CN103710290A - Probiotics lactobacillus rhamnosus hsryfm1301 originated in Guangxi Bama longevity village and application of probiotics lactobacillus rhamnosus hsryfm1301 - Google Patents
Probiotics lactobacillus rhamnosus hsryfm1301 originated in Guangxi Bama longevity village and application of probiotics lactobacillus rhamnosus hsryfm1301 Download PDFInfo
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Abstract
The invention relates to a probiotics lactobacillus rhamnosus hsryfm1301 originated in a Guangxi Bama longevity village and an application of the probiotics lactobacillus rhamnosus hsryfm1301. The bacterium has the ability of decomposing cholesterol and triglyceride, and the preservation number of the bacterium is CGMCC (China General Microbiological Culture Collection Center) NO.8545. The separation, the screening and the identification of the lactobacillus rhamnosus hsryfm1301 comprise the following steps: separating lactic acid bacteria from intestinal tracts of Guangxi Bama longevity people and screening out bacterial strains with relatively strong ability of decomposing the cholesterol and the triglyceride through in vitro tests; on the basis of comprehensively comparing the acid resisting characteristic, the cholate resisting characteristic, the antibacterial characteristic and the antibiotic sensibility, obtaining a probiotics strain hsryfm1301 with the most excellent probiotic performances; and through physiological and biochemical tests, API (Analytic Products INC) reagent strip and 16s rDNA (ribosomal Deoxyribose Nucleic Acid) sequencing analyses, identifying the screened strain as the lactobacillus rhamnosus.
Description
Technical field
The present invention relates to biological technical field, particularly a kind of probiotic bacterium lactobacillus rhamnosus hsryfm1301 and application thereof that comes from Bama of Guangxi Changshou village.
Technical background
Serum cholesterol level is too high is the major cause of cardiovascular disorder, and medicine is the major way of current Cardiovarscular, but the cost of pharmacological agent is higher and have a larger side effect.Clinical study is found, reduces the mortality ratio that cholesterol in serum level can significantly reduce cardiovascular diseases.A large amount of animal and human's bodies are tested and are shown, milk-acid bacteria can reduce the cholesterol levels in serum.The actives mass-energy that milk-acid bacteria metabolism produces suppresses the growth of pathogenic bacterium in enteron aisle, the microecological balance of regulating intestinal canal; Bacterium in gi tract, by the horizontal transfer of antibiotics resistance gene, has brought a serious difficult problem to clinical medicine; Therefore cholesterol and triglyceride level are decomposed in screening, and have bacteriostasis and the lower probiotic bacterium of resistance has great significance.
Summary of the invention
The object of the invention is to provide has probiotic strain and the application thereof of decomposing cholesterol and triglyceride level ability.
The invention provides a strain lactobacillus rhamnosus (Lactobacillus rhamnosus) hsryfm1301, this bacterium has the cholesterol of decomposition and triglyceride level ability, on December 6th, 2013, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preserving number is CGMCC NO.8545.
Through evidence, this bacterial strain has the ability of decomposing cholesterol and triglyceride level, and resistance to simulated gastric fluid and cholate, have stronger bacteriostasis, and resistance is lower simultaneously; By Physiology and biochemistry, API reagent strip and 16S rDNA sequence alignment, analyzing, is lactobacillus rhamnosus by this identification of strains.
The present invention also provides lactobacillus rhamnosus hsryfm1301 to have the application in fermented-milk, leben and the active bacteria formulation that decomposes cholesterol and triglyceride level ability in preparation.
Lactobacillus rhamnosus hsryfm1301 of the present invention has the ability of decomposing cholesterol and triglyceride level, and resistance to simulated gastric fluid and cholate, there is stronger bacteriostasis and resistance lower simultaneously, use it for and prepare fermented-milk, leben and active bacteria formulation, there is good prospect.
Accompanying drawing explanation
Fig. 1 is the aspect graph of bacterial strain of the present invention.
