CN105087436A - High-producing 3-hydroxypropionaldehyde strain and application thereof to prevention and treatment of vaginal infection of mammals - Google Patents
High-producing 3-hydroxypropionaldehyde strain and application thereof to prevention and treatment of vaginal infection of mammals Download PDFInfo
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Abstract
The invention discloses a high-producing 3-hydroxypropionaldehyde strain and application thereof to prevention and treatment of vaginal infection of mammals, and relates to a microorganism and a composition thereof, in particular to a microorganism and a composition thereof for prevention and treatment of vaginal infection of the mammals. Further, the invention aims to provide lactobacillus reuteri short in growth cycle and high in probiotics and application of the lactobacillus reuteri to prevention and treatment of vaginal infection of the mammals. The high-producing 3-hydroxypropionaldehyde strain refers to the lactobacillus reuteri RS01 and is preserved with the preservation number CGMCC No.9971 in China General Microbiological Culture Collection Center on November 14th, 2014. The high-producing 3-hydroxypropionaldehyde strain has the advantages that the high-producing 3-hydroxypropionaldehyde strain is cultured easily and high in target product yield, and fermentation liquor of the high-producing 3-hydroxypropionaldehyde strain is safe and free of toxic and side effects and has broad-spectrum antibacterial effect. The high-producing 3-hydroxypropionaldehyde strain is applied to the field of prevention and treatment of vaginal infection of the mammals.
Description
Technical field
The present invention relates to a kind of microorganism and composition thereof, particularly relating to a kind of microorganism for preventing and treating manmmal vagina infection and composition thereof.
Background technology
Prebiotic rhzomorph 3-HPA (3-HPA), CAS:2134-29-4, molecular formula C
3h
6o
2, dissolve in ethanol, ether, acetone, very easily water-soluble, be easily polymerized, belong to aldol compounds, Nature comparison is active, free from extraneous odour.Prebiotic rhzomorph 3-HPA mainly presents 3-HPA monomer, hydrate and Cyclodimerization body three kinds of states in aqueous.3-HPA has aldehyde radical and hydroxyl, the hydroxyl reaction relevant with aldehyde radical easily occurs, easy condensation.
Prebiotic rhzomorph 3-HPA can be produced by the bacterial metabolism glycerine surely grown in people and animal intestinal, has potential use widely in its precursor at biological sterilization agent, anti-infective therapy's agent, Biological cross-linker, novel biomaterial etc.
The main component of Luo Yishi element is the monomer of 3-HPA, hydrate and Cyclodimerization body.Research shows, the Luo Yishi element of 15-30 μ g/ml just can suppress the growth of gram-positive microorganism, Gram-negative bacteria, yeast, fungi and protozoon etc.Because Luo Yishi element had both had a broad-spectrum antimicrobial characteristic, harmless to humans and animals again, its purposes had just progressively been widely deployed, as biological preservative, for extending the commodity price phase of sausage, frozen product etc.; As biological sterilization agent, for can not the Milk of high-temperature sterilization, the sterilizing of biomaterial; As chewing gum additive, for killing Oral Pathogens preventing dental caries; As anti-infective therapy's agent, substitute antibiotics is used for the diseases induced treatment of zoonosis protozoon.
Prebiotic rhzomorph 3-HPA monomer is the alcohol aldehyde compound of three carbon, can be formed crosslinked with the amido reaction in protein.In acid condition, three prebiotic rhzomorph 3-HPA monomers form acetal, can carry out dialdehyde base crosslinking reaction with free amino; And in the basic conditions, single cross-link reaction of formaldehyde may be similar to.Therefore can as novel crosslinker, the biological tissue being cross-linked the denaturation temperature of artifact tissue, crosslinking index, resistance to enzymolysis ability and mechanical property etc. crosslinked with glutaraldehyde (glutaraldehyde) is suitable, but its cytotoxicity is lower.Therefore can the glutaraldehyde of substituted chemistry synthesis and epoxy compounds as novel Biological cross-linker, thus overcome the too high and immune response of the hardening of tissue that chemical cross-linking agent process biological tissue runs into, calcification, cytotoxicity and cross the problems such as strong.
In addition, prebiotic rhzomorph 3-HPA also can be used as multiple emerging chemical as the precursor of propenal, vinylformic acid, 1,3-PD etc., for the preparation of novel polymer material.
