CN109517765A - A kind of streptococcus fecalis and its application - Google Patents
A kind of streptococcus fecalis and its application Download PDFInfo
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- CN109517765A CN109517765A CN201910030875.9A CN201910030875A CN109517765A CN 109517765 A CN109517765 A CN 109517765A CN 201910030875 A CN201910030875 A CN 201910030875A CN 109517765 A CN109517765 A CN 109517765A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The present invention provides a kind of streptococcus fecalis, in 2018 submission China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 17, (CGMCC) give preservation, deposit number are as follows: CGMCC No.16128.Streptococcus fecalis provided by the invention can generate bacteriocin, inhibit pathogen ability strong, and be made as survival rate height after freeze-dried powder.
Description
Technical field
The invention belongs to human body microecology technical field more particularly to a kind of streptococcus fecalis and its applications.
Background technique
Lactic acid bacteria (Lactic acid bactria, LAB) is that one kind can generate a large amount of creams using fermentable carbohydrate
The common name of the gram-positive bacterium of acid, streptococcus fecalis also belongs to such.There is scholar by lactic acid bacteria is defined as: cell is leather
Lan Shi is positive, cellular morphology be it is rod-shaped or spherical, catalase reaction is negative, the main lactic acid producing of fermentable carbohydrate,
Facultative or obligate anaerobe.50% or more consumption of glucose generates lactic acid, does not form endospore, without motion, or only minority
Movement.Lactic acid bacteria is extensive in distributed in nature, closely related with human lives, can perch in people and various animals enteron aisle and its
In its organ, and the category kind bacterium having causes a disease to people, and probiotic lactobacillus is generally defined as being beneficial to the bacterium of host health,
In terms of being widely used in food processing, medicines and health protection.
In terms of the classification work of current lactic acid bacteria, this kind of bacterium divides at least 23 categories now on systematic bacteriology, produces
The probiotic lactobacillus of bacteriocin generally falls into lactobacillus, enterococcus spp and Bifidobacterium.These probiotic lactobacillus are allusion quotations
The chemoorganotrophic bacterium of type, and energy fermentable carbohydrates generate lactic acid as major end products.On taxology, there is hair
Ferment same type carbohydrate has resistance to the NaCl of various concentration, can be grown on Different Nutrition medium, different restrictions
Temperature range and certain resistance is known as to antibiosis.In addition, based on them to low ph value and bile resistance and temperature
The physiological properties such as growth scope, are grown in gastrointestinal tract, it would be beneficial in lactic acid bacteria bacteriocinogeny.
The effect of probiotics is received by more and more people, and probiotics sales volume rises year by year at present, main bacteria seed
For lactobacillus acidophilus and Bifidobacterium, but the function and effect of probiotics be not it is expected as it is ideal, due to lactic acid bacteria
Tolerance is poor, not resistant against high temperatures and soda acid and some metal ions, therefore, determines that the factor of enteron aisle dominant bacteria not only depends on
In the acid producing ability of strain, but also it is related with strain whether to generate the factors such as bacteriocin.Bacteriocin is being sent out by certain bacteriums
Passing through a kind of approach to affiliation that Ribosome biogenesis mechanism generates during ferment has suppression (to kill) the active albumen of bacterium or polypeptide
Substance.Bacteriocin is a kind of selectively acting in the antibacterial material of bacterium target cell, can be by gram-positive bacteria gram
Negative bacterium or certain archeobacterias generate.Bacteriocin with no drug resistance, do not remain, be able to suppress or kill the spies such as certain pathogenic bacteria
Point, it and the entirely different property of antibiotic compensate for the defect of conventional antibiotic, it is considered to be food additives, cosmetics,
Skin care, inhibit pathogen and adjust intestinal flora good material, medicine, food, etc. fields have well apply before
Scape.Since bacteriocin itself has many advantageous properties: there is preferable selective bactericidal effect, for example, the stomach of somebody's body
In have a kind of deep and remote spiral shell door rod bacterium that can cause gastric cancer, bacteriocin plays the role of inhibition and killing to it.In addition, bacteriocin is easily by human body
Some protease (such as trypsase) in alimentary canal are degraded, and will not be accumulated in animal body and be caused adverse reaction, no pair
Effect, noresidue and free from environmental pollution without drug resistance, while also；With thermal stability, and the advantages such as acidproof, low temperature resistant storage,
Bacteriocin is widely used in food preservative, and bacteriocin is better than probiotics, but without widely used main the reason is that pure
It is big to change difficulty, it is difficult to be applied to industrialized production.As the streptococcus fecalis of one of probiotics, it is widely applied still by above-mentioned limit
System, and streptococcus fecalis be made as freeze-dried vaccine powder lyophilized technique be directly related to freeze-drying after bacterium powder survival rate, be made as being lyophilized
After bacterium powder, number of viable is lower, influences its effectiveness.
Summary of the invention
For this purpose, first technical problem to be solved by this invention, which is to provide one kind, can stablize generation Bacteriology Room, inhibit
Pathogen ability is strong and is made as the high streptococcus fecalis of survival rate after freeze-dried powder.
Second technical problem to be solved by this invention is the excrement hammer that freeze-dried vaccine powder technique in the prior art obtains
The low problem of the survival rate of bacterium bacterium powder, and then a kind of preparation method of streptococcus fecalis freeze-dried powder that Strain survival rate is high is provided.
Streptococcus fecalis of the invention, in submission on July 17th, 2018, China Committee for Culture Collection of Microorganisms is common
(CGMCC) gives preservation, deposit number at microorganism center are as follows: CGMCC No.16128.