Fig. 2 is strains A PI reagent strip evaluation figure of the present invention.
Fig. 3 is strains A PI qualification result of the present invention.
Fig. 4 is bacterial strain 16S rDNA pcr amplification product electrophorogram of the present invention; Swimming lane M:100+2+3Kb DNA maker; Swimming lane 1 is: hsryfm130116S rDNA amplified production.
Embodiment
This research adopts traditional spread plate partition method separating lactic acid bacterium from the sample of Bama of Guangxi Changshou village collection, by in vitro tests, filter out and decompose cholesterol and the stronger lactic bacterium strains of triglyceride level ability, then by simulation human gastrointestinal tract environmental screening, go out the stronger bacterial strain of acidproof bile tolerance ability, and study its biocidal property and resistance, by comprehensive comparison, obtain the probiotic strain that prebiotic performance is the most excellent.
Relevant substratum and agent prescription
(1) MRS-THIO substratum: peptone 10.0g, Sodium acetate trihydrate 3.0g, extractum carnis 8.0g, Triammonium citrate 2.0g, yeast extract 4.0g, magnesium sulfate heptahydrate 0.2g, glucose 10.0g, manganese sulfate monohydrate 0.04g, tween-80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium thioglycollate 2.0g, adjust pH to 6.4,121 ℃ of sterilizing 15min, cooling standby.
(3) cholesterol substratum: cholesterol 0.10g, tween-80 1.0mL, sucrose ester 0.10g, glacial acetic acid 5.0mL, ice-bath ultrasonic fragmentation is standby.The cholesterol micellar solution of above-mentioned preparation is joined in MRS liquid nutrient medium, and the final concentration that makes cholesterol in substratum is 0.10mg/mL.And adjust pH to 6.50 ± 0.20, and 121 ℃ of sterilizing 15min, cooling standby.
(4) triglyceride level substratum: 2% polyvinyl alcohol water solution is mixed in the ratio of 3:1 with vegetables oil, after processing with high-speed tissue mashing machine as the source of triglyceride level.The vegetables oil emulsification liquid of above-mentioned preparation is joined in MRS liquid nutrient medium, adjust pH to 6.50 ± 0.20,121 ℃ of sterilizing 15min, cooling standby.
(5) preparation of simulated gastric fluid: the HCl that accurately measures 20.0mL1mol/L, with deionized water, pH is adjusted to respectively to 2.0,3.0, add NaCl to make its massfraction reach 0.2%, in every 100.0mL solution, add 1.0g stomach en-, after fully dissolving, with 0.22 μ m millipore filtration filtration sterilization in aseptic operating platform, in aseptic reagent bottle, cryopreservation is standby.
(6) ferric ammonium sulfate developer: take 4.46g ferric ammonium sulfate and be dissolved in 100.0mL85% phosphoric acid, get 10.0mL solution and be settled to 100.0mL with the vitriol oil, put into silica dehydrator ware standby.
Concrete steps:
1 sample collecting
Gather long-lived crowd and the crowd of family sample thereof in Bama of Guangxi Changshou village, add after the sealing of 1mL sterilising liq paraffin oil, be placed in rapidly in ice chest the milk-acid bacteria in sample separation.
The separation of 2 milk-acid bacterias
The sample gathering, after gradient dilution, is inoculated in respectively in MRS solid medium, LBS solid medium, and 37 ℃ of anaerobism are cultivated 48h, the colonies typical on picking flat board, and line separation obtains pure bacterium colony.Pure bacterium colony on each flat board of picking is in MRS liquid nutrient medium, and 37 ℃ of anaerobism are cultivated after 24h, the standby or freeze-drying preservation of 4 ℃ of refrigerator cold-storages.
The physiological and biochemical test of 3 milk-acid bacterias
The milk-acid bacteria being separated to is carried out to gramstaining, in micro-Microscopic observation thalli morphology, and carry out the physiological and biochemical tests such as catalase, mobility, nitrate reduction and indoles.