It is common disease and the frequently-occurring disease of women that gynaecology's V&V catches, and its sickness rate rises year by year, and chlorhexidine acetate and metronidazole compound preparation are the common drugs for the treatment of this kind of disease.These preparations have Chlorhexidine Compound, metronidazole,clotrimazole and chlorhexidine acetate suppositories agent etc., and wherein chlorhexidine acetate is 8mg/ grain, and the content of metronidazole is 200mg/ grain, are used for the treatment of the various vaginitis that the pathogenic agent such as various bacterium, anerobe, mould, trichomonad cause.But chlorhexidine acetate/metronidazole compound preparation is the successful when treating early stage catching just, and be not very desirable in the heavier effect that catches for the treatment of, and chlorhexidine acetate/metronidazole compound preparation is treating colpitic while, demonstrate larger untoward reaction, show as the local excitation of vagina comparatively large, produce the red and swollen phenomenon in local; And the use of these medicines also can suppress intravaginal normal microflora active, thus destroys the normal eubiosis of intravaginal.
Summary of the invention
The technical problem to be solved in the present invention is to provide the high lactobacillus reuteri of short, the prebiotic rhzomorph output of a kind of growth cycle, and the application in control manmmal vagina infection.
The present invention one strain 3-HPA (prebiotic rhzomorph) superior strain, it is characterized in that: it is lactobacillus reuteri (Lactobacillusreuteri) RS01, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, preserving number is CGMCCNo.9971, and preservation date is on November 14th, 2014.
Further, 3-HPA superior strain of the present invention, the cell walls dyeing of described lactobacillus reuteri RS01 is in Gram-positive, and without gemma, thalline presents corynebacterium, is distributed as single or in connection shape.
Further, 3-HPA superior strain of the present invention, the long 1.5-3um of described lactobacillus reuteri RS01 thalline, wide 0.6-0.9um.
Further, 3-HPA superior strain of the present invention, the smooth exquisiteness of bacterium colony of described lactobacillus reuteri RS01, neat in edge, bacterium colony size uniformity is oyster white, moistening, opaque.
A kind of method utilizing lactobacillus reuteri RS01 to produce 3-HPA of the present invention, comprises the following steps:
One, prepare substratum: peptone 10g, extractum carnis 10g, glucose 20g, yeast extract 5g, tween 80 1ml, dipotassium hydrogen phosphate 2g, Triammonium citrate 2g, sodium acetate 5g, magnesium sulfate 0.5g, manganous sulfate 0.2g, Cys 0.5g, sodium thioglycollate 0.05g, VB120.005g, glycerine 3.68g add water and be settled to 1L, adjust ph is 6.8;
Two, above-mentioned lactobacillus reuteri RS01 is accessed in the substratum of preparation in step one, 37 DEG C, dissolved oxygen amount is under the condition of 0, anaerobically fermenting 5h;
Three, be separated: first time is separated and adopts sheet frame to be separated, collect thalline and cause thalline dead fast and not easily; Then the thalline of separation is accessed in new substratum, 37 DEG C, under dissolved oxygen amount is the condition of 0, cultivate 4h; Second time is separated and adopts high speed tubular-bowl centrifuge, and object product can be made effectively to be separated with thalline;
Four, the fermented liquid that twice separation obtains is merged.
In the present invention, above-mentioned bacterial classification be have employed to the method for same bacterial classification fermenting twice, obtain the 3-HPA of high density.
The application of a kind of 3-HPA superior strain of the present invention in control manmmal vagina infection, is characterized in that: for the preparation of the medicine of various treatment vaginitis or prevention of urogenital infections.
The present invention one probiotics element superior strain and the application difference from prior art in control manmmal vagina infection thereof are:
1, be easy to cultivate: present invention employs anaerobically fermenting, control the oxygen concentration in fermenting process; Namely 3-HPA is transformed, to the 3-HPA of high density by fermenting twice.
2, output is high: the lactobacillus reuteri RS01 in the present invention is after mutagenesis screening, and growth cycle time half, target product (3-HPA) output is promoted to 2.5-2.8g/L by being originally almost 0.
3, energy-conservation: the lactobacillus reuteri RS01 in the present invention, after mutagenesis screening, because the shortening of growth cycle is over half, thus greatly reduces production cost, has saved water, electricity, vapour equal energy source.
4, safely, have no side effect: adopt the fermented liquid of lactobacillus reuteri RS01 of the present invention to carry out gavage and vaginal injection to SD system rat, all checking has no side effect.