The streptococcus fecalis, 16SrDNA sequence have the sequential structure as shown in SEQ ID NO.1.The SEQ
The sequential structure of ID NO.1 is specific as follows:
gaacgctggcggcgtgcctaatacatgcaagtagaacgctgaagagaggagcttgctcttcttggatg
agttgcgaacgggtgagtaacgcgtaggtaacctgccttgtagcgggggataactattggaaacgatagctaatac
cgcataacaatggatgacacatgtcatttatttgaaaggggcaattgctccactacaagatggacctgcgttgtat
tagctagtaggtgaggtaatggctcacctaggcgacgatacatagccgacctgagagggtgatcggccacactggg
actgagacacggcccagactcctacgggaggcagcagtagggaatcttcggcaatgggggcaaccctgaccgagca
acgccgcgtgagtgaagaaggttttcggatcgtaaagctctgttgtaagtcaagaacgggtgtgagagtggaaagt
tcacactgtgacggtagcttaccagaaagggacggctaactacgtgccagcagccgcggtaatacgtaggtcccga
gcgttgtccggatttattgggcgtaaagcgagcgcaggcggtttgataagtctgaagttaaaggctgtggctcaac
catagttcgctttggaaactgtcaaacttgagtgcagaaggggagagtggaattccatgtgtagcggtgaaatgcg
tagatatatggaggaacaccggtggcgaaagcggctctctggtctgtaactgacgctgaggctcgaaagcgtgggg
agcgaacaggattagataccctggtagtccacgccgtaaacgatgagtgctaggtgttggatcctttccgggattc
agtgccgcagctaacgcattaagcactccgcctggggagtacgaccgcaaggttgaaactcaaaggaattgacggg
ggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgaagaaccttaccaggtcttgacatcccgat
gctatttctagagatagaaagttacttcggtacatcggtgacaggtggtgcatggttgtcgtcagctcgtgtcgtg
agatgttgggttaagtcccgcaacgagcgcaacccctattgttagttgccatcattcagttgggcactctagcgag
actgccggtaataaaccggaggaaggtggggatgacgtcaaatcatcatgccccttatgacctgggctacacacgt
gctacaatggttggtacaacgagttgcgagtcggtgacggcgagctaatctcttaaagccaatctcagttcggatt
gtaggctgcaactcgcctacatgaagtcggaatcgctagtaatcgcggatcagcacgccgcggtgaatacgttccc
gggccttgtacacaccgcccgtcacaccacgagagtttgtaacacccgaagt。
The streptococcus fecalis is preparing recuperating gastrointestinal tract health and/or is improving health care product, dairy products, the drug of immunity
In application.
Preferably, application of the streptococcus fecalis in the health care product of preparation treatment and/or pre- anti-diarrhea, drug.
The streptococcus fecalis freeze-dried powder being prepared by the streptococcus fecalis.
The preparation method of streptococcus fecalis freeze-dried powder of the invention, includes the following steps:
Protective agent is added into the streptococcus fecalis, is uniformly mixed, pre-freeze at a temperature of placing it in -30~-50 DEG C
1-3h, distil 10-20h at a temperature of being subsequently placed in -10~-20 DEG C, then the 10-20h that distils at a temperature of being placed in -10~-25 DEG C,
Again in 20-30 DEG C of at a temperature of secondary distillation 1-5h, placed after 1-5h under vacuum to get streptococcus fecalis freeze-dried powder.
Preferably, the protective agent is skimmed milk power, maltose, sucrose, soluble starch, mannitol, sodium glutamate, L-
One of cysteine, gelatin are a variety of.
Preferably, the preparation method of the streptococcus fecalis freeze-dried powder, specifically comprises the following steps:
(1) streptococcus fecalis is taken, cultivates, obtains fermentation liquid；
(2) fermentation liquid that fermentation obtains in step (1) is taken, placing it in revolving speed is under 8000-12000r/min, from
Heart 10-30min collects bacterium mud；
(3) protective agent, the weight ratio of the protective agent and the bacterium mud are added into bacterium mud obtained in step (2)
For 3:1, it is uniformly mixed, pre-freeze 3h at a temperature of placing it in -40 DEG C, distil 15h at a temperature of being subsequently placed in -15 DEG C, then sets
Distil 2h at a temperature of 25 DEG C, after placing 2h under vacuum, makes the water content of the bacterium mud less than 5% to get streptococcus fecalis
Freeze-dried powder.
Preferably, the streptococcus fecalis is taken, is seeded in one grade fermemtation culture medium and trains according to the inoculum concentration of 0.5-3%
It supports, under conditions of temperature is 37 ± 3 DEG C, revolving speed is 100-180rpm, ferment 12-14h, obtains one grade fermemtation liquid；Then, it takes
The one grade fermemtation liquid, is seeded in second order fermentation culture medium according to the inoculum concentration of 5-10%, is 37 ± 3 DEG C, revolving speed in temperature
Under conditions of 100-180rpm, 4-6h is cultivated, second order fermentation liquid is obtained；The second order fermentation liquid is taken again, according to 5-12%'s
Inoculum concentration is seeded in three grade fermemtation culture medium, under conditions of temperature is 37 ± 3 DEG C, revolving speed is 150-250rpm, cultivates 14-
16h obtains three grade fermemtation liquid.
Preferably, the pH value of the one grade fermemtation culture medium, the second order fermentation culture medium, the three grade fermemtation culture medium
It is 7.0-7.4, and includes following component: 1.5 parts by weight of peptone；0.6 parts by weight of yeast extract powder；2 weight of glucose
Part；0.05 parts by weight of soluble starch；0.5 parts by weight of sodium chloride；0.05 parts by weight of L-cysteine salt；20 weight of Tomato juice
Part；0.01 parts by weight of Tween-80；0.5 parts by weight of lactoalbumin hydrolysate；2 parts by weight of agar powder.
Above-mentioned technical proposal of the invention, has the advantage that compared with prior art
(1) streptococcus fecalis of the present invention separates a large amount of streptococcus fecalis from human excrement and urine, according to inhibition pathogeny bacterium
The test such as ability, acid producing ability, digestion power, filtering out being capable of preferably digestible protein, meat slag, corn, starch, leaf, grass
Streptococcus fecalis, obtain streptococcus fecalis of the present invention, be used for human body because homologous strain colonizes well, streptococcus fecalis
It can inhibit harmful bacteria, and then safeguard the intestinal flora ecological balance, form biological barrier, inhibit invasion of the harmful bacteria to enteron aisle.Separately
Outside, the streptococcus fecalis can also promote intestines peristalsis and thallus raised growth to change infiltration by generating a large amount of short chain fatty acids
It presses thoroughly and prevents constipation.In addition, containing there are many ingredients such as enzyme, small peptide, short chain fatty acids, benefits in thallus disintegration product and metabolin
Fill the advantages such as nutritional ingredient.