Result shows, from 15 duplicate samples that gather, the separated 217 strain bacterial strains that obtain, find that by physiological and biochemical test 156 strain bacterium gramstaining are positive, are shaped as shaft-like, rod-short or spherical, under 15 ℃ and 45 ℃ of environment, all can grow, catalase, mobility, nitrate reduction, indoles, product H
2s and gelatin liquification test are all negative, so 156 strain separation of bacterial are initially identified as to milk-acid bacteria.
4 milk-acid bacterias are decomposed the mensuration of cholesterol and triglyceride level ability
The milk-acid bacteria having activated is seeded in MRS-THIO cholesterol substratum by 3% inoculum size, cultivate after 24h for 37 ℃, get nutrient solution 0.2mL, add the vibration of 1.0mL dehydrated alcohol to mix 1min, add the vibration of 3.8mL dehydrated alcohol to mix 1min, after standing 5min, vibration mixes again, the centrifugal 10min of 4000r/min again, get 2.0mL supernatant liquor for the mensuration of cholesterol, with the cholesterol substratum that do not connect bacterium in contrast.The mensuration of cholesterol adopts ferric ammonium sulfate colorimetry, with reference to the measuring method of GB/T5009.128-2003 Food Cholesterol, measures its OD560nm value, and the rate of decomposition of cholesterol is calculated as follows:
Utilize the Triglyceride Reagent box of Changchun Hui Li Bioisystech Co., Ltd to measure the content of triglyceride level in substratum, the triglyceride level rate of decomposition of milk-acid bacteria is calculated as follows:
Triglyceride level rate of decomposition %=(total content of triglyceride-residual value)/total content of triglyceride * 100%
156 strains of lactic acid bacteria decompose the ability of cholesterol and triglyceride level in Table 1.
Table 1 milk-acid bacteria is decomposed the ability of cholesterol and triglyceride level
| Bacterial strain | Degrading rate of cholesterol/% | Triglyceride level degradation rate/% |
| Lp2 | 51.82±5.67 | 5.33±1.65 |
| Lr5 | 61.00±1.60 | 36.72±2.72 |
| Lf7 | 60.61±4.22 | 28.71±3.53 |
| hsryfm1301 | 61.90±2.35 | 37.22±2.95 |
| L6 | 41.36±2.39 | 18.55±2.75 |
| L10 | 40.36±2.67 | 15.22±2.16 |
| L11 | 40.15±2.37 | 20.22±1.55 |
| S6 | 39.08±1.95 | 17.87±1.97 |
| L8 | 38.54±2.12 | 12.25±1.05 |
| S10 | 37.52±2.66 | 19.76±1.87 |
| L1 | 35.69±2.47 | 8.98±1.56 |
| V8 | 32.50±2.08 | 17.25±1.85 |
| S5 | 31.15±2.11 | 11.57±1.77 |
| S7 | 31.07±2.05 | 9.75±1.53 |
| V2 | 30.65±2.55 | 9.98±1.59 |
| S9 | 30.35±2.17 | 14.56±1.58 |
| L4 | 27.15±1.89 | 8.29±1.31 |
| L2 | 27.12±2.25 | 15.56±1.53 |
| S3 | 25.39±2.05 | 17.35±1.52 |
| S4 | 25.32±1.68 | 12.65±1.71 |
| L3 | 22.87±1.23 | 12.55±1.62 |
| S8 | 21.98±1.77 | 8.71±1.59 |
| S2 | 21.17±1.55 | 17.35±1.57 |
| T1 | 20.58±1.88 | 8.71±1.76 |
| T6 | 20.15±1.59 | 16.45±1.56 |
| T5 | 19.98±1.57 | 17.32±1.68 |
| L5 | 19.78±1.55 | 18.25±1.59 |
| T3 | 16.89±1.47 | 12.33±1.75 |
| L9 | 15.58±1.62 | 20.39±1.59 |
| V3 | 15.28±1.19 | 7.25±1.39 |
| All the other 126 strains | <15.00% | <5.00% |
Result shows, in separated 156 strains of lactic acid bacteria that obtain, the ability that 30 strains of lactic acid bacteria decompose cholesterol and triglyceride is stronger; Wherein cholesterol and triglyceride level degradation rate are greater than respectively 30.35% and 5.33% bacterial strain totally 16 strains; The cholesterol of bacterial strain hsryfm1301 of the present invention and the degradation rate of triglyceride level are the highest, are respectively 61.90% and 37.22%.