5, has a broad antifungal spectrum: prebiotic rhzomorph superior strain meta-bolites of the present invention all has good result for the treatment of to the vaginitis that bacterium, fungi, protozoon cause, and proves that it has the anti-microbial effect of wide spectrum, can be used for prevention or treats mammiferous vaginal infection.
Accompanying drawing explanation
Fig. 1 is the microscopy figure of lactobacillus reuteri RS01 of the present invention;
Fig. 2 is the bacterium colony figure of lactobacillus reuteri RS01 of the present invention;
Fig. 3 be lactobacillus reuteri RS01 of the present invention press down Candida albicans figure;
Fig. 4 is that the streptococcus aureus that presses down of lactobacillus reuteri RS01 of the present invention is schemed;
Fig. 5 is that the intestinal bacteria that press down of lactobacillus reuteri RS01 of the present invention are schemed.
Embodiment
With the following Examples with proof test to a probiotics element superior strain of the present invention and be further described preventing and treating the application in manmmal vagina infection.
Lactobacillus reuteri RS01 of the present invention is the forward mutation bacterial strain that space flight filters out.Under the condition of High energy particles Radiation, microgravity, alternating magnetic field, there is the multiple bacterial strain with the generation of metabolism in various degree 3-HPA ability in lactobacillus reuteri, by carrying out the bactericidal assay of different gradient to pathogenic flora, screening obtains the bacterial strain of sterilizing rate more than 90% in 60min, prebiotic rhzomorph (3-HPA) superior strain namely in the present invention.
Fig. 1 is the microscopy figure of lactobacillus reuteri RS01 of the present invention; Fig. 2 is the bacterium colony figure of lactobacillus reuteri RS01 of the present invention.
Embodiment 1
The method of the production 3-HPA of prebiotic rhzomorph superior strain in the present invention, carry out according to the following steps:
One, prepare substratum: peptone 10g, extractum carnis 10g, glucose 20g, yeast extract 5g, tween 80 1ml, dipotassium hydrogen phosphate 2g, Triammonium citrate 2g, sodium acetate 5g, magnesium sulfate 0.5g, manganous sulfate 0.2g, Cys 0.5g, sodium thioglycollate 0.05g, VB120.005g, glycerine 3.68g add water and be settled to 1L, adjust ph is 6.8;
Two, lactobacillus reuteri RS01 of the present invention is accessed in the substratum of preparation in step one, 37 DEG C of anaerobically fermenting 5h;
Three, be separated: first time is separated and adopts sheet frame to be separated, collect thalline and cause thalline dead fast and not easily; Then the thalline of separation is accessed in new substratum, 37 DEG C, under dissolved oxygen amount is the condition of 0, cultivate 4h; Second time is separated and adopts high speed tubular-bowl centrifuge, and object product can be made effectively to be separated with thalline;
Four, the fermented liquid that twice separation obtains is merged.
According to 3-HPA thermal dehydration formation propenal under strongly acidic conditions, and tryptophane is under the existence of concentrated hydrochloric acid, generates purple material with acrolein reaction, under 560nm, have absorption peak.Therefore, the output of gained 3-HPA is detected by color reaction.After measured, the amount that 3-HPA is produced in the lactobacillus reuteri fermentation before sudden change is almost 0, and the lactobacillus reuteri RS01 in employing the application is in conjunction with above-mentioned fermentation process, and the output of gained 3-HPA is 2.5-2.8g/L; And the growth cycle of bacterial classification foreshortens to present 5h by original 12-20h, greatly reduce production cost, save water, electricity, vapour equal energy source.
Bacteriostatic test
Be diluted to tetraploid amass being separated the fermented liquid obtained in embodiment 1, according to following steps, the pressing down of fermented liquid/sterilization (Candida albicans, streptococcus aureus, intestinal bacteria) ability is measured:
One, substratum is prepared: preparation nutrient agar;
Two, sterilizing: substratum step one prepared, culture dish, Oxford cup, liquid-transfering gun rifle is first-class puts into Autoclave sterilizing;
Three, getting the pathogenic bacterium access activated in right amount is cooled in the substratum of 40 DEG C, obtained bacteria suspension;
Four, the bacteria suspension in step 3 is poured in culture dish, treat culture medium solidifying;
Five, in culture dish, put into Oxford cup (each plate puts 3), then add 0.24ml testing sample in the cup of Oxford;
Six, the culture dish in step 5 is put into incubator and cultivate 12h;
Seven, inhibition zone size is observed.