Due to humans and animals for the necessary dietary intake that sustains life, after food enters alimentary canal, in addition to body itself
A large amount of intestinal flora is also needed outside the digestion of saliva, gastric juice, bile, the pancreatic juice of secretion etc. to go to decompose and digest food
Object.If the microecological balance of flora can be destroyed by largely taking antibiotic, food cannot sufficiently digest.We by pair
Grass, leaf, paper, starch, corn, meat slag, albumen carry out digestion trial, and the streptococcus fecalis filtered out not only inhibits pathogen energy
Power is strong, and digestible protein, meat slag, corn, starch, grass, in terms of have stronger function.
(2) streptococcus fecalis of the present invention, in the enterogastric diseases of people, especially diarrhea has played good prevention
And therapeutic effect, and the deficiency of antibiotic and vaccine is compensated for through a variety of ways, it is the health and acquisition safety of the mankind
Food opens new approach.
(3) streptococcus fecalis of the present invention can generate bacteriocin, to Escherichia coli, salmonella it is antibacterial straight
Diameter is larger, has stronger inhibition pathogen ability.
(4) streptococcus fecalis of the present invention, acid and bile salt tolerance is good, and the bacterial strain after the technological operations such as freeze-dried powder
Still ideal, seed the long shelf-life of activity.And the preparation method system of the streptococcus fecalis freeze-dried powder through the invention
It is standby, in preparation process, using specific protective agent, make to be made as the higher survival rate of holding of streptococcus fecalis after freeze-dried powder.
Detailed description of the invention
Fig. 1 is the bacterium solution microscopic examination result figure of streptococcus fecalis of the present invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
The streptococcus fecalis of the present embodiment, in submission on July 17th, 2018, China Committee for Culture Collection of Microorganisms is general
Logical microorganism center (CGMCC) gives preservation, and deposit number is CGMCC No.16128.
The separation screening of 1 streptococcus fecalis of embodiment
Deposit number of the invention is that the streptococcus fecalis of CGMCC No.16128 separates from the excrement of human body, and leads to
It is isolated to cross following method:
By 1 gram of human body intestinal canal excrement, loading is put in the sterilizing bottle by 30 glass marbles plus physiological saline is made 100 times of dilutions and filled
Divide after beating, remakes 10-3、10-4, successively to 10-8Dilution takes 10-4、10-5、10-6、10-7、10-8The bacterium solution of five dilutions
It is inoculated in the general agar plate of LYT, Yihong, methylene blue agar, BB (BS) Bifidobacterium selective medium, LBS (LC) cream respectively
Acidfast bacilli selective medium, EC enterococcus select base and Sb saccharomycete selects base.Every kind of culture medium connects 6 plates, after inoculation immediately
Spinning upside down culture dish spreads inoculation liquid uniformly in dish surface.Wherein make aerobic culture for 3, is observed after 24 hours.Another 3
A work is cultivated 48 hours.By visually observing and 45 DEG C of angle refraction light observations of disecting microscope.Every kind of bacterium colony is transplanted to LYT again
One bacterium colony of one bacterium colony of agar and fluid nutrient medium does anaerobism and aerobic respectively, while smear makees gram stain microscopy, point
138 bacterial strains are separated out, qualification result is carried out and belongs to 8 sections, 36 kinds of bacterium of 11 categories, wherein the separated strain filtered out is monthly
Subculture is primary, was lyophilized and saves before 5 generations.
The screening of seed bacterial strain is carried out according to following standard: (1) is without interior exotoxin, nontoxic, harmless, safe, without side-effects；
(2) may advantageously facilitate internal colony balance or prevention ecological disturbance；(3) high yield antibacterial substance, acid producing ability are strong；(4) comes
Probiotics derived from itself enteron aisle just has preferable adhesion property.It obtains having and inhibits pathogen, adjusting intestinal microecology flat
Weighing apparatus, the streptococcus fecalis for improving immunity of organisms.
The specific method is as follows for the separation and breeding of the streptococcus fecalis:
Excrement 1g is taken, is placed in the triangular flask equipped with 10ml buffered peptone water, shaken well.By 10 times of gradient dilutions,
Select each 0.1ml coating KF agar plate of 3 acceptable diluent degree, 37 DEG C of culture 48h.It is whole to select white, surface smooth bumps, edge
Together, the feature bacterium colony of diameter about 1.0~1.5mm.It passes and carries out Gram's staining after PTYG inclined-plane culture, it is blue for leather to select microscopy
Family name is positive, and circle forms oval, and the bacterial strain of single, pairs of or short catenation purifies culture.Bacterial strain after purification is given birth to
Long attribute testing, is inoculated with 6.5% sodium chloride broth culture medium and aesculin inclined-plane, after 37 DEG C of culture 48h, aesculin is selected to hydrolyze
Bacterial strain that is positive, growing on 6.5% sodium chloride broth culture medium is used as bacterial strain to be sieved.
Strain to be tested is inoculated with PTYG fluid nutrient medium, 37 DEG C of culture 48h, switching plate, picking single colonie is inoculated with respectively
In on the basis PTYG semisolid culturemedium and PTYG solid slope, 38 DEG C of culture 48h, and acidproof and bile tolerance is obtained by screening
Function admirable, and bacterial strain that is well-grown, meeting safety testing.
Wherein, the isolation medium of the streptococcus fecalis is buffered peptone water solution, specifically includes following ingredient: albumen
Peptone 10g, Na2HPO42g, glucose 5g, K2HPO42g, KCl 5g, distilled water 1000ml.The preparation method is as follows: taking 10ml slow
It rushes peptone water and is placed in 50ml triangular flask, be added a small amount of bead, it is spare after 121 DEG C of sterilizing 15min.Since streptococcus fecalis is to folded
Nitrogen sodium has tolerance, while having the characteristic for being resistant to 45 DEG C of high temperature, and the streptococcus fecalis selective medium selection is commercially available
KF culture medium carries out enterococcal separation in conjunction with high temperature culture conditions.By verifying, excrement chain can be effectively separated in this method
Coccus.