The mensuration of the acidproof bile tolerance ability of 5 milk-acid bacteria
The milk-acid bacteria having activated is made to bacteria suspension, get the bacterial suspension inoculation of 1.0mL to the pH3.0 simulated gastric fluid of 9.0mL, 37 ℃ of cultivations, measure viable count at 0h and 3h with colony counting method respectively, calculate its survival rate (%).
Viable count * 100% of viable count/0h of survival rate (%)=3h
The milk-acid bacteria having activated is seeded to respectively in the MRS substratum containing 0.00% (i.e. blank), 0.10%, 0.30% and 0.50% cholate by 3% inoculum size, 37 ℃ of cultivations, at 0h and 3h, with colony counting method, measure viable count respectively, calculate its survival rate (%).
Viable count * 100% of viable count/0h of survival rate (%)=3h
The acidproof bile tolerance ability of 16 strains of lactic acid bacteria is in Table 2.
The acidproof bile tolerance ability of table 2 milk-acid bacteria
Result shows, the simulated gastric fluid that 16 strains of lactic acid bacteria are 3.0 at pH and gallbladder salinity are in 0.10%, 0.30%, 0.50% MRS substratum, to cultivate 3h survival rate to be greater than respectively 12.00%, 12.50%, 10.00% and 8.00%; In the simulated gastric fluid that is wherein 3.0 at pH, survival rate is greater than 20%, and survival rate is all greater than 8.50% bacterial strain totally 8 strains in the MRS substratum of 0.10%, 0.30%, 0.50% cholate; Wherein the acidproof bile tolerance ability of bacterial strain hsryfm1301 of the present invention is stronger, and the simulated gastric fluid that is 3.0 at pH and gallbladder salinity are that in 0.10%, 0.30% and 0.50% MRS substratum, survival rate is respectively 71.43%, 66.37%, 64.55% and 19.12%.
The mensuration of 6 milk-acid bacteria bacteriostasis
Adopt Oxford agar diffusion method to detect the bacteriostasis of 8 strains of lactic acid bacteria.By pathogenic colon bacillus, Salmonellas and streptococcus aureus activation 24h, getting 0.2mL pathogenic bacterium bacterium liquid evenly coats on LB culture medium flat plate, after coating, with dull and stereotyped, without visible water droplet, be as the criterion, and with tweezers, 3 aseptic Oxford cups are put into culture dish gently, draw 0.2mL milk-acid bacteria bacteria suspension in the cup of Oxford.Plate is placed in to 3-4 ℃ of refrigerator diffusion 24h, then cultivates 24h for 37 ℃, measure antibacterial circle diameter, the results are shown in Table 3.
The biocidal property of table 3 milk-acid bacteria
By table 3, found, 8 strains of lactic acid bacteria have certain inhibition ability to pathogen enterobacteria, wherein the inhibition zone of intestinal bacteria, Salmonellas, three kinds of pathogenic bacterium of streptococcus aureus are all greater than bacterial strain totally 4 strains of 8.65mm; Bacterial strain hsryfm1301 is stronger to the inhibition ability of intestinal bacteria, Salmonellas and streptococcus aureus, and antibacterial circle diameter is respectively 14.67mm, 11.35mm and 10.67mm.
7 milk-acid bacteria antibiotic sensitivity tests
Use quick paper disk method to detect the susceptibility of strains, microbiotic and scraps of paper concentration are in Table 4.