Fungistatic effect judges according to inhibition zone size, and standard is as shown in table 1, and acquired results is shown in Fig. 3-5.
The criterion of table 1 fungistatic effect
Bacteriostatic test result shows, the antibacterial circle diameter that fermented liquid of the present invention suppresses Candida albicans to be formed is 26.3mm, the antibacterial circle diameter suppressing streptococcus aureus to be formed is 27.8mm, the antibacterial circle diameter suppressing intestinal bacteria to be formed is 31.2mm, result is all significantly higher than extremely sensitive standard, above method validation is adopted to dilute the fungistatic effect of the fermented liquid of 80 times of volumes, find that fermented liquid is diluted to 80 times and still has fungistatic effect, illustrate that the fermented liquid of prebiotic rhzomorph superior strain of the present invention is to Candida albicans, streptococcus aureus and intestinal bacteria all have very strong killing action.
Safety testing
For the security of checking the present invention prebiotic rhzomorph superior strain, adopt fermented liquid of the present invention to carry out gavage and vaginal injection to SD system rat, study prebiotic rhzomorph superior strain metabolism of the present invention and produce the impact of thing on its blood biochemistry index and vagina tissue.
Get the SD female system rat 30 that body weight is 200g ± 20g, be divided into three groups, negative control group 10, blank group 10, experimental group 10.The extracting solution gavage that experimental group obtains by 1mL embodiment 1, the extracting solution obtained by 0.2mL embodiment 1 injects vagina, 1 time/d, altogether 21d.
Acquired results is as follows:
1, the serum alanine aminotransferase (ALT) of experimental group, serum creatinine (CREA), serum uric acid (uA), total serum protein (TP), serum albumin (ALB), serum globulin (GLOB) are not all higher than the index of Normal group.
2, tissue examination result is, liver kidney, vagina tissue paraffin section, and HE dyes, microscopic examination, has no distortion infiltration, red and swollen, inflammation.
3, vaginal irritation test-results is, has no cytomorphosis and leukocyte infiltration, redness, hyperemia, hemorrhage, inflammation, festers, without obvious pathological change.
Effect experiment
In order to verify that the present invention's prebiotic rhzomorph superior strain meta-bolites is to the result for the treatment of of bacterial vaginitis, colpitis mycotica and trichomonal vaginitis, adopt extracting solution obtained in embodiment 1 to the administration of SD rat vagina.
Use Candida albicans bacteria suspension modeling Candida albicans vaginitis SD rat, and 6d gets vaginal swab smear poststaining microscopy after modeling, the classification of vagina cleanliness, carries out bacteriological detection, determines that modeling is set up.32 of modeling female sd inbred rats are divided into 4 groups at random, often organize 8, add blank group and have 5 groups altogether, contrived experiment grouping is as shown in table 2.Administration group gives corresponding trial-product respectively by the dosage of table 2, and blank group and model control group give the sample of the embodiment 1 after diluting with equal-volume, successive administration 7d once a day.Observe growth of animal, activity and the clean situation of vulva, check vagina cleanness degree, carry out bacteriological detection and get vagina tissue carrying out pathological examination.
The experiment grouping of drug effect verified by table 2
Judging criterion: vagina cleanness degree classification and vaginitis take a turn for the better, criterion of cure, with reference to " obstetrics and gynecology " the 5th edition state-compiled textbook; Modeling success judging criterion, with reference to " pharmacological experimental methodology "; Curative ratio and efficient calculation formula as follows:
Number of animals/total number of animals × 100 of each group of curative ratio (%)=recovery from illness
Number of animals/total number of animals × 100 of each group of efficient (%)=improvement
Experimental result
Visual results: after modeling, obvious inflammatory reaction all appears in animal: the hyperemia of vagina periphery, oedema, white secretory product increase; Blank treated animal pudendum cleaning, secretory product without exception.Through medication after 7 days, trial-product treated animal vaginal congest, oedema etc. all have alleviation in various degree, but the reaction of model group animal local inflammation is still more obvious.
Vagina cleanness degree change and efficacy determination after treatment: result is as shown in table 3.
Table 3 embodiment 1 obtains product to SD rat colpitis mycotica result for the treatment of
As shown in Table 3, before treatment outside blank group, all the other are respectively organized vagina cleanness degree and are III or IV, and after using the embodiment of the present invention 1 to obtain product, compared with model control group, vagina cleanness degree and curative ratio are all significantly improved.
