CN108373984A - A kind of Lactobacillus paracasei and its application - Google Patents
A kind of Lactobacillus paracasei and its application Download PDFInfo
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- CN108373984A CN108373984A CN201810311076.4A CN201810311076A CN108373984A CN 108373984 A CN108373984 A CN 108373984A CN 201810311076 A CN201810311076 A CN 201810311076A CN 108373984 A CN108373984 A CN 108373984A
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- C—CHEMISTRY; METALLURGY
- 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
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/24—Lactobacillus brevis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
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- C—CHEMISTRY; METALLURGY
- 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
Abstract
A kind of Lactobacillus paracasei and its application, Lactobacillus paracasei (Lactobacillus paracasei) L267 provided by the invention, preserving number are：CGMCC No.14868, come from the ensilage of spontaneous fermentation, belong to microorganism field.Lactobacillus paracasei L267 has the ability of confrontation cholate and gastric acid, has the ability of production protease, has inhibiting effect to Escherichia coli flora.More immunity-regulating reacts, suitable for the preparation of the products such as food, health products, drug, food supplement.Lactic bacteria composition can be applied in feeder for fat pig.After being fed to growing and fattening pigs, to reduce feedstuff-meat ratio and the death rate, reduction delivers number of days for sale, reduces the use of antibiotic.
Description
Technical field
The present invention relates to microorganism fields, and in particular to a kind of Lactobacillus paracasei and its application.
Background technology
It is lived in body beneficial to host health at present it is known that Bacillus acidi lactici (Lactobacillus spp.) is a group
Microorganism, it safeguards that health and the effect of adjusting immune function are widely recognized as.Lactobacillus paracasei can promote body
The balance and stimulation specificity of microorganism species and enzyme and nonspecific immunologic mechanism, but when thalline is acidproof, bile tolerance energy
Enteron aisle is not entered when power deficiency, its effect will be lost.
Present rhythm of life is very fast, and people's usually can all select to take effect antibiotic of speed improves human body or poultry
The immunity of product.But the frequent change used so that the digestive tract environment in human body or animal body occurs of antibiotics
Change, capacity of self-regulation difference and microorganism species it is unbalance, for a long time using only will appear infection and more potential hazards.Cause
This, in nutrient research and domestic animals breeding, people are more desirable to greatly develop the microniological proudcts harmless to the mankind and domestic animals.
Invention content
The technical problem to be solved in the present invention is to overcome the shortcomings of the prior art, provides a kind of extraction process letter
Single, stability is high, has excellent acid producing ability, has preferable tolerance to acid, cholate, has stronger inhibition to coliform
The Lactobacillus paracasei of effect, and a kind of composition prepared using the Lactobacillus paracasei and its application are provided accordingly.
One aspect of the present invention provides a kind of Lactobacillus paracasei, Lactobacillus paracasei (Lactobacillus
Paracasei) L267, preserving number are：CGMCC No.14868, the deposit date is on November 08th, 2017, depositary institutions：China
Microorganism fungus kind administration committee common micro-organisms center.
Further, the Lactobacillus paracasei L267 has tolerance to hydrochloric acid in gastric juice and cholate.
Further, the Lactobacillus paracasei L267 has the ability of production protease.
Further, the Lactobacillus paracasei L267 has inhibiting effect to Escherichia coli flora.Further, pheS sequences
As shown in SEQ ID NO.1,16SrRNA sequences are as shown in SEQ ID NO.2.
Another aspect of the present invention provides a kind of preparation method of Escherichia coli antibacterial liquid, is placed on the secondary cheese of low temperature refrigerator
Lactobacillus L2 67, which is put into 40~50 DEG C of water-baths, to thaw, and is aseptically inoculated into 9# fluid nutrient mediums, 37 DEG C of constant temperature trainings
15h is supported, activated for 2~3 generations；The method that activated Lactobacillus paracasei L267 zymotic fluids are used low-temperature centrifugation, prepares supernatant
Liquid obtains Escherichia coli antibacterial liquid.
The present invention also provides a kind of food, are prepared with the Lactobacillus paracasei of CGMCC No.14868.
The present invention also provides a kind of lactic bacteria compositions, including deposit number is the secondary cheese breast bar of CGMCC No.14868
Bacterium and acceptable auxiliary material.
