CN109706103B - With high-adhesiveness lactobacillus plantarum RS-09 and its application - Google Patents
With high-adhesiveness lactobacillus plantarum RS-09 and its application Download PDFInfo
- Publication number
- CN109706103B CN109706103B CN201910205945.XA CN201910205945A CN109706103B CN 109706103 B CN109706103 B CN 109706103B CN 201910205945 A CN201910205945 A CN 201910205945A CN 109706103 B CN109706103 B CN 109706103B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- strain
- microbial inoculum
- serum
- drug
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The present invention relates to one plant to have high-adhesiveness lactobacillus plantarum RS-09 and its application, belongs to microorganism field.The strain isolation has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center from cider, and deposit number is CGMCC NO.17118；With stronger acidproof resistance to choline salt ability, can long-term preservation, higher to human colon cancer cell HT-29 adhesive capacity, and can be colonized in enteron aisle, wherein caecum field planting rate highest；The TNF-α content in serum can be significantly reduced, IL-10 the and IL-4 content in serum is increased, there is immunoloregulation function；There is stronger fungistatic effect to Bacterium enteritidis, there is control efficiency to Bowel inflammatory disease caused by flora imbalance, can be applied to field of functional food, not only there is actual production value, but also have very important significance to body health.
Description
Technical field
The present invention relates to microorganism field more particularly to the lactobacillus plantarums with high-adhesiveness of one plant of function admirable
(Lactobacillus plantarum) and its application.
Background technique
Lactic acid bacteria is the beneficial flora of animal gastrointestinal tract, and metabolism can produce organic acid, antibacterial peptide, biacetyl, bacteriocin etc.
A variety of natural bacteriostatic active materials have and maintain normal intestinal flora balance, the healthcare functions such as strengthen immunity.As people are strong
The raising of Kang Yishi, the requirement to healthy food also step up.Main force of the lactic acid bacteria as functional food drug, is recognized
To be a kind of natural feed addictive and food additives with broad prospect of application.
Salmonella belongs to enterobacteriaceae, is the common infecting both domestic animals and human pathogen for causing intestines problem.In China,
22.2% food poisoning is to cause to seriously endanger to publilc health by salmonellal.Currently, antibiotic intervention is husky
The conventional clinical treatment of door Salmonella disease, but antibiotic usage can enhance along with serious side effect, such as drug resistance.
The shortcomings that salmonella prevents and treats, and can not only overcome antibiotic treatment, side effect is low, and can generate and be permitted to body
The beneficial effect of more other aspects；Important type one of of the lactic acid bacteria as probiotics has pollution-free, noresidue, does not generate
The features such as drug resistance, while immunity can also be improved, increase animal to the resistance of disease；Therefore, it is replaced using lactic acid bacteria
The intestines problem of the antibiotic prevention and treatment mankind or animal has become important field of research in current food and feed industry.
Lactic acid bacteria can generate antibacterial peptide, bacteriocin, class bacterium antagonistic substance and other metabolins with bacteriostasis, such as
Organic acid, hydrogen peroxide etc., these substances cooperate with lactic acid bacteria caused by lactic acid bacteria usually to inhibit spoilage organisms in food and cause a disease
The growth of bacterium；The prerequisite that lactic acid bacteria plays prebiotic effect is can be adhered in intestinal mucosa, and can be carried out field planting, numerous
It grows；Pathogenic bacteria could cause a disease after being usually only adhered to enteron aisle locally generating toxin or invading in intestinal tissue；Therefore, newborn
Sour bacterium can occupy the attachment sites in enteron aisle first, thus the effects of generating competition exclution displacement to pathogenic bacteria；So screening is high
Adhesion lactic acid bacteria is of great significance to the research and development of probiotics with application.
Summary of the invention
In view of the above-mentioned deficiencies in the prior art, provide one plant has high-adhesiveness lactobacillus plantarum RS- to the present invention
09 bacterial strain.
The present invention is screened from apple juice fermentation liquid by strain separating, and new lactobacillus plantarum is obtained.
It is a further object to provide a lactobacillus plantarum RS-09 to prepare the medicine for inhibiting gastrointestinal tract pathogen
Application in object or food.
It is a further object to provide a lactobacillus plantarum RS-09 to prepare immunoregulation medicament or health care food
Application in product.