Histopathology: model control group SD rat vagina epithelium thickens, and level increases, the visible strip epithelial cell necrosis of epithelial surface and epithelial cell shedding, epithelium is supervised and visible more cell infiltration or stove shape cell infiltration in upper subcutaneous tunica propria.After using the embodiment of the present invention 1 product, the minority epithelial cell sex change of low dose group rarely seen vagina epithelium surface, downright bad individually, tunica propria has no inflammatory reaction.Middle dosage group has a little cell infiltration stove under only having a SD rat vagina epithelium mucous membrane, and epithelial structure is complete, has no epithelial cell sex change, necrosis.High dose group vaginal epithelial cell structural integrity, is methodically arranged, there are no obvious inflammatory cell infiltration.
Use streptococcus aureus suspension modeling streptococcus aureus vaginitis SD rat, and 6d gets vaginal swab smear poststaining microscopy after modeling, the classification of vagina cleanliness, carries out bacteriological detection, determines that modeling is set up.32 of modeling female sd inbred rats are divided into 4 groups at random, often organize 8, add blank group and have 5 groups altogether, contrived experiment grouping is as shown in table 4.Administration group gives corresponding trial-product respectively by the dosage of table 4, and blank group and model control group give the sample of the embodiment 1 after diluting with equal-volume, successive administration 7d once a day.Observe growth of animal, activity and the clean situation of vulva, check vagina cleanness degree, carry out bacteriological detection and get vagina tissue carrying out pathological examination.
The experiment grouping of drug effect verified by table 4
Judging criterion is all identical with standard described in the above-mentioned checking Candida albicans vaginitis test of pesticide effectiveness.
Experimental result
Visual results: after modeling, obvious inflammatory reaction all appears in animal: the hyperemia of vagina periphery, oedema, secretory product is yellow, in water sample; Blank treated animal pudendum cleaning, secretory product without exception.Through medication after 7 days, trial-product treated animal vaginal congest, oedema etc. all have alleviation in various degree, but the reaction of model group animal local inflammation is still more obvious.
Vagina cleanness degree change and efficacy determination after treatment: result is as shown in table 5.
Table 5 embodiment 1 obtains product to SD rat colpitis mycotica result for the treatment of
As shown in Table 5, before treatment outside blank group, all the other are respectively organized vagina cleanness degree and are III or IV, and after using the embodiment of the present invention 1 to obtain product, compared with model control group, vagina cleanness degree and curative ratio are all significantly improved.
Histopathology: model control group SD rat vagina epithelium thickens, and level increases, the visible strip epithelial cell necrosis of epithelial surface and epithelial cell shedding, epithelium is supervised and visible more cell infiltration or stove shape cell infiltration in upper subcutaneous tunica propria.After using the embodiment of the present invention 1 product, the minority epithelial cell sex change of low dose group rarely seen vagina epithelium surface, downright bad individually, tunica propria has no inflammatory reaction.Middle dosage group has a little cell infiltration stove under only having a SD rat vagina epithelium mucous membrane, and epithelial structure is complete, has no epithelial cell sex change, necrosis.High dose group vaginal epithelial cell structural integrity, is methodically arranged, there are no obvious inflammatory cell infiltration.
Use Trichomonas vaginalis suspension modeling trichomonal vaginitis SD rat, and 6d gets vaginal swab smear poststaining microscopy after modeling, the classification of vagina cleanliness, carries out bacteriological detection, determines that modeling is set up.32 of modeling female sd inbred rats are divided into 4 groups at random, often organize 8, add blank group and have 5 groups altogether, contrived experiment grouping is as shown in table 6.Administration group gives corresponding trial-product respectively by the dosage of table 6, and blank group and model control group give the sample of the embodiment 1 after diluting with equal-volume, successive administration 7d once a day.Observe growth of animal, activity and the clean situation of vulva, check vagina cleanness degree, carry out bacteriological detection and get vagina tissue carrying out pathological examination.
The experiment grouping of drug effect verified by table 6
Judging criterion is all identical with standard described in the above-mentioned checking Candida albicans vaginitis test of pesticide effectiveness.