Drawing 1.0mL bacteria suspension joins in the MRS solid medium after 15.0mL sterilizing thawing (40-50 ℃), on vortex oscillation mixing tank, be mixed evenly rear being poured into rapidly in the aseptic plate of 100mm * H20mm, shake up, after solidifying, flat board is placed with the standard drug scraps of paper, in 37 ℃ of constant incubators, be inverted and cultivate, after 48h, measure and record antibacterial circle diameter, the < < antibacterials sensitivity test operative norm > > (2010 editions) formulating according to CLSI judges that milk-acid bacteria is to antibiotic resistance, the results are shown in Table 4.
The resistance of table 4 milk-acid bacteria
Note: R is resistance, and M is medium sensitivity, and S is responsive.
By table 4, found, 4 strains of lactic acid bacteria have certain resistance to penicillin G, norfloxicin, Ciprofloxacin, and other antibiosis are have to susceptibility in various degree; Five kinds of microbiotic are all responsive usually to Kefzol, Cephradine, tsiklomitsin, the mould profit of chlorine and good fortune for bacterial strain hsryfm1301, and to Streptomycin sulphate medium sensitivity, resistance is lower than other bacterial strains.
The evaluation of 8 milk-acid bacterias
Comprehensive above test-results discovery, bacterial strain hsryfm1301 of the present invention has stronger decomposition cholesterol and triglyceride level ability, and its acidproof bile tolerance ability is also better than other bacterial strains simultaneously, and it is to the bacteriostasis of pathogen enterobacteria is strong and resistance is lower.Therefore through comparing, bacterial strain hsryfm1301 over-all properties is the most excellent.By gramstaining, find, bacterial strain hsryfm1301 of the present invention is shaped as rod-short, as Fig. 1.
(1) sugar fermentating test
According to < < uncle Jie Shi Bacteria Identification handbook > >, bacterial strain hsryfm1301 is carried out to sugar fermentating test, result is as shown in table 5.
Table 5hsryfm1301 physiological and biochemical test result
Note: "+" 90% bacterial strain is positive; "-" 90%,, bacterial strain was negative.
By table 5, found, bacterial strain hsryfm1301 of the present invention is lactobacillus rhamnosus, and it can utilize glucose, lactose, N.F,USP MANNITOL, maltose, cellobiose, melizitose, can not utilize raffinose and wood sugar.
(2) API identifies
Utilize API50CHL series indentifying substance bar to identify bacterial strain hsryfm1301, identify that figure is as Fig. 2, qualification result is in Table 6 and Fig. 3.
The API50CHL system identification result of table 6hsryfm1301
| Numbering | Fermentation tube | Bacterial strain | Numbering | Fermentation tube | Bacterial strain |
| 0 | CTRL | - | 25 | ESC | - |
| 1 | GLY | - | 26 | SAL | + |
| 2 | ERY | - | 27 | CEL | + |
| 3 | DARA | - | 28 | MAL | + |
| 4 | LARA | - | 29 | LAC | + |
| 5 | RIB | + | 30 | MEL | + |
| 6 | DXYL | - | 31 | SAC | + |
| 7 | LXYL | - | 32 | TRE | + |
| 8 | ADO | - | 33 | INU | - |
| 9 | MDX | - | 34 | MLZ | + |
| 10 | GAL | + | 35 | RAF | + |
| 11 | GLU | + | 36 | AMD | - |
| 12 | FRU | + | 37 | GLYG | - |
| 13 | MNE | + | 38 | XLT | - |
| 14 | SBE | + | 39 | GEN | + |
| 15 | RHA | - | 40 | TUR | + |
| 16 | DUL | + | 41 | LYX | - |
| 17 | INO | - | 42 | TAG | + |
| 18 | MAN | + | 43 | DFUC | + |
| 19 | SOR | + | 44 | LFUC | + |
| 20 | MDM | + | 45 | DARL | - |
| 21 | MDG | + | 46 | LARL | - |
| 22 | NAG | + | 47 | GNT | - |
| 23 | AMY | + | 48 | 2KG | - |
| 24 | ARB | + | 49 | 5KG | - |
Note: "+" represents positive; “ ?" represent negative
From table 6 and Fig. 3, bacterial strain hsryfm1301 of the present invention is lactobacillus rhamnosus, and identification rate is 99.9%.