It is a further object to provide the method for one plant of fluorescent marker lactobacillus plantarum, trace analysis plant cream bars
Bacterium adheres to situation and field planting rate in enteron aisle.
The lactobacillus plantarum RS-09 can increase IL-10 the and IL-4 content in serum, reduce the TNF-α in serum
Content；Interleukin 10 (IL-10) is a kind of efficient anti-inflammatory factors, by blocking proinflammatory factor, chemotactic factor (CF) etc., is played
The effect of anti-inflammatory；The biological effect of interleukin-4 (IL-4), including stimulation activating B cell and T cell proliferation, CD4+T is thin
Born of the same parents are divided into II type T helper cell, play a crucial role in adjusting humoral immunity and adaptive immunity；TNF-α is a kind of inflammation
Inflammation factor is adjusting immune, inflammatory reaction, is playing a crucial role in body defenses；Therefore, lactobacillus plantarum RS-09, which has, is adjusted
Immune function.
To achieve the above object, steps are as follows for the separation screening of the lactobacillus plantarum RS-09:
Step 1 takes apple fermentated juice dilution to be coated on lactic acid bacteria selective medium MRS culture medium, 37 DEG C of anaerobism trainings
48-72h, single colonie on picking plate are supported, purifying of crossing on MRS solid medium obtains pure bacterium colony；
Step 2, by largely screening, Physiology and biochemistry and 16S identification separate lactobacillus plantarum, applicant by its
It is named as lactobacillus plantarum RS-09, which is preserved in Chinese microorganism strain preservation pipe on January 7th, 2019
Reason committee common micro-organisms center, depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number are
CGMCC NO.17118；
Step 3, lactobacillus plantarum after purification are taken the mode of freeze-drying to carry out -80 DEG C and are frozen.
The biological characteristics of the lactobacillus plantarum RS-09: the bacterial strain Gram-positive, acid producing ability is strong, has relatively strong
Acidproof resistance to choline ability.
The lactobacillus plantarum RS-09 has stronger adhesiveness and field planting rate at each position of enteron aisle, and adhesion rate is high, and energy
Stablize field planting.
The lactobacillus plantarum RS-09 has obvious bacteriostasis to Escherichia coli, salmonella.
The application of the lactobacillus plantarum RS-09, including it is used to prepare animal feed additive, it is used to prepare and adjusts livestock and poultry
The regulator of intestines problem is used to prepare antibiotics of animal substitute or is used to prepare pathogenic animal sources germ bacteriostatic agent.
The lactobacillus plantarum (Lactobacillus plantarum) RS-09 can be in pathogenic animal sources pathogen
Using.
It is compared with prior art, edible the beneficial effects of the present invention are: 1, strain isolation is from apple fermentation liquid,
It can be applied in animal feed additive and livestock and poultry cultivation, ensure that the safety of product；2, there is stronger acid producing ability,
It is resistant to the concentration of low pH and high choline salt in gastroenteric environment, is survived in enteron aisle, intestinal flora balance is adjusted, is played excellent
Prebiotic performance；3, there is stronger adhesive capacity in enteron aisle, and stablizes field planting, breeding after 48h, play prebiotic work for a long time
With；4, it can be used as animal feed additive use, it is green, safe and nontoxic, low drug resistance, without side-effects, and can be improved dynamic
The immunocompetence of object, resistance of the enhancing animal to disease；5, poultry can be applied to substitute antibiotics or part substitute antibiotics
Fowl cultivation, to reduce bacterial drug resistance caused by antibiotic is abused for a long time, environment drug residue and food safety etc.
Problem.
Detailed description of the invention
Fig. 1 is lactobacillus plantarum RS-09 Gram's staining optical microscope photograph schematic diagram of the invention.
Fig. 2 is lactobacillus plantarum RS-09 stereoscan photograph schematic diagram of the invention.
Fig. 3 is the adhesive capacity schematic diagram of lactobacillus plantarum RS-09 cell of the invention.
Fig. 4 is the negative control figure of lactobacillus plantarum RS-09 cell of the invention.
Fig. 5 is the salmonella positive control figure of lactobacillus plantarum RS-09 cell of the invention.
Fig. 6 is the repulsion lab diagram of lactobacillus plantarum RS-09 of the invention to salmonella.
Fig. 7 is the competitive assay figure of lactobacillus plantarum RS-09 of the invention to salmonella.