The obtained streptococcus fecalis is Gram-positive bacillus, and characteristic is as follows: streptococcus fecalis is Gram-positive ball
Bacterium, 0.5-1.0 μm of diameter, thallus is single or exists in pairs, irregular heap is also commonly formed, as shown in Figure 1.The excrement hammer
The medium bacterium colony bigger than normal of bacterium is in canescence, smooth, neat in edge.Its test stone are as follows: should be without miscellaneous bacteria, OD600 value is when transferred species
1.9-2.4。
The Morphological Identification of streptococcus fecalis is as follows: inclined-plane or semisolid deep layer are saved into strain streak inoculation PTYG plate,
37 DEG C of aerobic cultures are for 24 hours.
Specific experimental method is as follows:
Streptococcus fecalis N9024 plants of the basic bacteria is inoculated with PTYG fluid nutrient medium respectively, 37 DEG C of streptococcus fecalis quiet
Set culture 18h preparation production seed.Production seed is connected in PTYG fluid nutrient medium and passes 13 generations (cultural method is same as above), observation
Form, cultural character and the viable count of generation strains different with measurement, to determine the generation of production strain.
Test result is as follows for it:
The OD value and viable count of the streptococcus fecalis of the different passage numbers of table 1
When the above results display production seed reached for 13 generation, the streptococcus fecalis is not less than 1.0 × 109CFU/mL, energy
Enough guarantee being normally carried out for biological products production, shows that deposit number is the production of the streptococcus fecalis of CGMCC No.16128
Seed is stablized in 1-13 for vigor.The 1st generation of the streptococcus fecalis, the 10th generation, the 13rd generation production seed bacterium solution be composition
Uniform bacterial strain, morphological feature is consistent with primordial seed, and bacterium solution microscopic examination result is as shown in Figure 1, show the production of four kinds of bacterium
Seed is stablized in its interior morphological feature of 1-13 generation.
The streptococcus fecalis, 16SrDNA sequence have the sequential structure as shown in SEQ ID NO.1.The SEQ
The sequential structure of ID NO.1 is specific as follows: gaacgctggcggcgtgcctaatacatgcaagtagaacgctgaagagagga
gcttgctcttcttggatgagttgcgaacgggtgagtaacgcgtaggtaacctgccttgtagcgggggataactatt
ggaaacgatagctaataccgcataacaatggatgacacatgtcatttatttgaaaggggcaattgctccactacaa
gatggacctgcgttgtattagctagtaggtgaggtaatggctcacctaggcgacgatacatagccgacctgagagg
gtgatcggccacactgggactgagacacggcccagactcctacgggaggcagcagtagggaatcttcggcaatggg
ggcaaccctgaccgagcaacgccgcgtgagtgaagaaggttttcggatcgtaaagctctgttgtaagtcaagaacg
ggtgtgagagtggaaagttcacactgtgacggtagcttaccagaaagggacggctaactacgtgccagcagccgcg
gtaatacgtaggtcccgagcgttgtccggatttattgggcgtaaagcgagcgcaggcggtttgataagtctgaagt
taaaggctgtggctcaaccatagttcgctttggaaactgtcaaacttgagtgcagaaggggagagtggaattccat
gtgtagcggtgaaatgcgtagatatatggaggaacaccggtggcgaaagcggctctctggtctgtaactgacgctg
aggctcgaaagcgtggggagcgaacaggattagataccctggtagtccacgccgtaaacgatgagtgctaggtgtt
ggatcctttccgggattcagtgccgcagctaacgcattaagcactccgcctggggagtacgaccgcaaggttgaaa
ctcaaaggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgaagaaccttacc
aggtcttgacatcccgatgctatttctagagatagaaagttacttcggtacatcggtgacaggtggtgcatggttg
tcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccctattgttagttgccatcattca
gttgggcactctagcgagactgccggtaataaaccggaggaaggtggggatgacgtcaaatcatcatgccccttat
gacctgggctacacacgtgctacaatggttggtacaacgagttgcgagtcggtgacggcgagctaatctcttaaag
ccaatctcagttcggattgtaggctgcaactcgcctacatgaagtcggaatcgctagtaatcgcggatcagcacgc
cgcggtgaatacgttcccgggccttgtacacaccgcccgtcacaccacgagagtttgtaacacccgaagt。
The preparation of 2 streptococcus fecalis freeze-dried powder of embodiment
The streptococcus fecalis of the present embodiment is the bacterial strain that deposit number is CGMCC No.16128, is prepared via a method which
For streptococcus fecalis freeze-dried powder:
(1) streptococcus fecalis is taken, is seeded in one grade fermemtation culture medium and cultivates according to 1% inoculum concentration, in temperature
Under conditions of being 100-180rpm for 37 ± 3 DEG C, revolving speed, ferment 12-14h, obtains one grade fermemtation liquid；Then, the level-one is taken
Fermentation liquid is seeded in second order fermentation culture medium according to the inoculum concentration of 5-10%, is 37 ± 3 DEG C, revolving speed 100- in temperature
Under conditions of 180rpm, 4-6h is cultivated, second order fermentation liquid is obtained；The second order fermentation liquid is taken again, according to the inoculum concentration of 5-12%
It is seeded in three grade fermemtation culture medium, under conditions of temperature is 37 ± 3 DEG C, revolving speed is 150-250rpm, cultivates 14-16h, obtain
To three grade fermemtation liquid；
Wherein, the one grade fermemtation culture medium, the second order fermentation culture medium, the three grade fermemtation culture medium are same training
Base is supported, pH value is 7.0-7.4, and includes following component: 1.5 parts by weight of peptone；0.6 parts by weight of yeast extract powder；
2 parts by weight of glucose；0.05 parts by weight of soluble starch；0.5 parts by weight of sodium chloride；0.05 parts by weight of L-cysteine salt；West
Red 20 parts by weight of persimmon juice；0.01 parts by weight of Tween-80；0.5 parts by weight of lactoalbumin hydrolysate；2 parts by weight of agar powder.
(2) fermentation liquid that fermentation obtains in step (1) is taken, is placed it in centrifuge, revolving speed 8000r/min,
It is centrifuged 30min, collects bacterium mud；
(3) be added the protective agent into bacterium mud obtained in step (2), the protective agent be sucrose, skimmed milk power and
Gelatin is mixed according to the weight ratio of 5:3:1.The sucrose, the skimmed milk power, the gelatin are uniformly mixed, through 115
DEG C sterilizing 30min, after being cooled to room temperature, aseptically mixes according to the weight ratio of 3:1 with the bacterium mud, it is outstanding that bacterium is made
Liquid, pre-freeze 3h at a temperature of placing it in -40 DEG C, distil 15h at a temperature of being subsequently placed in -15 DEG C, then is placed in 25 DEG C of temperature
Lower distillation 2h makes the water content of the bacterium mud less than 5% to get streptococcus fecalis freeze-dried powder after placing 2h under vacuum.