Experimental result
Visual results: after modeling, obvious inflammatory reaction all appears in animal: the hyperemia of vagina periphery, oedema, secretory product yellow-green colour, leukorrhea is thin and have stench; Blank treated animal pudendum cleaning, secretory product without exception.Through medication after 7 days, trial-product treated animal vaginal congest, oedema etc. all have alleviation in various degree, but the reaction of model group animal local inflammation is still more obvious.
Vagina cleanness degree change and efficacy determination after treatment: result is as shown in table 7.
Table 7 embodiment 1 obtains product to SD rat colpitis mycotica result for the treatment of
As shown in Table 7, before treatment outside blank group, all the other are respectively organized vagina cleanness degree and are III or IV, and after using the embodiment of the present invention 1 to obtain product, compared with model control group, vagina cleanness degree and curative ratio are all significantly improved.
Histopathology: model control group SD rat vagina epithelium thickens, and level increases, the visible strip epithelial cell necrosis of epithelial surface and epithelial cell shedding, epithelium is supervised and visible more cell infiltration or stove shape cell infiltration in upper subcutaneous tunica propria.After using the embodiment of the present invention 1 product, the minority epithelial cell sex change of low dose group rarely seen vagina epithelium surface, downright bad individually, tunica propria has no inflammatory reaction.Middle dosage group has a little cell infiltration stove under only having a SD rat vagina epithelium mucous membrane, and epithelial structure is complete, has no epithelial cell sex change, necrosis.High dose group vaginal epithelial cell structural integrity, is methodically arranged, there are no obvious inflammatory cell infiltration.
Result shows to adopt prebiotic rhzomorph superior strain meta-bolites of the present invention to treat bacterial vaginitis, colpitis mycotica and trichomonal vaginitis, compared with control group, the equal result for the treatment of of prebiotic rhzomorph superior strain meta-bolites of the present invention high, medium and low dosage group is remarkable.Illustrate that prebiotic rhzomorph superior strain meta-bolites of the present invention all has good result for the treatment of to the vaginitis that bacterium, fungi, protozoon cause, prove that it has the anti-microbial effect of wide spectrum, therefore can be used for prevention or treat mammiferous vaginal infection.
Above-described embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various distortion that those of ordinary skill in the art make technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determines.
Claims (6)
1. a strain 3-HPA superior strain, is characterized in that: it is lactobacillus reuteri RS01, and be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCCNo.9971, and preservation date is on November 14th, 2014.
2. 3-HPA superior strain according to claim 1, is characterized in that: the dyeing of the cell walls of described lactobacillus reuteri RS01 is in Gram-positive, and without gemma, thalline presents corynebacterium, is distributed as single or in connection shape.
3. 3-HPA superior strain according to claim 1, is characterized in that: the long 1.5-3um of described lactobacillus reuteri RS01 thalline, wide 0.6-0.9um.
4. 3-HPA superior strain according to claim 1, is characterized in that: the smooth exquisiteness of bacterium colony of described lactobacillus reuteri RS01, neat in edge, bacterium colony size uniformity, oyster white, moistening, opaque.
5. utilize the RS01 of lactobacillus reuteri described in claim 1 to produce a method for 3-HPA, it is characterized in that comprising the following steps:
One, prepare substratum: peptone 10g, extractum carnis 10g, glucose 20g, yeast extract 5g, tween 80 1ml, dipotassium hydrogen phosphate 2g, Triammonium citrate 2g, sodium acetate 5g, magnesium sulfate 0.5g, manganous sulfate 0.2g, Cys 0.5g, sodium thioglycollate 0.05g, VB120.005g, glycerine 3.68g add water and be settled to 1L, adjust ph is 6.8;
Two, lactobacillus reuteri RS01 described in claim 1 is accessed in the substratum of preparation in step one, 37 DEG C, dissolved oxygen amount is under the condition of 0, cultivates 5h;
Three, be separated: first time is separated and adopts sheet frame to be separated; Then the thalline of separation is accessed in new substratum, 37 DEG C, dissolved oxygen amount is under the condition of 0, cultivates 4h; Second time is separated and adopts high speed tubular-bowl centrifuge;
Four, the fermented liquid that twice separation obtains is merged.
6. the application of 3-HPA superior strain in control manmmal vagina infection, is characterized in that: for the preparation of the medicine of various treatment vaginitis or prevention of urogenital infections.
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| 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 |
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| CN103114051A (en) * | 2012-11-15 | 2013-05-22 | 上海交大昂立股份有限公司 | Roy bacterium lacticum and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
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Application publication date: 20151125 |