Fig. 8 is the displacement lab diagram of lactobacillus plantarum RS-09 of the invention to salmonella.
Specific embodiment
Principles and features of the present invention are described with reference to embodiments, the given examples are served only to explain the present invention,
It is not intended to limit the scope of the present invention.
The lactobacillus plantarum (Lactobacillus plantarum) steps are as follows for the separation screening of RS-09:
Step 1 takes apple fermentated juice dilution to be coated on lactic acid bacteria selective medium MRS culture medium, 37 DEG C of anaerobism trainings
48-72h, single colonie on picking plate are supported, purifying of crossing on MRS solid medium obtains pure bacterium colony；
Step 2, by largely screening, Physiology and biochemistry and 16S identification separate lactobacillus plantarum, applicant by its
Be named as lactobacillus plantarum (Lactobacillus plantarum) RS-09, the bacterial strain pure culture was on 01 7th, 2019
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution address: Chaoyang District, Beijing City north
The institute 3 of occasion West Road 1, deposit number are CGMCC NO.17118；
Step 3, lactobacillus plantarum after purification are taken the mode of freeze-drying to carry out -80 DEG C and are frozen.
The isolated lactobacillus plantarum (Lactobacillus plantarum) RS-09 have following biology
Characteristic:
The bacterial strain Gram-positive, acid producing ability is strong, has stronger acidproof resistance to choline ability, under the conditions of pH2.5,37
DEG C it is incubated for 2h, viable count is up to 8.23 × 108CFU/mL；Under the conditions of 0.3% gallbladder salinity, 37 DEG C of incubation 2h, viable count is up to
9.68×108CFU/mL。
The lactobacillus plantarum (Lactobacillus plantarum) RS-09 bacterial strain have at each position of enteron aisle it is relatively strong
Adhesiveness and field planting rate after feeding mouse, detect plant in different time points with cFDA-SE fluorescent marker lactobacillus plantarum
Lactobacillus colonisation and enteron aisle distribution situation in 48h, caecum part adhesion rate highest, and can stablize field planting.
The lactobacillus plantarum (Lactobacillus plantarum) RS-09 bacterial strain to Escherichia coli, salmonella have
There is obvious bacteriostasis.
The lactobacillus plantarum (Lactobacillus plantarum) RS-09 application, including be used to prepare animal feeding
Feed additives are used to prepare the regulator for adjusting livestock and poultry intestinal canal diseases, are used to prepare antibiotics of animal substitute or for making
Standby pathogenic animal sources germ bacteriostatic agent.
The lactobacillus plantarum (Lactobacillus plantarum) RS-09 can be in pathogenic animal sources pathogen
Using.
Embodiment 1
Lactobacillus plantarum (Lactobacillus plantarum) RS-09 separation, purifying and identification
Lactobacillus plantarum strain isolates and purifies: apple juice fermentation liquid is mixed, takes fermentation liquid 5mL, 10 times be serially diluted to
10-6, each diluted concentration takes 200 μ L to be coated on containing 1% CaCO3On lactic acid bacteria selective medium MRS culture medium, 37 DEG C of anaerobism
48h, single colonie on picking plate are cultivated in tank, purifying of crossing on MRS solid medium is cultivated 48h in 37 DEG C of anaerobic jars, obtained
To pure bacterium colony；Gram's staining identification is carried out to bacterium colony after purification；Bacterial strain is containing CaCO3MRS culture medium on colony diameter
1-2mm, white, protrusion, there is molten calcium circle；The bacterial strain Gram-positive, nose circle direct rod shape；Thallus Gram's staining characteristic such as Fig. 1
It is shown.Scanning electron microscope identifies that picture is as shown in Figure 2.
Lactobacillus plantarum strain screening and identification: to the bacterial strain RS-09 of acquisition using microbial identification system (Mei Liai,
VITEK 2) and 16S rDNA sequencing identified；Microflora qualification result is as shown in table 1；It is logical using the 16S delivered
With primer (27F and 1492R, primer sequence 27F:5 '-AGAGTTTGATCCTGGCTCAG-3 '；1492R:5'-
GGTTACCTTGTTACGACTT-3 ') the 16S rDNA gene order of bacterial strain RS-09 is expanded and is sequenced, PCR amplification produces
Object send Hua Da gene Co., Ltd to be sequenced, and the nucleotides sequence of the 16S rDNA of bacterial strain RS-09 is classified as the sequence 1 in sequence table；Through
The comparison of 16S rDNA gene is crossed, in GenebankLactobacillus plantarumBacterial strain compares likelihood up to 100%；
It is identified in conjunction with microflora, RS-09 is a lactobacillus plantarum, is named as lactobacillus plantarum RS-09；Lactobacillus plantarum RS-
09 16S rDNA is as shown in sequence table.