The preparation of 3 streptococcus fecalis freeze-dried powder of embodiment
The streptococcus fecalis of the present embodiment is the bacterial strain that deposit number is CGMCC No.16128, is prepared via a method which
For streptococcus fecalis freeze-dried powder:
(1) streptococcus fecalis is taken, in the case where temperature is 37 DEG C, 36~48 are cultivated in the solidification cow's milk that initial pH value is 7.5
Hour, as the preferred implementation of the present embodiment, the streptococcus fecalis is seeded to by 3% inoculum concentration as the solidifying of culture medium
Gu on cow's milk, and the culture by the way of shaking flask culture；
(2) fermentation liquid that fermentation obtains in step (1) is taken, is placed it in centrifuge, revolving speed 8000r/min,
It is centrifuged 30min, collects bacterium mud；
(3) be added the protective agent into bacterium mud obtained in step (2), the protective agent be sucrose, skimmed milk power and
Gelatin is mixed according to the weight ratio of 5:3:1.The sucrose, the skimmed milk power, the gelatin are uniformly mixed, through 115
DEG C sterilizing 30min, after being cooled to room temperature, aseptically mixes according to the weight ratio of 3:1 with the bacterium mud, it is outstanding that bacterium is made
Liquid, pre-freeze 3h at a temperature of placing it in -40 DEG C, distil 15h at a temperature of being subsequently placed in -15 DEG C, then is placed in 25 DEG C of temperature
Lower distillation 2h makes the water content of the bacterium mud less than 5% to get streptococcus fecalis freeze-dried powder after placing 2h under vacuum.
The preparation of 4 streptococcus fecalis freeze-dried powder of embodiment
The streptococcus fecalis of the present embodiment is the bacterial strain that deposit number is CGMCC No.16128, is prepared via a method which
For streptococcus fecalis freeze-dried powder:
Protective agent is added into the streptococcus fecalis, is uniformly mixed, pre-freeze 1h at a temperature of placing it in -30 DEG C, so
Distil 10h at a temperature of being placed on -20 DEG C, then the 20h that distils at a temperature of being placed in -10 DEG C, then the secondary liter at a temperature of 30 DEG C
Magnificent 5h, under vacuum to get streptococcus fecalis freeze-dried powder after placement 1h.
In the present embodiment, the protective agent is skimmed milk power, be alternative implementation as the present embodiment, the protection
Agent also can be replaced skimmed milk power, maltose, sucrose, soluble starch, mannitol, sodium glutamate, L-cysteine, in gelatin
It is one or more.
Effete test embodiment
In order to verify the technical effects of the present invention, following experiment is carried out:
Experiment one, the preparation storage life test for the streptococcus fecalis seed that deposit number is CGMCC No.16128
Taking deposit number is the streptococcus fecalis of CGMCC No.16128, by basic bacteria freeze-dried powder in -80 DEG C of conditions
Lower preservation, storage life are 2 years, and every two years detection is primary.
Wherein, the method for count plate is specific as follows:
It is sterile to weigh streptococcus fecalis preparation described in 3g, 27mL fluid nutrient medium or physiological saline is added, sufficiently shakes up and (uses number
A glass marble shake beat) afterwards make 10 times be serially diluted, select acceptable diluent degree be inoculated with.Using LBS selective medium, bacterium
Each dilution be inoculated with 3 plates by 0.2mL, and rapid pivotal plate keeps bacterium solution even in media surface stream.It is small to cultivate 48
Shi Hou observes the bacterial growth situation on each plate, and counts.When the clump count on each plate is less than 10 or greater than 300
When, last dilution should be readjusted, is redeterminated.Viable count is calculated according to the following formula according to 3 plate total plate counts:
Testing result is as follows:
2 bacterial strain of table saves the viable count of different time at -80 DEG C
3 bacterial strain of table saves the viable count of different time at 4 DEG C
4 bacterial strain of table saves the viable count of different time at 25 DEG C
Streptococcus fecalis of the present invention and its fermentation liquid are placed 1 year in the refrigerator compartment of the refrigerator, and inhibition disease is done after taking out
Opportunistic pathogen test inhibits pathogen ability still very strong.
Experiment two, the acid and bile salt tolerance characteristic test of bacterial strain
In order to simulate the intracorporal gastrointestinal tract environment of animal, simulated gastric fluid and simulated intestinal fluid are prepared, is carried out external acidproof resistance to
The research of gallbladder performance.Furthermore action time is also one of the major issue be considered as when such test.Due to depositing for cholate
In the permeability for changing thallus outer membrane, so generating inhibition, killing effect to probiotics, and then the survival of probiotics is influenced.
Stomach juice-resistant test: taking deposit number is the streptococcus fecalis of CGMCC No.16128, is placed in simulated gastric fluid, 37
DEG C stationary culture 0h, 1h, 2h, 4h are measured by sampling viable count, and calculate survival rate.
Its experimental result is as follows:
The salt test of resistance to cholic acid: taking deposit number is the streptococcus fecalis of CGMCC No.16128, using simulated intestinal fluid as base
0.3% cholate is added in plinth thereto, and then the streptococcus fecalis is placed in one, in 37 DEG C of stationary cultures 0h, 1h, 2h, 4h,
Viable count is measured by sampling, and calculates survival rate.
Its experimental result is as follows:
Also commercially available three kinds of streptococcus fecalis are tested using the above method in this experiment, commercially available three kinds of streptococcus fecalis
Simulation hydrochloric acid, cholate environment in cultivate 4h after, viable bacteria rate is below 43%.It can be seen that deposit number of the invention
For the acidproof and bile tolerance function admirable of the streptococcus fecalis of CGMCC No.16128.
Experiment three inhibits pathogen capacity experimental
Taking deposit number of the invention is the streptococcus fecalis, the commercially available streptococcus fecalis of CGMCC No.16128, then is taken
There is provided pathogenic bacteria are supervised by Chinese drug: pathogenic virulent Escherichia coli 44365, pathogenic virulent salmonella C-79-14,
Pathogenic virulent salmonella C-79-20.Each bacterial strain is detected respectively to the antibacterial radius of each pathogenic bacteria, and is recorded.