(3) bacterial strain 16S rDNA order-checking is identified
Using bacterial strain hsryfm1301 genomic dna as the template of pcr amplification, adopt general 16S rDNA primer to carry out pcr amplification, amplification is shown in Fig. 4.After electrophoresis detection amplified production, deliver to Shanghai Sangon Biological Engineering Technology And Service Co., Ltd and check order.The sequence that order-checking is obtained (SEQ ID NO.1) compares with the sequence in GenBank database, the results are shown in Table 7.
Table 716S rDNA comparison result
| Bacterial strain | The bacterial strain that sibship is nearest | Homology |
| hsryfm1301 | Lactobacillus rhamnosus (Lactobacillus rhamnosus GG) | 99% |
In conjunction with physiological and biochemical test, API reagent strip, identify and 16S rDNA order-checking evaluation, bacterial strain hsryfm1301 of the present invention is accredited as to lactobacillus rhamnosus.
Application Example
1 making method containing strain fermentation breast of the present invention
Raw dairy is after stdn, add 6% sugar, be preheated to 60-65 ℃, 12-20Mpa pressure homogeneous, after 95 ℃/5min thermal treatment, be cooled to 42-45 ℃, by 3% inoculum size inoculation lactobacillus rhamnosus hsryfm1301 and streptococcus thermophilus grx 02 mixed culture fermentation agent, 42 ℃ of fermentation pH to 4.2-4.5, are then placed in 4 ℃ of storages.
2 making methods containing bacterial strain lactobacillus drink of the present invention
Skimmed milk powder is recovered to 12.0%(W/W) recovery skimming milk 350kg, adopt after 95 ℃/8-10min thermal treatment, be cooled to 37-40 ℃; Inoculation 3.0%(W/W) lactobacillus rhamnosus hsryfm1301 starter, in 37 ℃ of condition bottom fermentation 20-24h, terminal acidity control is at 155-170 ° of T; Add 650kg through pectin and the 13.8%-15.4%(W/W of the sugar of 90-110 ℃/5-10s sterilization, the mixture of stablizer (this mixture consist of 0.9%-1.4%(W/W)) sucrose), mix, at 20-25Mpa homogeneous; Be cooled to 15-20 ℃, adopt aseptic or health is filling, in low temperature (4-7 ℃) preservation and sale.
The making method of 3 bacterial strain powdery products of the present invention
Lactobacillus rhamnosus hsryfm1301 fermented-milk is refrigerated to-50 ℃--60 ℃, after dry, water content is 3%-4%, be crushed to Powderedly, in powdery product, add milk powder, oligose, maltodextrin etc. to make Powdered or sheet bulk product, for routine use or as medicine, healthcare products.