1 lactobacillus plantarum RS-09 microflora qualification result of table
| Bacterial strain | dGAL | LeuA | ELLM | PheA | ProA | PyrA | dCEL | TyrA | APPA | OPS |
| RS-09 | + | + | - | + | - | - | + | - | - | - |
| Bacterial strain | dGLU | dMNE | dMAL | SAC | ARB | NAG | BGLUi | URE | BGURi | AARAF |
| RS-09 | + | + | + | + | + | + | (+) | - | - | - |
| Bacterial strain | BGALi | AARA | AGALi | BMAN | ARG | PVATE | MTE | ESC | BdFUC | dXYL |
| RS-09 | + | - | - | - | - | - | + | + | - | - |
| Bacterial strain | BNAGi | AMANi | AIFUC | PHOS | IARA | dRIB2 | GRAM | MORPH | AERO | |
| RS-09 | - | - | - | - | - | + | + | - | - |
Note: "+" represents positive reaction；"-" represents negative reaction.
Embodiment 2
Lactobacillus plantarum (Lactobacillus plantarum) measurement of RS-09 adhesive capacity
Sterile 6 orifice plates are taken, sterile cover slips are placed in every hole, and will pass on a concentration is 1 × 105The human colon carcinoma of a/mL
Cell line HT-29 is seeded to 6 orifice plates, 5% CO2, overnight incubation under 37 DEG C of environmental conditions uses until cell monolayer, discards culture solution
DMEM solution gently washs three times, and the lactobacillus plantarum bacterium with DMEM solution suspension (being free of antibiotic) of 200 μ L is added in every hole
Suspension (5 × 107The hole CFU/) and 1800 μ L DMEM culture solutions, three holes are repeated, 37 DEG C, 5%CO are set22h is cultivated in incubator
Afterwards, supernatant is abandoned, three times, every hole is added 400 μ L0.25% pancreatin and digests 5min for sterile PBS cleaning, and 400 μ L are then added contains 10%
The DMEM solution of fetal calf serum terminates digestion；Cell sheet glass is taken, Gram's staining is carried out, randomly selects 20 views under microscope
Open country counts the total number of bacteria and cell number that adhere on the cell in the visual field, and measures adhesion index, and experimental result is as schemed
Shown in 3, RS-09 is 4.97 to human colon cancer cell line HT-29 adhesion index, has certain cell adherence ability.
Embodiment 3
Lactobacillus plantarum (Lactobacillus plantarum) DYNAMIC DISTRIBUTION of colonisation in RS-09 enteron aisle
CFDA-SE labeled plant lactobacillus RS-09: 2 × 10 are taken8The RS-09 bacterium solution 200ml of CFU/mL, at 4 DEG C,
4000rpm is centrifuged 10min, is washed 2 times with cold PBS(pH7.2), is resuspended in the final concentration of 20 μm of ol/L cFDA-SE's of 20mL
In solution, 37 DEG C are protected from light incubation 20min, and PBS is washed 3 times, remove unbonded cFDA-SE molecule, PBS is resuspended to final concentration of
2×109CFU/mL。
Enteron aisle different piece colonisation DYNAMIC DISTRIBUTION in lactobacillus plantarum RS-09 Mice Body: SPF grades of 4~5 week old is chosen
Other BALB/c mouse 54 gavages the lactobacillus plantarum suspension (2 × 10 of cFDA-SE label9CFU/mL), every 500 μ of mouse
L；5 different time points (2h/12h/24h/48h/96h) take mouse intestinal, 3 mouse of each time point after gavaging；In time
Duodenum, jejunum, ileum, caecum, colon, each intestinal segment of rectum are placed in sterile working on ice, rushed repeatedly with 3ml PBS
Each intestinal segment content is washed in 15mL centrifuge tube, with 400 mesh nylon net filters, removes tissue and particulate matter；Take filtered bacterium
Liquid centrifugation 4 DEG C, 5000rpm, is centrifuged 10min, abandons supernatant, and sterile PBS is washed three times, takes the sterile PBS of 1mL to be resuspended, flow cytometer
It is detected under 488nm exciting light, the RS-09 positive rate of each intestinal segment fluorescent marker represents RS-09 colonisation in enteron aisle.