Its testing result is as follows:
The influence experiment of experiment four, protective agent to freeze-dried powder technique
This experiment is investigated and is acted on protectant frozen-dried protective using survival rate as index.The system in the way of in embodiment 2
Standby streptococcus fecalis freeze-dried powder, difference are only that: protective agent is replaced with to skimmed milk power, maltose, sucrose, soluble shallow lake respectively
Streptococcus fecalis is prepared in powder, sodium glutamate, L-cysteine, gelatin, mannitol, and the streptococcus fecalis tested respectively is frozen
Dry powder saves the survival rate after 30 days at normal temperature.
It can be seen that the effect is unsatisfactory as protectant for single ingredient, and protective agent is to the streptococcus fecalis
Protecting effect differs greatly.In order to verify the stability of the technique, experimental verification three times is carried out to the scheme in embodiment 2, it is living
Bacterium rate result is respectively 93.6,92.5,95.9, error rate < 4%, it was demonstrated that result is reliable and stable.
Streptococcus fecalis freeze-dried powder is prepared in the way of in embodiment 2, difference is only that: protective agent is replaced with respectively by
Three kinds of skimmed milk power, sucrose, gelatin ingredients mix according to the weight ratio of 1:1:1, and are denoted as first group；By skimmed milk power,
Three kinds of sucrose, gelatin ingredients mix according to the weight ratio of 1:3:2, and are denoted as second group；By skimmed milk power, sucrose, gelatin
Three kinds of ingredients are mixed according to the weight ratio of 1:5:3, and are denoted as third group；Three kinds of skimmed milk power, sucrose, gelatin ingredients are pressed
According to the weight ratio mixing of 2:1:2, and it is denoted as the 4th group；By three kinds of skimmed milk power, sucrose, gelatin ingredients according to 2:3:3's
Weight ratio mixing, and it is denoted as the 5th group；Three kinds of skimmed milk power, sucrose, gelatin ingredients are mixed according to the weight ratio of 2:5:1
It closes, and is denoted as the 6th group；By skimmed milk power, sucrose, three kinds of ingredients of gelatin according to 3:1:3 weight ratio mix, and by its
It is denoted as the 7th group；Three kinds of skimmed milk power, sucrose, gelatin ingredients are mixed according to the weight ratio of 3:3:2, and are denoted as the 8th
Group；The streptococcus fecalis freeze-dried powder that embodiment 2 is prepared is denoted as the 9th group.The excrement hammer that above-mentioned each group obtains is tested respectively
Bacterium freeze-dried powder saves the survival rate after 30 days at normal temperature.
Its testing result is as follows:
Experiment five, the streptococcus fecalis are to small white mouse safety testing
The small white mouse of 20-22g is taken, cleaning grade test uses small white mouse 40, divides 4 groups, every group 10, wherein 1-3 group is given
In the active bacteria formulation of streptococcus fecalis of the present invention, wherein the streptococcus fecalis is not less than 1.0 × 107CFU, dosage period
It is 3 days, after administration, observes the clinical symptoms of all small white mouses, test result is evaluated.
After observation medicine feed during the clinical settings such as spirit, feeding, drinking-water, growth, weight gain of small white mouse, and observation test
Whether animal has adverse reaction relevant to drug, such as breathing, abnormal behavior, spirit inhibition and abnormal situation of change of discharging feces.
And the weight for recording every group of small white mouse on the 13rd day that the same day (the 1st day) and experiment terminate before administration, calculate the day of small white mouse
Increase weight.
Its experimental result is as follows:
Each group small white mouse during test is all strong to live, and spirit is good, and honey stomach, the fur colour of skin is normal, defecation urination
Color form is normal, no other abnormal clinical symptoms, without sick and dead show.
It weighs within the 13rd day, and calculates each to every group of small white mouse after the same day (the 1st day), experiment before administration
Group average daily gain.As a result it see the table below:
Find that the small white mouse Average weight increasing a day of 3 experimental groups is apparently higher than the 4th group of blank control, shows the present invention from table
The streptococcus fecalis has effect of gain to small white mouse, and it is qualified that safety examination is administered.Experiment six, the streptococcus fecalis are to dog
The improvement of intestinal bacilli illness Diarrhea Model is tested
This test, as induced drug, leads to beasle dog enteric flora disturbance, structure by being administered continuously using cefalexin
Test dog antibiotic-associated diarrhea is built, specifically comprises the following steps: to take 3.5~4 monthly ages, the regular grade of 5.0~6.5kg of weight
Test uses beasle dog 25, wherein 5, as a control group, in addition 20 in such a way that gradient increases induced drug dosage, continuously
Antibiotic induction, building test dog intestinal bacilli illness Diarrhea Model are given, antibiotic induces 15~20 days, the success of structure mould.It will
20 beasle dogs induced through antibiotic are divided into high dose group, middle dose group, low dose group, model group, to high dose group, middle dose
Amount group, low dose group give the active bacteria formulation of the streptococcus fecalis of the present invention of high, medium and low dosage respectively, and model group is not given
Medicine, dosage period are 6~8 days, after administration, observe the clinical symptoms of all dogs only.
Its experimental result is as follows:
Beasle dog weight is an index that is intuitive, easily investigating, and closely related with many factors.Compare lattice to pretherapy and post-treatment
The weight of dog is analyzed.Different groups of beasle dog weight is compared two-by-two after modeling, the average weight P of dog is tested between group
> 0.05 illustrates to influence beasle dog changes of weight using Antibiotics model little.Continuously give the streptococcus fecalis agent formulation 7
After it, model group, low, middle and high dose groups are compared into P > 0.05 two-by-two, illustrates various dose streptococcus fecalis to the body of test dog
Ghost image sound is not significant.It is puppy due to testing dog in this test, weight is still in growth state, and by many factors
It influences, therefore the index can not react true weight recovery degree, only can be used as indirect indexes.Streptococcus fecalis preparation is disorderly to flora
The influence result of random beasle dog weight is as follows:
| Average weight after modeling | Average weight after treatment | |
| Control group | 7.45±0.272 | 8.33±0.272 |
| Model group | 8.13±0.272 | 8.90±0.567 |
| Streptococcus fecalis low dose group | 7.72±0.635 | 7.98±0.862 |
| Streptococcus fecalis middle dose group | 7.44±0.533 | 8.56±1.057 |
| Streptococcus fecalis high dose group | 8.03±0.429 | 8.93±0.821 |
Wherein, unit: kg,N=2 or 5.