4 lactobacillus rhamnosus hsryfm1301 fermented-milks, lactobacillus drink and powdery product functional performance are analyzed
Get respectively the fermented-milk 0.2mL of embodiment 1 preparation, the powdery product 0.5g of the lactobacillus drink 0.6mL of embodiment 1 preparation and embodiment 3 preparations, join in MRS-THIO cholesterol substratum, cultivate after 24h for 37 ℃, get nutrient solution 0.2mL, add the vibration of 1.0mL dehydrated alcohol to mix 1min, add again the vibration of 3.8mL dehydrated alcohol to mix 1min, after standing 5min, vibration mixes again, the centrifugal 10min of 4000r/min, get 2.0mL supernatant liquor for the mensuration of cholesterol, with the cholesterol substratum that do not connect bacterium in contrast.The mensuration of cholesterol adopts ferric ammonium sulfate colorimetry, with reference to the measuring method of GB/T5009.128-2003 Food Cholesterol, measures its OD560nm value, and the rate of decomposition of cholesterol is calculated as follows:
Utilize the Triglyceride Reagent box of Changchun Hui Li Bioisystech Co., Ltd to measure the content of triglyceride level in substratum, the triglyceride level rate of decomposition of fermented-milk, lactobacillus drink and powdery product is calculated as follows:
Triglyceride level rate of decomposition %=(total content of triglyceride-residual value)/total content of triglyceride * 100%
Hsryfm1301 fermented-milk, lactobacillus drink and powdery product decompose the ability of cholesterol and triglyceride level in Table 8.
The ability that table 8hsryfm1301 fermented-milk, lactobacillus drink and powdery product decompose cholesterol and triglyceride level
| ? | Degrading rate of cholesterol/% | Triglyceride level degradation rate/% |
| Hsryfm1301 fermented-milk | 45.65±2.55 | 25.35±1.78 |
| Hsryfm1301 lactobacillus drink | 40.26±1.27 | 22.57±1.86 |
| Hsryfm1301 powdery product | 42.05±2.18 | 24.58±1.15 |
By table 8, found, hsryfm1301 fermented-milk, lactobacillus drink and powdery product all have the ability of decomposing cholesterol and triglyceride level.
The product of above strain fermentation of the present invention all has the effect of decomposing cholesterol and triglyceride level, and the lactobacillus drink that the fermented-milk of this bacterial strain and fermented-milk are made, powdery product all have the effect of decomposing cholesterol and triglyceride level.
Claims (4)
1. come from probiotic bacterium lactobacillus rhamnosus (Lactobacillus rhamnosus) hsryfm1301 for Bama of Guangxi Changshou village, it is characterized in that this bacterium has the cholesterol of decomposition and triglyceride level ability, its preserving number is CGMCC NO.8545.
2. described in claim 1, lactobacillus rhamnosus hsryfm1301 has the application in the fermented-milk that decomposes cholesterol and triglyceride level ability in preparation.
3. described in claim 1, lactobacillus rhamnosus hsryfm1301 has the application in the leben that decomposes cholesterol and triglyceride level ability in preparation.
4. described in claim 1, lactobacillus rhamnosus hsryfm1301 has the application in the active bacteria formulation that decomposes cholesterol and triglyceride level ability in preparation.
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| CN103898017B (en) * | 2014-03-28 | 2016-05-25 | 皇氏集团股份有限公司 | The application of the Lactobacillus rhamnosus that comes from Bama of Guangxi Changshou village aspect auxiliary antilipemic |
| WO2016008084A1 (en) * | 2014-07-15 | 2016-01-21 | 王国全 | Autologous lactic acid bacteria manufacturing method and system |
| CN109439778A (en) * | 2018-12-14 | 2019-03-08 | 扬州大学 | Derived from mark primer pair, identification method and its application of the Lactobacillus rhamnosus hsryfm 1301 of bar horse Changshou village |
| CN109439778B (en) * | 2018-12-14 | 2021-06-04 | 扬州大学 | Mark primer pair of lactobacillus rhamnosus hsryfm1301 derived from Bama longevity village, identification method and application thereof |
| CN109576182A (en) * | 2018-12-20 | 2019-04-05 | 江苏恒康生物科技有限公司 | A kind of strong resistance Lactobacillus rhamnosus A-4 and application thereof |
| CN110742124A (en) * | 2019-12-02 | 2020-02-04 | 扬州大学 | Method for improving comprehensive bacteriostatic ability of milk wine fermentation liquor |
| CN114711293A (en) * | 2022-04-29 | 2022-07-08 | 皇氏集团遵义乳业有限公司 | Lactobacillus rhamnosus fermented milk and preparation method thereof |
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