Markup model is successfully established in lactobacillus plantarum RS-09 body, and experimental result is as shown in table 2, the results showed that plant cream
Bacillus RS-09 field planting rate with higher in enteron aisle, and can stablize after 48h and be colonized and bred.
2 flow cytomery RS-09 of table is in mouse intestinal different parts positive rate
| 2h | 12h | 24h | 48h | 96h | |
| Duodenum | 11.30% | 31.80% | 6.53% | 3.98% | 10.50% |
| Jejunum | 54.50% | 49.50% | 26.50% | 23.40% | 54.50% |
| Ileum | 77.40% | 46.30% | 36.40% | 19.20% | 40.60% |
| Caecum | 65.40% | 81.00% | 54.80% | 38.40% | 69.40% |
| Colon | 66.20% | 69.10% | 64.40% | 33.10% | 75.30% |
| Rectum | 46.50% | 71.20% | 44.50% | 30.30% | 71.50% |
Embodiment 4
Lactobacillus plantarum RS-09 is to Bacterium enteritidis Adhesion inhibiyive effect
Sterile 24 orifice plate is taken, will pass on a concentration is 1 × 105The human colon cancer cell line HT-29 of a/mL is seeded to 24
Orifice plate, 5% CO2, overnight incubation under 37 DEG C of environmental conditions, until cell monolayer；Three processing are set, by fluorescent marker method, are adopted
Lactobacillus plantarum RS-09 is studied to the Adhesion inhibiyive effect of salmonella with competition, repulsion and displacement test.
Competition experiments: HT-29 cell and lactobacillus plantarum RS-09(are unmarked), salmonella (GFP label) is 5%
CO2, 1h is incubated under 37 DEG C of environmental conditions altogether, abandons supernatant, PBS is washed three times, and the digestion of 200 μ L, 0.25% pancreatin is added in every hole
5min, the DMEM solution containing 10% fetal calf serum that 400 μ L are then added terminate digestion.
Repel test: HT-29 cell and lactobacillus plantarum RS-09(are unmarked) it is incubated for 1h altogether under 37 DEG C of environmental conditions, it abandons
Supernatant, PBS are washed 1 time, and salmonella (GFP label) is added in 5% CO2, 1h is incubated under 37 DEG C of environmental conditions altogether, abandons supernatant,
PBS is washed three times, every hole is added 200 μ L, 0.25% pancreatin and digests 5min, be then added 400 μ L containing 10% fetal calf serum
DMEM solution terminates digestion.
Displacement test: being incubated for 1h altogether under HT-29 cell and salmonella (GFP label) 37 DEG C of environmental conditions, abandons supernatant,
PBS is washed 1 time, and it is unmarked that lactobacillus plantarum RS-09(is added) in 5% CO2, 1h is incubated under 37 DEG C of environmental conditions altogether, abandons supernatant,
PBS is washed three times, every hole is added 200 μ L, 0.25% pancreatin and digests 5min, be then added 400 μ L containing 10% fetal calf serum
DMEM solution terminates digestion.
The above after treatment collects liquid, and supernatant is abandoned in centrifugation, and 0.5mL PBS is resuspended, and flow cytomery is positive
Rate, as a result as shown in Fig. 4-Fig. 8, the results showed that after lactobacillus plantarum RS-09 is added, salmonella positive rate is reduced, plant cream
Bacillus RS-09 can inhibit salmonella to adhere to enterocyte by repulsion, competition or displacement.
Embodiment 5
Lactobacillus plantarum (Lactobacillus plantarum) adjusting of the RS-09 to immune function of mice
4~5 week old BALB/C mices are chosen as animal model, are divided into two groups, every group of 5 mouse；Blank control group: from
On-test, is to terminating to feed the 500 sterile PBS of μ L；Lactobacillus plantarum group: 500 μ L bacteria suspensions (5 × 10 are gavaged9CFU/mL), even
It is continuous to gavage 10 days；After 10 days, eyeball takes blood, and every mouse takes blood into 1.5ml EP pipe, is placed at room temperature for the precipitation of 30min serum
Afterwards, 3000rpm, 4 DEG C of centrifugation 20min, takes serum to dispense, freezes in -80 DEG C, carry out subsequent ELISA experiment.