Before modeling, each group beasle dog is vivaciously active, and coat is flat and smooth, excrement dry forming, more normally.Modeling terminates
When, control group beasle dog diet is normal, and action is active, defecation frequency rule, excrement dry forming.The test induced through antibiotic
The dilute wet, agility of stool, which gradually occurs, in dog to be reduced, occasionally has the symptoms such as anorexia or out of strength, vomiting, is belonged to antibiotic and is excessively caused enteron aisle
Classical symptom caused by flora imbalance.
Streptococcus fecalis preparation is as follows: the influence testing result of test dog excrement state
Wherein, excrement state: -- formed stool；+ forming but moisture it is slightly more；++ sticky half congealed loose stools；+++ watery stool.
The streptococcus fecalis preparation is as shown in the table to loose stools dog therapeutic effect statistical result:
Since the microorganism in excrement accounts for the major part in enteric microorganism total amount, composition reflects entire digestive system
Flora state, and because feces collection is easy, weighs convenient, we study intestinal flora by analysis excrement.It adopts
With traditional microbiological technology (dilution plate colony counting method), Bacterial diversity in dog excrement is analyzed.Through institute
After stating streptococcus fecalis successive administration 6~7 days, control group takes 2 parts of samples, and model group takes respectively with 20 dogs of streptococcus fecalis administration group
Sample, totally 22 parts of samples, carry out conventional viable plate count.Its inspection of influence of the streptococcus fecalis to beasle dog fecal microorganism quantity
It is as follows to survey result:
Wherein, log cfu/g,N=2 or 5；In table, P < 0.01 * P < 0.05, * *.
It is continuous to after streptococcus fecalis active bacteria formulation 7 days, by dog clinical symptoms, excrement situation and bacteria detection as a result,
Various dose group is counted to the curative effect of puppy intestinal bacilli illness disease.Its testing result is as follows:
Streptococcus fecalis active bacteria formulation 2 times a day, is continuously fed 10 days with 2.0g//times, can be alleviated puppy because of enteron aisle
Symptom of diarrhea caused by dysbacteriosis promotes dog only to restore colony balance.It can be seen that streptococcus fecalis of the present invention has
Recuperating gastrointestinal tract health, improves the function of immunity.
Obviously, the above embodiments are merely examples for clarifying the description, and does not limit the embodiments.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of variation or
It changes.There is no necessity and possibility to exhaust all the enbodiments.And it is extended from this it is obvious variation or
It changes still within the protection scope of the invention.
SEQUENCE LISTING
<110>Tan Ying
<120>a kind of streptococcus fecalis and its application
<130> 2018
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1412
<212> DNA
<213>streptococcus fecalis
<400> 1
gaacgctggc ggcgtgccta atacatgcaa gtagaacgct gaagagagga gcttgctctt 60
cttggatgag ttgcgaacgg gtgagtaacg cgtaggtaac ctgccttgta gcgggggata 120
actattggaa acgatagcta ataccgcata acaatggatg acacatgtca tttatttgaa 180
aggggcaatt gctccactac aagatggacc tgcgttgtat tagctagtag gtgaggtaat 240
ggctcaccta ggcgacgata catagccgac ctgagagggt gatcggccac actgggactg 300
agacacggcc cagactccta cgggaggcag cagtagggaa tcttcggcaa tgggggcaac 360
cctgaccgag caacgccgcg tgagtgaaga aggttttcgg atcgtaaagc tctgttgtaa 420
gtcaagaacg ggtgtgagag tggaaagttc acactgtgac ggtagcttac cagaaaggga 480
cggctaacta cgtgccagca gccgcggtaa tacgtaggtc ccgagcgttg tccggattta 540
ttgggcgtaa agcgagcgca ggcggtttga taagtctgaa gttaaaggct gtggctcaac 600
catagttcgc tttggaaact gtcaaacttg agtgcagaag gggagagtgg aattccatgt 660
gtagcggtga aatgcgtaga tatatggagg aacaccggtg gcgaaagcgg ctctctggtc 720
tgtaactgac gctgaggctc gaaagcgtgg ggagcgaaca ggattagata ccctggtagt 780
ccacgccgta aacgatgagt gctaggtgtt ggatcctttc cgggattcag tgccgcagct 840
aacgcattaa gcactccgcc tggggagtac gaccgcaagg ttgaaactca aaggaattga 900
cgggggcccg cacaagcggt ggagcatgtg gtttaattcg aagcaacgcg aagaacctta 960
ccaggtcttg acatcccgat gctatttcta gagatagaaa gttacttcgg tacatcggtg 1020
acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1080
gagcgcaacc cctattgtta gttgccatca ttcagttggg cactctagcg agactgccgg 1140
taataaaccg gaggaaggtg gggatgacgt caaatcatca tgccccttat gacctgggct 1200
acacacgtgc tacaatggtt ggtacaacga gttgcgagtc ggtgacggcg agctaatctc 1260
ttaaagccaa tctcagttcg gattgtaggc tgcaactcgc ctacatgaag tcggaatcgc 1320
tagtaatcgc ggatcagcac gccgcggtga atacgttccc gggccttgta cacaccgccc 1380
gtcacaccac gagagtttgt aacacccgaa gt 1412
Claims (10)
1. a kind of streptococcus fecalis, deposit number is CGMCC NO.16128.
2. streptococcus fecalis according to claim 1, which is characterized in that the 16SrDNA sequence of the streptococcus fecalis has such as
Sequential structure shown in SEQ ID NO.1.
3. streptococcus fecalis described in claims 1 or 2 is preparing recuperating gastrointestinal tract health and/or is improving the health care of immunity
Product, dairy products, the application in drug.
4. application according to claim 3, which is characterized in that the streptococcus fecalis is in preparation treatment and/or prevention abdomen
Application in the health care product that rushes down, drug.
5. a kind of streptococcus fecalis freeze-dried powder being prepared by streptococcus fecalis of any of claims 1 or 2.