Further, further include in the lactic bacteria composition：Lactobacillus acidophilus, bifidobacterium animalis subspecies and cheese breast bar
Bacterium；The additive amount of each component is：Lactobacillus paracasei：Lactobacillus acidophilus：Bifidobacterium animalis subspecies：Lactobacillus casei=2
~6：0.5~1.5：0.5~1.5：0.5~1.
Further, auxiliary material be selected from lactose, dry starch, dextrin, cyclodextrin, sodium carboxymethyl starch, carboxy starch propionic ester,
Microcrystalline cellulose, carboxymethyl cellulose, maltodextrin, magnesium stearate.
The present invention also provides a kind of applications using above-mentioned lactic bacteria composition in feeder for fat pig.It can be with it to educating
Hog is fed, and to reduce feedstuff-meat ratio and the death rate, reduction delivers number of days for sale, reduces the use of antibiotic.
As same technical concept, the present invention also provides a kind of above-mentioned Lactobacillus paracaseis in Escherichia coli antibacterial liquid, food
Application in product, food supplement, drug, health products.
Beneficial effects of the present invention are embodied in：
1, a kind of Lactobacillus paracasei provided by the invention, the selection and breeding from the ensilage of spontaneous fermentation are identified as pair
Lactobacillus casei；The obtained Lactobacillus paracasei invented above has a good tolerance to human body gastrointestinal environment, and can be with
Production protease helps digest, and fermentation has higher viable bacteria amount, reduces fermentation costs.
2, a kind of application of the lactic bacteria composition provided by the invention in growing and fattening pigs in feed, can with its to growing and fattening pigs into
Row feeding, to reduce feedstuff-meat ratio and the death rate, reduction delivers number of days for sale, reduces the use of antibiotic.
3, Lactobacillus paracasei L267 provided by the invention fights the ability of cholate and gastric acid, and more immunity-regulating is anti-
It answers, suitable for the preparation of the products such as food, health products, drug, food supplement.
Biological deposits explanation
Classification And Nomenclature：Lactobacillus paracasei (Lactobacillus paracasei) L267 was protected on November 08th, 2017
China Committee for Culture Collection of Microorganisms's common micro-organisms center is ensconced, address is BeiChen West Road, Chaoyang District, BeiJing City 1
Institute 3, Institute of Microorganism, Academia Sinica, deposit number are：CGMCC No.14868.
Description of the drawings
The present invention will be further described in detail below in conjunction with the accompanying drawings.
Fig. 1 is the change curve of L267 pH during the fermentation.
Fig. 2 is L267 thalli morphology schematic diagrames.
Specific implementation mode
The invention discloses a kind of Lactobacillus paracasei and its application, those skilled in the art can use for reference present disclosure,
It is suitably modified technological parameter realization.In particular, it should be pointed out that all similar substitutions and modifications carry out those skilled in the art
Say it is it will be apparent that they are considered as including within the present invention.Lactobacillus paracasei of the present invention is by preferable
Embodiment is described, related personnel obviously can not depart from the content of present invention, in spirit and scope to side as described herein
Method and application are modified or suitably change and combine, to realize and apply the technology of the present invention.
One plant of Lactobacillus paracasei L267 provided by the invention, comes from the ensilage of spontaneous fermentation.Secondary cheese breast
The physio-biochemical characteristics of bacillus L267 are as shown in table 1 below：
1 physio-biochemical characteristics of table
Thalli morphology is as shown in Figure 2.
Inventor for the rate of producing acid of Lactobacillus paracasei L267, acidproof, bile tolerance, Escherichia coli Inhibition test,
Following experiment has been done in prolease activity determination experiment and growing and fattening pigs feeding in metabolite：
1. rate of producing acid-high density fermentation of Lactobacillus paracasei L267
The prebiotic effect of microniological proudcts is played, the digestion process of human intestines and stomach must be resistant to first, gastric juice can be resisted
The effect that it is killed with bile.Report time for emptying under one's belt of food such as Berrada is 90min, stomach inner pH value 3.0,
Minimum reachable 1.3, human small intestine's bile fluid concentration is between 0.03%~0.3%.