The detection of TNF-α, IL-4 and IL-10 is carried out using Shanghai Excell Biology Product Co., Ltd.'s ELISA kit,
The results are shown in Table 3；By result as can be seen that lactobacillus plantarum RS-09 can reduce inflammatory factor TNF-α factor expression,
The expression for increasing anti-inflammatory factors IL-4 and IL-10 has immunoregulatory function to animal.
3 ELISA kit of table detects immune factor in serum
| IL-10(pg/mL) | IL-4(pg/mL) | TNF-α (pg/mL) | |
| Blank control group | 121.24 | 1.75 | 281.61 |
| Lactobacillus plantarum | 135.52 | 17.43 | 114.24 |
Embodiment 6
Lactobacillus plantarum (Lactobacillus plantarum) adjusting of the RS-09 to immune function of mice
4~5 week old BALB/C mices are chosen as animal model, are divided into two groups, every group of 5 mouse；Blank control group: from
On-test, is to terminating to feed the 500 sterile PBS of μ L；Lactobacillus plantarum group: 500 μ L bacteria suspensions (5 × 10 are gavaged10CFU/mL), even
It is continuous to gavage 10 days；After 10 days, eyeball takes blood, and every mouse takes blood into 1.5ml EP pipe, is placed at room temperature for the precipitation of 30min serum
Afterwards, 3000rpm, 4 DEG C of centrifugation 20min, takes serum to dispense, freezes in -80 DEG C, carry out subsequent ELISA experiment.
The detection of TNF-α, IL-4 and IL-10 is carried out using Shanghai Excell Biology Product Co., Ltd.'s ELISA kit,
The results are shown in Table 4；By result as can be seen that lactobacillus plantarum RS-09 can reduce inflammatory factor TNF-α factor expression,
The expression for increasing anti-inflammatory factors IL-4 and IL-10 has immunoregulatory function to animal.
4 ELISA kit of table detects immune factor in serum
| IL-10(pg/mL) | IL-4(pg/mL) | TNF-α (pg/mL) | |
| Blank control group | 114.5 | 2.75 | 261.58 |
| Lactobacillus plantarum | 148.71 | 15.69 | 107.61 |
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Sequence table
<110>Ludong University
<120>there is high-adhesiveness lactobacillus plantarum RS-09 and its application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 2
<211> 1386
<212> DNA
<213>lactobacillus plantarum (Lactobacillus plantarum)
<400> 2
tgttacaaac tctcatggtg tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg 60
cggcatgctg atccgcgatt actagcgatt ccgacttcat gtaggcgagt tgcagcctac 120
aatccgaact gagaatggct ttaagagatt agcttactct cgcgagttcg caactcgttg 180
taccatccat tgtagcacgt gtgtagccca ggtcataagg ggcatgatga tttgacgtca 240
tccccacctt cctccggttt gtcaccggca gtctcaccag agtgcccaac ttaatgctgg 300
caactgataa taagggttgc gctcgttgcg ggacttaacc caacatctca cgacacgagc 360
tgacgacaac catgcaccac ctgtatccat gtccccgaag ggaacgtcta atctcttaga 420
tttgcatagt atgtcaagac ctggtaaggt tcttcgcgta gcttcgaatt aaaccacatg 480
ctccaccgct tgtgcgggcc cccgtcaatt cctttgagtt tcagccttgc ggccgtactc 540
cccaggcgga atgcttaatg cgttagctgc agcactgaag ggcggaaacc ctccaacact 600
tagcattcat cgtttacggt atggactacc agggtatcta atcctgtttg ctacccatac 660
tttcgagcct cagcgtcagt tacagaccag acagccgcct tcgccactgg tgttcttcca 720
tatatctacg catttcaccg ctacacatgg agttccactg tcctcttctg cactcaagtt 780
tcccagtttc cgatgcactt cttcggttga gccgaaggct ttcacatcag acttaaaaaa 840
ccgcctgcgc tcgctttacg cccaataaat ccggacaacg cttgccacct acgtattacc 900
gcggctgctg gcacgtagtt agccgtggct ttctggttaa ataccgtcaa tacctgaaca 960
gttactctca gatatgttct tctttaacaa cagagtttta cgagccgaaa cccttcttca 1020
ctcacgcggc gttgctccat cagactttcg tccattgtgg aagattccct actgctgcct 1080
cccgtaggag tttgggccgt gtctcagtcc caatgtggcc gattaccctc tcaggtcggc 1140
tacgtatcat tgccatggtg agccgttacc ccaccatcta gctaatacgc cgcgggacca 1200
tccaaaagtg atagccgaag ccatctttca agctcggacc atgcggtcca agttgttatg 1260
cggtattagc atctgtttcc aggtgttatc ccccgcttct gggcaggttt cccacgtgtt 1320
actcaccagt tcgccactca ctcaaatgta aatcatgatg caagcaccaa tcaataccag 1380
agttcg 1386
Claims (11)
1. a kind of lactobacillus plantarum strain, which is characterized in that the bacterial strain is to be screened from apple juice fermentation liquid by strain separating
, be named as lactobacillus plantarum (Lactobacillus plantarum) RS-09, the bacterial strain is in preservation on January 7 in 2019
In China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC NO.17118.
