16-20h, and determining the pH value of the bacterial suspension to be 4.60-4.80 and the yield to be 1.20-1.50%.
The basic culture medium comprises: 15g/L of yeast peptone, 10g/L of yeast extract, 20g/L of white granulated sugar, 5g/L of sodium acetate, 2g/L of citric acid monohydrate, 2g/L of potassium dihydrogen phosphate, 0.5g/L of magnesium sulfate, 0.01g/L of manganese sulfate and pH 6.80.
The optimized culture medium comprises: 20g/L of yeast peptone, 15g/L of yeast extract, 30g/L of brown sugar, 5g/L of sodium acetate, 2g/L of citric acid monohydrate, 2g/L of potassium dihydrogen phosphate, 0.5g/L of magnesium sulfate, 0.01g/L of manganese sulfate and pH 6.80.
The viable count of the bacterial suspension obtained by three-stage culture of Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001 can reach (3.0-5.0) x 10 9 CFU/mL。
<110> Jiangxi Renren health industry Co., Ltd
<120> lactobacillus reuteri capable of reducing cholesterol and inhibiting gardnerella and application
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>1485
<212>DNA
<213> Lactobacillus reuteri (Lactobacillus reuteri) HCS02-001
<400>1
ATACATGCAA GTCGTACGCA CTGGCCCAAC TGATTGATGG TGCTTGCACC TGATTGACGA 60
TGGATCACCA GTGAGTGGCG GACGGGTGAG TAACACGTAG GTAACCTGCC CCGGAGCGGG 120
GGATAACATT TGGAAACAGA TGCTAATACC GCATAACAAC AAAAGCCACA TGGCTTTTGC 180
TTGAAAGATG GCTTTGGCTA TCACTCTGGG ATGGACCTGC GGTGCATTAG CTAGTTGGTA 240
AGGTAACGGC TTACCAAGGC GATGATGCAT AGCCGAGTTG AGAGACTGAT CGGCCACAAT 300
GGAACTGAGA CACGGTCCAT ACTCCTACGG GAGGCAGCAG TAGGGAATCT TCCACAATGG 360
GCGCAAGCCT GATGGAGCAA CACCGCGTGA GTGAAGAAGG GTTTCGGCTC GTAAAGCTCT 420
GTTGTTGGAG AAGAACGTGC GTGAGAGTAA CTGTTCACGC AGTGACGGTA TCCAACCAGA 480
AAGTCACGGC TAACTACGTG CCAGCAGCCG CGGTAATACG TAGGTGGCAA GCGTTATCCG 540
GATTTATTGG GCGTAAAGCG AGCGCAGGCG GTTGCTTAGG TCTGATGTGA AAGCCTTTCG 600
GCTTAACCGA AGAAGTGCAT CGGAAACCGG GCGACTTGAG TGCAGAAGAG GACAGTGGAA 660
CTCCATGTGT AGCGGTGGAA TGCGTAGATA TATGGAAGAA CACCAGTGGC GAAGGCGGCT 720
GTCTGGTCTG CAACTGACGC TGAGGCTCGA AAGCATGGGT AGCGAACAGG ATTAGATACC 800
CTGGTAGTCC ATGCCGTAAA CGATGAGTGC TAGGTGTTGG AGGGTTTCCG CCCTTCAGTG 860
CCGGAGCTAA CGCATTAAGC ACTCCGCCTG GGGAGTACGA CCGCAAGGTT GAAACTCAAA 920
GGAATTGACG GGGGCCCGCA CAAGCGGTGG AGCATGTGGT TTAATTCGAA GCTACGCGAA 980
GAACCTTACC AGGTCTTGAC ATCTTGCGCT AACCTTAGAG ATAAGGCGTT CCCTTCGGGG 1040
ACGCAATGAC AGGTGGTGCA TGGTCGTCGT CAGCTCGTGT CGTGAGATGT TGGGTTAAGT 1100
CCCGCAACGA GCGCAACCCT TGTTACTAGT TGCCAGCATT AAGTTGGGCA CTCTAGTGAG 1160
ACTGCCGGTG ACAAACCGGA GGAAGGTGGG GACGACGTCA GATCATCATG CCCCTTATGA 1220
CCTGGGCTAC ACACGTGCTA CAATGGACGG TACAACGAGT CGCAAGCTCG CGAGAGTAAG 1280
CTAATCTCTT AAAGCCGTTC TCAGTTCGGA CTGTAGGCTG CAACTCGCCT ACACGAAGTC 1340
GGAATCGCTA GTAATCGCGG ATCAGCATGC CGCGGTGAAT ACGTTCCCGG GCCTTGTACA 1400
CACCGCCCGT CACACCATGG GAGTTTGTAA CGCCCAAAGT CGGTGGCCTA ACCTTTATGG 1460
AGGGAGCCGC CTAAGGCGGA CAGAT 1485
Claims (5)
1. The Lactobacillus reuteri strain is named as Lactobacillus reuteri HCS02-001, and is preserved in China general microbiological culture Collection center (CGMCC) at 27 days 04 in 2020 and with the preservation number of CGMCC No. 19746.
2. Use of a strain of lactobacillus reuteri according to claim 1 for the preparation of a cholesterol lowering medicament.
3. The use of a strain of lactobacillus reuteri according to claim 1 for the preparation of a medicament for inhibiting gardnerella vaginalis.
4. A pharmaceutical preparation with cholesterol-lowering and/or gardnerella vaginalis-inhibiting functions, which comprises an effective dose of the Lactobacillus reuteri strain of claim 1 and pharmaceutically acceptable excipients.