1.1 experiment material
Lactobacillus paracasei (Lactobacillus paracasei) this laboratory separating and preserving, number L267.Used
Fermentation medium and feed-batch culture based formulas are as follows：
Fermentation medium (9# culture mediums)：Soy peptone 5.0g~20.0g, beef extract 5.0g~20.0g, yeast extract
5.0g~20.0g, tryptone 5.0g~20.0g, casein peptone 5.0g~20.0g, 10% tomato juice 10ml~100ml,
K2HPO41.0g~5.0g, lemon acid diamine 1.0g~5.0g, sodium acetate 1.0g~5.0g, glucose 5.0g~20.0g, malt
Sugared 5.0g~20.0g, oligofructose 5.0g~20g, 80 0.5ml of Tween~2.0ml, MgSO4·7H2O 0.5g~1.0g,
MnSO4·4H2O 0.1g~1.0g are settled to 1L with distilled water, adjust 5~8,100 DEG C~125 DEG C sterilizing 5min of pH~
30min。
Supplemented medium：Glucose 5%~10%, maltose 5%~10%, oligofructose 5%~10%, 10% tomato
Juice 2%~5%, oligofructose 5%~10%, soy peptone 5%~10%, beef extract 5%~10%, yeast extract 5%~
10%, tryptone 5%~10%, casein peptone 5%~10%, dibasic ammonium citrate 2%~5% prepare 300ml~800ml,
Adjust 5~8,100 DEG C~125 DEG C sterilizing 5min~30min of pH.
1.2 test method
1.2.1 cultural method
1.2.1.1 glycerine seed activation；Glycerol stocks strain is taken to be seeded in 9# liquid test tubes, 37 DEG C of stationary cultures.
1.2.1.2 seed culture；By 2% inoculum concentration, appropriate amount is taken from activation bacterium solution, access is equipped with 500ml 9# cultures
In the triangular flask of the 1000ml of base, 37 DEG C of stationary cultures.
1.2.1.3 fermented and cultured；By 2% inoculum concentration, access is equipped in the fermentation tank of the 30L of 25L 9# culture mediums, and 37
DEG C, rotating speed 50r/min culture.
1.2.1.4 feed supplement tune acid；When certain fermented incubation time (pH less than most just when), feed supplement, and toward fermentation
In liquid plus alkali, adjusting pH to 6.5 continue to cultivate.
1.2.1.5 go out fermentation；When strain growth to logarithmic phase later stage or balance period early period, by tank temperature drop to 20 DEG C of left sides
The right side, and add alkali to be adjusted to 6.5 pH value.
1.2.1.6 freeze-drying；It is -4 DEG C, 4000r/min to adjust centrifuge parameters, centrifuges 10min, abandons supernatant, will receive
The bacterium mud of collection is put into freeze drier after being mixed well with protective agent and is lyophilized.
1.2.2 assay method
Cell concentration is detected using dilution pour plate counting method.
1.3 results and analysis
1.3.1 incubation pH value situation of change is as shown in table 2.
Table 2：1. the variation of Lactobacillus paracasei L267 pH during the fermentation
| Time | pH |
| 0h | 6.86 |
| 2h | 6.73 |
| 3.5h | 6.47 |
| 5h | 5.98 |
| 6h | 5.74 |
| 7h | 5.39 |
| 7.5h | 6.5 |
| 10h | 5.02 |
| 14h | 4.61 |
| 15h | 4.62 |
PH change schematic diagrams are referring to attached drawing 1.
1.3.2 each group different periods viable count is as shown in table 3.
Table 3 is freeze-dried front and back viable count control
Thalline is freeze-dried front and back feed supplement compared with no-feed supplement, viable count increases by being collected by centrifugation after fermentation；It is logical
Thalli morphology is preferable after crossing microscopy feed supplement, uniform coloring.
2. the tolerance of pair acid and cholate
2.1 experiment material
Lactobacillus paracasei (Lactobacillus paracasei) this laboratory separating and preserving, number L267.
9# culture mediums.