2. lactobacillus plantarum strain according to claim 1, which is characterized in that the bacterial strain can increase the IL-10 in serum
And IL-4 content, the TNF-α content in serum is reduced, Salmonella enteritidis is reduced and is colonized quantity.
3. a kind of microbial inoculum, which is characterized in that contain lactobacillus plantarum strain as claimed in claim 2 in active constituent.
4. microbial inoculum according to claim 3, which is characterized in that the concentration of the lactobacillus plantarum strain is 108CFU/g or more.
5. microbial inoculum according to claim 4, which is characterized in that the concentration of the lactobacillus plantarum strain is 109-1010CFU/
g。
6. the microbial inoculum according to any one of claim 3-5, which is characterized in that drug is made in the microbial inoculum, drug
Dosage form is selected from least one of tablet, granule, powder, capsule, solution, suspending agent or emulsion.
7. the microbial inoculum according to any one of claim 3-5, which is characterized in that drug is made in the microbial inoculum, and drug is adopted
It is prepared with freeze drying process.
8. a kind of composition, which is characterized in that contain lactobacillus plantarum strain as claimed in claim 2 in ingredient.
9. a kind of composition as claimed in claim 8 is preparing the application in food.
10. a kind of application of composition as claimed in claim 8 in the drug that preparation inhibits gastrointestinal tract pathogen.
11. a kind of composition as claimed in claim 8 is preparing the application in immunoregulation medicament.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910205945.XA CN109706103B (en) | 2019-03-19 | 2019-03-19 | With high-adhesiveness lactobacillus plantarum RS-09 and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910205945.XA CN109706103B (en) | 2019-03-19 | 2019-03-19 | With high-adhesiveness lactobacillus plantarum RS-09 and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109706103A CN109706103A (en) | 2019-05-03 |
| CN109706103B true CN109706103B (en) | 2019-08-06 |
Family
ID=66266966
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910205945.XA Active CN109706103B (en) | 2019-03-19 | 2019-03-19 | With high-adhesiveness lactobacillus plantarum RS-09 and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109706103B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20210084917A1 (en) * | 2019-09-20 | 2021-03-25 | Siobhan Reilly | Freeze-Drying Methods |
| CN113604410B (en) * | 2021-09-18 | 2023-06-02 | 兰州大学 | Lactobacillus plantarum YT013 and application thereof |
| CN114540236B (en) * | 2022-03-07 | 2023-08-15 | 鲁东大学 | Lactobacillus fermentum and application thereof |
| CN115948285A (en) * | 2022-11-23 | 2023-04-11 | 鲁东大学 | Biological disinfectant for lactobacillus salivarius BMC-06 and livestock breeding and application |
| CN116694503B (en) * | 2023-03-27 | 2024-01-05 | 上海华聿康生物科技有限公司 | Lactobacillus plantarum Lp-HZ55 with bowel relaxing and immunity improving functions |
| CN116463264B (en) * | 2023-04-21 | 2024-02-06 | 四川大学 | Lactobacillus plantarum with colon cancer cell growth inhibition effect and application thereof |
| CN117159598B (en) * | 2023-11-02 | 2024-01-02 | 微康益生菌(苏州)股份有限公司 | Application of lactobacillus plantarum Lp18 in preparation of immunity-enhancing medicines or health-care foods and products |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103509734B (en) * | 2013-06-04 | 2016-03-30 | 鲁东大学 | A kind of Novel fruit wine biological-deacidification bacterial strain, preparation method and application thereof |
| CN108570436B (en) * | 2017-12-14 | 2020-08-04 | 浙江大学 | Lactobacillus plantarum ZJUF T17 and application thereof |
| CN109234181B (en) * | 2017-12-22 | 2020-06-09 | 浙江大学 | Lactobacillus plantarum ZJUF HN9 and application thereof |
-
2019