6. a kind of preparation method of streptococcus fecalis freeze-dried powder, which comprises the steps of:
Protective agent is added into streptococcus fecalis described in claims 1 or 2, is uniformly mixed, places it in -30~-50 DEG C
At a temperature of pre-freeze 1-3h, distil 10-20h at a temperature of being subsequently placed in -10~-20 DEG C, then is placed at a temperature of -10~-25 DEG C
Distil 10-20h, then in 20-30 DEG C of at a temperature of secondary distillation 1-5h, is placed after 1-5h under vacuum to get streptococcus fecalis jelly
Dry powder.
7. the preparation method of streptococcus fecalis freeze-dried powder according to claim 6, which is characterized in that the protective agent is degreasing
One of milk powder, maltose, sucrose, soluble starch, mannitol, sodium glutamate, L-cysteine, gelatin are a variety of.
8. the preparation method of streptococcus fecalis freeze-dried powder according to claim 7, which is characterized in that specifically include following step
It is rapid:
(1) streptococcus fecalis described in claims 1 or 22 is taken, cultivates, obtains fermentation liquid；
(2) fermentation liquid that fermentation obtains in step (1) is taken, placing it in revolving speed is centrifugation under 8000-12000r/min
10-30min collects bacterium mud；
(3) protective agent is added into bacterium mud obtained in step (2), the protective agent and the weight ratio of the bacterium mud are 3:
1, it is uniformly mixed, pre-freeze 3h at a temperature of placing it in -40 DEG C, distil 15h at a temperature of being subsequently placed in -15 DEG C, then is placed in 25
Distil 2h at a temperature of DEG C, after placing 2h under vacuum, makes the water content of the bacterium mud less than 5% to get streptococcus fecalis freeze-drying
Powder.
9. the preparation method of streptococcus fecalis freeze-dried powder according to claim 8, which is characterized in that step (1) is specific as follows:
Streptococcus fecalis described in claims 1 or 22 is taken, is seeded in one grade fermemtation culture medium according to the inoculum concentration of 0.5-3%
Culture, under conditions of temperature is 37 ± 3 DEG C, revolving speed is 100-180rpm, ferment 12-14h, obtains one grade fermemtation liquid；Then,
The one grade fermemtation liquid is taken, is seeded in second order fermentation culture medium according to the inoculum concentration of 5-10%, is 37 ± 3 DEG C, turns in temperature
Under conditions of speed is 100-180rpm, 4-6h is cultivated, second order fermentation liquid is obtained；The second order fermentation liquid is taken again, according to 5-12%
Inoculum concentration be seeded in three grade fermemtation culture medium, under conditions of temperature is 37 ± 3 DEG C, revolving speed is 150-250rpm, culture
14-16h obtains three grade fermemtation liquid.
10. the preparation method of streptococcus fecalis freeze-dried powder according to claim 9, which is characterized in that the one grade fermemtation training
Support base, the second order fermentation culture medium, the three grade fermemtation culture medium pH value be 7.0-7.4, and include following component:
1.5 parts by weight of peptone；0.6 parts by weight of yeast extract powder；2 parts by weight of glucose；0.05 parts by weight of soluble starch；Sodium chloride
0.5 parts by weight；0.05 parts by weight of L-cysteine salt；20 parts by weight of Tomato juice；0.01 parts by weight of Tween-80；Hydrolyze newborn egg
White 0.5 parts by weight；2 parts by weight of agar powder.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101328469A (en) * | 2008-07-09 | 2008-12-24 | 扬州大学 | Streptococcus thermophilus grx02 having alcoholic liver damage protection function, preparation and use thereof |
| CN101608201A (en) * | 2009-07-13 | 2009-12-23 | 天津科技大学 | A kind of production method of novel streptococcus thermophilus bacteriocin |
| CN101691559A (en) * | 2009-10-19 | 2010-04-07 | 李军训 | Vacuum freeze-drying method for probiotic lactobacillus |
| WO2013034974A1 (en) * | 2011-09-09 | 2013-03-14 | Giovanni Mogna | Composition comprising n-acetylcysteine and/or microencapsulated gastroprotected lysozyme in association with probiotic bacteria capable of restoring the stomach's own barrier effect which is lost during the pharmacological treatment of gastric hyperacidity |
| CN104293696A (en) * | 2014-09-16 | 2015-01-21 | 李雪平 | Enterococcus faecalis HEW-A131 and application thereof |
| US20150079040A1 (en) * | 2012-04-13 | 2015-03-19 | The University Of Manchester | Probiotic bacteria |
| CN109652334A (en) * | 2019-01-11 | 2019-04-19 | 谭瑛 | A kind of complex microbial inoculum and its preparation method and application |
-
2019
- 2019-01-11 CN CN201910030875.9A patent/CN109517765A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101328469A (en) * | 2008-07-09 | 2008-12-24 | 扬州大学 | Streptococcus thermophilus grx02 having alcoholic liver damage protection function, preparation and use thereof |
| CN101608201A (en) * | 2009-07-13 | 2009-12-23 | 天津科技大学 | A kind of production method of novel streptococcus thermophilus bacteriocin |
| CN101691559A (en) * | 2009-10-19 | 2010-04-07 | 李军训 | Vacuum freeze-drying method for probiotic lactobacillus |
| WO2013034974A1 (en) * | 2011-09-09 | 2013-03-14 | Giovanni Mogna | Composition comprising n-acetylcysteine and/or microencapsulated gastroprotected lysozyme in association with probiotic bacteria capable of restoring the stomach's own barrier effect which is lost during the pharmacological treatment of gastric hyperacidity |
| US20150079040A1 (en) * | 2012-04-13 | 2015-03-19 | The University Of Manchester | Probiotic bacteria |
| CN104293696A (en) * | 2014-09-16 | 2015-01-21 | 李雪平 | Enterococcus faecalis HEW-A131 and application thereof |
| CN109652334A (en) * | 2019-01-11 | 2019-04-19 | 谭瑛 | A kind of complex microbial inoculum and its preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| EVIVIE SMITH ETARERI: "嗜热链球菌KLDS 3.1003的基因组学、生理和益生特性研究及其应用", 《万方学位论文》 * |
| JIN-YOON KIM 等: "Probiotic properties and adsorption of Enterococcus faecalis PSCT3-7 to vermiculite", 《J VET SCI》 * |
| 白鸿 主编: "《保健食品功效成分检测方法》", 30 May 2011 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111117935A (en) * | 2020-02-10 | 2020-05-08 | 农业农村部食物与营养发展研究所 | Microbial agent for inhibiting muscle synthesis and application thereof |
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