2.2 test method
2.2.1 cholate tolerance test
2.2.1.1 Spawn incubation；Strain is cultivated into 15h for 37 DEG C in the 9# culture mediums of sterilizing；
2.2.1.2 cholate is resistant to；Activated strains culture solution is accessed into the 9# liquid containing different gallbladder salinities with 2% inoculum concentration
In culture medium (gallbladder salinity 0.2%, 0.3%, 0.5%, 1.0%, 1.6%), while being with the 9# culture solutions without cholate
Control.0h, 2h, 4h, 6h, for 24 hours are cultivated at 37 DEG C.
2.2.1.3 plate count；Cell concentration is detected using dilution pour plate counting method.
2.2.2 sour tolerance test
2.2.2.1 Spawn incubation；Strain is cultivated into 15h for 37 DEG C in the 9# culture mediums of sterilizing.
2.2.2.2 acid tolerance；Activated strains culture solution is accessed into the 9# fluid nutrient mediums containing different pH value with 2% inoculum concentration
In (pH value is respectively 1.3,2.0,3.0), while using pH value be 6.5 9# culture solutions as control.At 37 DEG C cultivate 0min,
30min、60min、90min、120min。
2.2.2.3 plate count；Cell concentration is detected using dilution pour plate counting method.
2.3 results and analysis
2.3.1 bile tolerance result
Lactobacillus paracasei L267 is as shown in table 4 to the tolerance of different gallbladder salinities：
Table 4：Bile tolerance result
It can be seen that：Salinity is less than or equal to act on its unrestraint when 0.3%, and viable count is continuously increased, 0.5%~
Growth has certain tolerance in 1.6% gallbladder salinity.
2.3.2 acidproof result
Viable count variations of the Lactobacillus paracasei L267 in different acid culture solutions is as shown in table 5.
Table 5：Acidproof result
As can be seen that under conditions of pH=2.0, culture illustrates that the bacterial strain has acid in 2 hours still with the presence of a large amount of viable bacterias
Very strong tolerance.
One plant of selection and breeding can be with from the ensilage of spontaneous fermentation for the physiological condition of human body and animal stomach by the present invention
It is resistant to the Lactobacillus paracasei of human body and animal hydrochloric acid in gastric juice and gallbladder salinity, so as to be colonized in human body and animal body, is played
Prebiotic effect.Meanwhile the bacterial strain has good inhibiting effect to coliform, can play and adjust intestinal microflora function.
3. the inhibition of pair Escherichia coli
3.1 experiment material
Lactobacillus paracasei (Lactobacillus paracasei) this laboratory separating and preserving, number L267.Large intestine bar
Bacterium (comes from this laboratory), nutrient broth medium, 9# culture mediums.
3.2 experimental method
3.2.1 actication of culture；Glycerol stocks strain is taken to be seeded in 9# liquid test tubes, 37 DEG C of stationary culture 15h, activation 2
~3 generations.
3.2.2 the activation of Escherichia coli；The Escherichia coli being placed in refrigerator are transferred to liquid battalion according to 2% inoculum concentration
It supports in broth bouillon, 37 DEG C of constant temperature incubation 12h, activated for two generations.
3.2.3 being coated with nutrient agar plate；Activated Escherichia coli are uniformly applied to nutrient agar with rubbing method
On tablet, it is desirable that viable bacteria amount when coating is in 106cfu/ml.
3.2.4 Odontothrips loti；The nutrient agar panel for the indicator bacteria completed is spontaneously dried into 30min, then in aseptic condition
It is lower to be uniformly positioned over Oxford cup (internal diameter 6mm) on tablet, it slightly pushes, makes itself and plating medium contact surface tight,
Then 0.2ml fermented supernatant fluids are drawn in cup, 37 DEG C of constant temperature incubations for 24 hours, observe the size of inhibition zone, and it is straight to measure inhibition zone
Diameter.Each sample do three it is parallel, results are averaged.Control group adds 9# fluid nutrient medium 0.2ml in Oxford cup, culture
Condition is identical.