5. A bacterial agent having a function of lowering cholesterol and/or inhibiting Gardnerella vaginalis, which comprises the Lactobacillus reuteri according to claim 1.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116585362A (en) * | 2023-06-02 | 2023-08-15 | 江西仁仁健康微生态科技有限公司 | Application and product of lactobacillus reuteri HCS02-001 in uric acid reduction |
| CN116590357A (en) * | 2023-06-08 | 2023-08-15 | 江西仁仁健康微生态科技有限公司 | Application of lactobacillus reuteri in production of gamma-aminobutyric acid and sleep-aiding products |
| CN116622570A (en) * | 2023-05-22 | 2023-08-22 | 微康益生菌(苏州)股份有限公司 | Lactobacillus reuteri LR08, and application, product and method thereof in preparation of medicines for preventing and treating vaginitis |
| CN116649578A (en) * | 2022-12-16 | 2023-08-29 | 江西仁仁健康微生态科技有限公司 | Metagen, preparation method thereof and application thereof in antibacterial products |
| CN116751705A (en) * | 2023-05-17 | 2023-09-15 | 上海华朴生命健康科技有限公司 | Lactobacillus reuteri with effect of improving colpitis and application thereof |
| CN117286083A (en) * | 2023-11-24 | 2023-12-26 | 杭州微致生物科技有限公司 | Lactobacillus reuteri VB319 and culture device and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105219683A (en) * | 2015-11-04 | 2016-01-06 | 广东省农业科学院动物科学研究所 | One strain has L. reuteri strain and the application thereof of prebiotic characteristics |
| CN107267415A (en) * | 2017-06-23 | 2017-10-20 | 广东龙创基药业有限公司 | A kind of lactobacillus reuteri and its application for preparing vagina antibacterial medicines |
| CN112358999A (en) * | 2020-11-26 | 2021-02-12 | 中国农业大学 | Lactobacillus reuteri and application thereof |
| JP2021065223A (en) * | 2019-10-28 | 2021-04-30 | 葡萄王生技股▲ふん▼有限公司 | Use of lactobacillus reuteri strain gkr1 for preparation of composition to reduce urine acid |
| WO2021250284A1 (en) * | 2020-06-12 | 2021-12-16 | Eligo Bioscience | Specific decolonization of antibiotic resistant bacteria for prophylactic purposes |
-
2022
- 2022-05-25 CN CN202210572722.9A patent/CN114990011B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105219683A (en) * | 2015-11-04 | 2016-01-06 | 广东省农业科学院动物科学研究所 | One strain has L. reuteri strain and the application thereof of prebiotic characteristics |
| CN107267415A (en) * | 2017-06-23 | 2017-10-20 | 广东龙创基药业有限公司 | A kind of lactobacillus reuteri and its application for preparing vagina antibacterial medicines |
| JP2021065223A (en) * | 2019-10-28 | 2021-04-30 | 葡萄王生技股▲ふん▼有限公司 | Use of lactobacillus reuteri strain gkr1 for preparation of composition to reduce urine acid |
| WO2021250284A1 (en) * | 2020-06-12 | 2021-12-16 | Eligo Bioscience | Specific decolonization of antibiotic resistant bacteria for prophylactic purposes |
| CN112358999A (en) * | 2020-11-26 | 2021-02-12 | 中国农业大学 | Lactobacillus reuteri and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| SIWEN YANG ET AL.: "Effect of Oral Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on the Vaginal Microbiota, Cytokines and Chemokines in Pregnant Women", 《NUTRIENTS》 * |
| 余萍等: "具有降胆固醇功能的罗伊氏乳杆菌的筛选及其益生作用", 《现代食品科技》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116649578A (en) * | 2022-12-16 | 2023-08-29 | 江西仁仁健康微生态科技有限公司 | Metagen, preparation method thereof and application thereof in antibacterial products |
| CN116751705A (en) * | 2023-05-17 | 2023-09-15 | 上海华朴生命健康科技有限公司 | Lactobacillus reuteri with effect of improving colpitis and application thereof |
| CN116751705B (en) * | 2023-05-17 | 2024-03-15 | 上海华朴生命健康科技有限公司 | Lactobacillus reuteri with effect of improving colpitis and application thereof |
| CN116622570A (en) * | 2023-05-22 | 2023-08-22 | 微康益生菌(苏州)股份有限公司 | Lactobacillus reuteri LR08, and application, product and method thereof in preparation of medicines for preventing and treating vaginitis |
| CN116622570B (en) * | 2023-05-22 | 2023-11-03 | 微康益生菌(苏州)股份有限公司 | Lactobacillus reuteri LR08, and application, product and method thereof in preparation of medicines for preventing and treating vaginitis |
| CN116585362A (en) * | 2023-06-02 | 2023-08-15 | 江西仁仁健康微生态科技有限公司 | Application and product of lactobacillus reuteri HCS02-001 in uric acid reduction |
| CN116590357A (en) * | 2023-06-08 | 2023-08-15 | 江西仁仁健康微生态科技有限公司 | Application of lactobacillus reuteri in production of gamma-aminobutyric acid and sleep-aiding products |
| CN117286083A (en) * | 2023-11-24 | 2023-12-26 | 杭州微致生物科技有限公司 | Lactobacillus reuteri VB319 and culture device and application thereof |
| CN117286083B (en) * | 2023-11-24 | 2024-03-22 | 杭州微致生物科技有限公司 | Lactobacillus reuteri VB319 and culture device and application thereof |
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