- 2019-03-19 CN CN201910205945.XA patent/CN109706103B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN109706103A (en) | 2019-05-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109706103B (en) | With high-adhesiveness lactobacillus plantarum RS-09 and its application | |
| CN104232515B (en) | A kind of animal bifidobacteria and its application | |
| WO2022110727A1 (en) | Application of lactobacillus paracasei et-22 in improving intestinal bacterial infection resistance and intestinal immunity | |
| CN104388345B (en) | One plant of Pedicoccus acidilacticii strain and application thereof | |
| CN114774313B (en) | Use of lactobacillus rhamnosus LRa05 in preparing constipation relieving product or intestinal flora regulating product | |
| CN109182166A (en) | One plant has effects that the Lactobacillus rhamnosus of relief of constipation and its application | |
| CN113913322B (en) | Application of bifidobacterium lactis BLA80 in relieving diarrhea and improving intestinal immunity | |
| CN104805040B (en) | A kind of bacillus subtilis formulation and preparation method and application | |
| CN114381411B (en) | Lactococcus lactis JYLL-60 and application thereof in preparation of product for improving immunity | |
| CN115094012B (en) | Preparation method and application of bacillus coagulans BC-HYC strain microbial inoculum | |
| CN108004179A (en) | The lactobacillus reuteri and application of one plant of tool probiotic properties and antagonism production enterotoxigenic Escherichia coli | |
| CN113040390B (en) | Probiotic salt-tolerant lactobacillus johnsonii and application thereof in preventing and treating pathogenic bacteria in livestock and poultry aquiculture | |
| CN109628346A (en) | Lactobacillus fermenti CQPC04 and its application in the food that preparation improves constipation | |
| WO2019205422A1 (en) | Bacillus coagulans xp and application thereof in animal feed production | |
| CN102008110B (en) | Multi-strain microbial composite beverage preparation and preparation method thereof | |
| CN106721523B (en) | Micro-ecological preparation applied to turbot culture | |
| CN113652373A (en) | Bacillus and application thereof | |
| CN113621538A (en) | Lactobacillus amylovorus and application thereof | |
| CN114990030B (en) | Lactobacillus acidophilus LA18 and application thereof in preparing product for regulating intestinal flora or immunoregulation | |
| CN115505546B (en) | Salt-tolerant bacillus for inhibiting aeromonas hydrophila and application thereof | |
| CN116286458A (en) | Lactobacillus plantarum LZ010 capable of reducing blood pressure and blood fat and application thereof | |
| CN114806953A (en) | Lactobacillus gasseri with characteristic of improving type 1 diabetes | |
| CN113930367A (en) | Lactic acid bacteria with cholesterol-reducing performance and application thereof | |
| CN113712207B (en) | Probiotic prebiotic composition for improving gastrointestinal tract immunity and application thereof | |
| CN117305187B (en) | Pediococcus acidilactici for improving intestinal health condition and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Lactobacillus plantarum rs-09 with high adhesion and its application Effective date of registration: 20211216 Granted publication date: 20190806 Pledgee: Yantai financing guarantee Group Co.,Ltd. Pledgor: LUDONG University Registration number: Y2021980015152 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20220317 Granted publication date: 20190806 Pledgee: Yantai financing guarantee Group Co.,Ltd. Pledgor: LUDONG University Registration number: Y2021980015152 |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221021 Address after: No.1666, East Ring Road, juancheng County, Heze City, Shandong Province 274600 Patentee after: SHANDONG YANGCHENG BIOTECH Co.,Ltd. Address before: 264001 No.184, Hongqi Middle Road, Zhifu District, Yantai City, Shandong Province Patentee before: LUDONG University |