3.3 experimental result
Experimental result is shown：Most of lactobacillus has inhibiting effect, and L267 pairs of Lactobacillus paracasei to coliform
The inhibition of Escherichia coli is particularly evident, and inhibition zone reaches 20mm, is much larger than other kind of lactic acid bacteria, and concrete outcome see the table below shown in 6：
6 Lactobacillus paracasei L267 of table, lactic acid bacteria and Escherichia coli antagonistic effect
| Strain name | Antibacterial circle diameter (mm) |
| Lactobacillus paracasei L267 | 20 |
| Bifidobacterium breve YDB08 | 16 |
| Bifidobacterium adolescentis YDB121 | 16 |
| Bifidobacterium adolescentis YDB155 | 13 |
| Bifidobacterium bifidum YDB129 | 14 |
| Lactobacillus casei YDL194 | 10 |
| Lactobacillus acidophilus YDL211 | 13 |
| Lactobacillus acidophilus YDL126 | 7 |
| Lactobacillus acidophilus YDL165 | 16 |
| Lactobacillus bulgaricus YDL177 | 10 |
| Lactobacillus acidophilus YDL194 | 15 |
4. prolease activity measures in metabolite
4.1 experiment material
Strain name：Lactobacillus paracasei (Lactobacillus paracasei) this laboratory separating and preserving, number
L267.9# culture mediums, tyrosine, sodium carbonate, forint phenol reagent, casein, trichloroacetic acid.
4.2 experimental method
4.2.1 the formulation of tyrosine standard curve；
Prepare various concentration tyrosine standard solution (0,20,40,60,80,100ug/mL), take various concentration respectively
Tyrosine standard solution 1.00ml respectively adds sodium carbonate liquor 5.00ml, the forint dilution 1.00ml of 0.4mol/ml, is placed in 40
Develop the color 20min in DEG C water-bath, takes out, and it (is sky with the pipe without tyrosine to measure its absorbance OD660 respectively with spectrophotometer
In vain).The tyrosine standard solution of various concentration and the variation relation of OD660 are shown in Table 7.
Table 7：The tyrosine standard solution and OD of various concentration660Variation relation
| Tyrosine concentration (ug/ml) | 10 | 20 | 30 | 40 | 50 | 60 | 80 |
| OD660 | 0.101 | 0.21 | 0.308 | 0.409 | 0.513 | 0.625 | 0.81 |
With OD660Value is abscissa, draws standard curve with a concentration of ordinate of tyrosine, obtains calibration curve equation：y
=98.22x-0.3288 (R2=0.9994)
4.2.2 prepared by enzyme solution；By the Lactobacillus paracasei L267 zymotic fluids after 37 DEG C of ferment at constant temperature 18h, left in 3500
The heart 20 minutes, it is crude enzyme liquid to take supernatant.
4.2.3 casein preheats；2% casein is put into 40 DEG C of constant temperature waters, is preheated 5 minutes.
4.2.4 enzyme digestion reaction；1ml crude enzyme liquids are taken, adds the casein 1ml after preheating, shakes up, and accurately kept the temperature at 40 DEG C
10 minutes.
4.2.5 reaction is terminated；Add 0.4mol/L trichloroacetic acid 2ml, after shaking up continue waters place 20 minutes it is residual to precipitate
Albumen is stayed, 3500 leave the heart 10 minutes.Supernatant 1ml is taken, 0.4mol/L sodium carbonate liquor 5ml and 1ml forint is added at 40 DEG C
Phenol reagent keeps the temperature 20 minutes.
4.2.6 colorimetric；The colorimetric under 660nm wavelength surveys its absorbance.
4.2.7 blank control；In blank assay, add casein opposite with the sequence of trichloroacetic acid.
4.2.8 calculation formula
Proteinase activity power (U/ml)=(OD samples-OD blank) × K × 4/10 × N
Wherein, K=102.91
4 --- 4ml reaction solutions take 1ml to measure；
N --- enzyme solution extension rate；
10 --- reaction 10 minutes；
4.3 experimental result
Prolease activity=5.2U/ml in Lactobacillus paracasei L267 metabolites.
5. applications of the Lactobacillus paracasei L267 in growing and fattening pigs feeding.
Lactobacillus paracasei L267 can be applied in growing and fattening pigs feeding.Based on the Lactobacillus paracasei L267 of the present invention,
It after being assembled by a certain percentage with other probiotics, is added in fattening pannage, the feedstuff-meat ratio of growing and fattening pigs and dead can be reduced
Rate is died, the use of antibiotic is reduced.The ratio that assembles between wherein each probiotics is：Lactobacillus paracasei：Lactobacillus acidophilus：It is dynamic
Object Bifidobacterium lactis spp：Lactobacillus casei=(2~6):(0.5~1.5):(0.5~1.5):(0.5~1).
5.1 experimental method
The growing and fattening pigs 20 for selecting weight 40kg or so, are randomly divided into control group and experimental group, every group each 10, are raised
Feed experiment.Wherein control group fed basal diet；Test group feeds ‰ lactic bacteria composition of basal diet+1.
5.2 experimental result
After test result shows the growing and fattening pigs special lactobacillus composition of addition 1 ‰：The utilization rate of feed is improved,
Test group reduces 13.18% than the feed-weight ratio of control group；The death rate reduces, and test group is reduced than the death rate of control group
87.5%；It delivers for sale in advance, test group delivers number of days for sale than control group and is advanced by 9 days.Detailed results see the table below 8：
Table 8：Growing and fattening pigs feeding experiment result
| Test group | Control group | |
| Marketing age (day) | 168 | 177 |
| The death rate (%) | 2 | 16 |
| Feedstuff-meat ratio | 2.7:1 | 3.11:1 |
After growing and fattening pigs it can be seen from upper table 8 have eaten lactic bacteria composition provided by the invention, marketing time is carried
Before, the death rate reduces.Illustrate that lactic bacteria composition is a kind of feed of health.
In conclusion the present invention is isolated from the ensilage of spontaneous fermentation, it is identified as Lactobacillus paracasei
L267.The Lactobacillus paracasei L267 has higher viable count after high density fermentation, reduces fermentation costs；To human body
Gastrointestinal environment has good tolerance, and can produce protease and help digest；Can Escherichia coli antibacterial liquid, food, drug,
It is applied in health products, food supplement；The feedstuff-meat ratio and the death rate of growing and fattening pigs can be reduced by being added in fattening pannage, be reduced anti-
The use of raw element.
The foregoing is merely the preferable specific implementation modes of the present invention, but scope of protection of the present invention is not limited thereto,
The change or replacement that any one skilled in the art is expected in the technical scope disclosed by the present invention should all be contained
Lid is within protection scope of the present invention.
SEQUENCE LISTING 1
<110>Beijing Bojinyuan Biotechnology Co., Ltd.
<120>A kind of Lactobacillus paracasei and its application
<130> 20180328
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 462
<212> DNA
<213>Lactobacillus paracasei L267 pheS gene orders
<400> 1
tattgttcgc tgacgtagca cattgggatg caccatgccg gcacctaaca cttcaatcca 60
accggtatat ttgcaaaccg ggcaaccttt accgccgcaa cgaaaacagg aaacatcaac 120
ttcaacggat ggctccgtaa atggaaaata acttggccgc aagcgaattg tccgatcttt 180
accaaaaacg tgttggcaca tggccaacaa ggttcccttt agatcagcca tggttatatg 240
cttgtcaatg acgagtcctt ccatctggtg aaactgatgg ctatgagtag catcgtcgtc 300
atcacgtcga taaaccaccc cagggctaat cattttcagc ggtcctttgg taaagtcgtg 360
cttttccatt gtccgcgcct gcattggact tgtctgcgag cgcatgagta actcattggt 420
aatgtaaaaa gtctcttgca tatccgaagc agggatgatt tt 462
SEQUENCE LISTING 2
<110>Beijing Bojinyuan Biotechnology Co., Ltd.
<120>A kind of Lactobacillus paracasei and its application
<130> 20180328
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1256
<212> DNA
<213>Lactobacillus paracasei L267 16SrRNA
<400> 1
gggcggtgtg tacaaggccc gggaacgtat tcaccgcggc gtgctgatcc gcgattacta 60
gcgattccga cttcgtgtag gcgagttgca gcctacagtc cgaactgaga atggctttaa 120
gagattagct tgacctcgcg gtctcgcaac tcgttgtacc atccattgta gcacgtgtgt 180
agcccaggtc ataaggggca tgatgatttg acgtcatccc caccttcctc cggtttgtca 240
ccggcagtct tactagagtg cccaactaaa tgctggcaac tagtcataag ggttgcgctc 300
gttgcgggac ttaacccaac atctcacgac acgagctgac gacaaccatg caccacctgt 360
cattttgccc ccgaagggga aacctgatct ctcaggtgat caaaagatgt caagacctgg 420
taaggttctt cgcgttgctt cgaattaaac cacatgctcc accgcttgtg cgggcccccg 480
tcaattcctt tgagtttcaa ccttgcggtc gtactcccca ggcggaatgc ttaatgcgtt 540
agctgcggca ctgaagggcg gaaaccctcc aacacctagc attcatcgtt tacggcatgg 600
actaccaggg tatctaatcc tgttcgctac ccatgctttc gagcctcagc gtcagttaca 660
gaccagacag ccgccttcgc cactggtgtt cttccatata tctacgcatt tcaccgctac 720
acatggagtt ccactgtcct cttctgcact caagtttccc agtttccgat gcgcttcctc 780
ggttaagccg agggctttca catcagactt aaaaaaccgc ctgcgctcgc tttacgccca 840
ataaatccgg ataacgcttg ccacctacgt attaccgcgg ctgctggcac gtagttagcc 900
gtggctttct ggttggatac cgtcacgccg acaacagtta ctctgccgac cattcttctc 960
caacaacaga gttttacgac ccgaaagcct tcttcactca cgcggcgttg ctccatcaga 1020
cttgcgtcca ttgtggaaga ttccctactg ctgcctcccg taggagtttg ggccgtgtct 1080
cagtcccaat gtggccgatc aacctctcag ttcggctacg tatcatcgcc ttggtgagcc 1140
attacctcac caactagcta atacgccgcg ggtccatcca aaagcgatag cttacgccat 1200
ctttcagcca agaaccatgc ggttcttgga tctatgcggt attagcatct gtttcc 1256
Claims (10)
1. a kind of Lactobacillus paracasei, it is characterised in that：Lactobacillus paracasei (Lactobacillus paracasei) L267,
Preserving number is：CGMCC No.14868, the deposit date is on November 08th, 2017, depositary institutions：Chinese microorganism strain management
Committee's common micro-organisms center.
2. Lactobacillus paracasei as described in claim 1, it is characterised in that：The Lactobacillus paracasei L267 to hydrochloric acid in gastric juice and
Cholate has tolerance.
3. Lactobacillus paracasei as described in claim 1, it is characterised in that：The Lactobacillus paracasei L267 lays eggs white
The ability of enzyme.
4. Lactobacillus paracasei as described in claim 1, it is characterised in that：The Lactobacillus paracasei L267 is to large intestine bar
Bacterium flora has inhibiting effect.
5. a kind of preparation method of Escherichia coli antibacterial liquid, it is characterised in that：It is placed on the as described in claim 1 of low temperature refrigerator
Lactobacillus paracasei L267 be put into 40~50 DEG C of water-baths and thaw, be aseptically inoculated into 9# fluid nutrient mediums, 37
DEG C constant temperature incubation 15h activated for 2~3 generations；The method that activated Lactobacillus paracasei L267 zymotic fluids are used into low-temperature centrifugation,
Supernatant is prepared, Escherichia coli antibacterial liquid is obtained.
6. a kind of food, which is characterized in that with the Lactobacillus paracasei system of CGMCC No.14868 described in claim 1
It is standby.
7. a kind of lactic bacteria composition, which is characterized in that comprising deposit number described in claim 1 be CGMCC No.14868
Lactobacillus paracasei and acceptable auxiliary material.
8. lactic bacteria composition as claimed in claim 7, which is characterized in that further include in the lactic bacteria composition：Acidophilus breast
Bacillus, bifidobacterium animalis subspecies and Lactobacillus casei；The additive amount of each component is：Lactobacillus paracasei：Lactobacillus acidophilus：
Bifidobacterium animalis subspecies：Lactobacillus casei=2~6：0.5~1.5：0.5~1.5：0.5~1.
9. lactic bacteria composition as claimed in claim 7, which is characterized in that auxiliary material is selected from lactose, dry starch, dextrin, ring
Dextrin, sodium carboxymethyl starch, carboxy starch propionic ester, microcrystalline cellulose, carboxymethyl cellulose, maltodextrin, magnesium stearate.
10. a kind of application of lactic bacteria composition as claimed in claim 7 in feeder for fat pig.
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