CN106819107A - Leben with the effect of whole intestines and preparation method thereof - Google Patents
Leben with the effect of whole intestines and preparation method thereof Download PDFInfo
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- CN106819107A CN106819107A CN201611248415.6A CN201611248415A CN106819107A CN 106819107 A CN106819107 A CN 106819107A CN 201611248415 A CN201611248415 A CN 201611248415A CN 106819107 A CN106819107 A CN 106819107A
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- leben
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- lactobacillus plantarum
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
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Abstract
The present invention relates to a kind of leben with the effect of whole intestines, milk is equipped with galactooligosaccharide, it is obtained after being fermented through Lactobacillus plantarum JMCC0017, Lactobacillus plantarum JMCC0017 is the separation screening from Xinjiang of China traditional zymotic dairy product, the bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number on March 31st, 2016：CGMCC NO.12323, galactooligosaccharide and Lactobacillus plantarum JMCC0017 coordinateds, promote the cultivation effect of Lactobacillus plantarum JMCC0017, improve whole intestines effect, Bifidobacterium is also included in inventive formulation, using Lactobacillus plantarum JMCC0017 and the mutualism relation of Bifidobacterium, the whole gut function of acidified milk is further improved.
Description
Technical field
The invention belongs to dairy products field, and in particular to a kind of leben with the effect of whole intestines；It is simultaneously of the invention
Further relate to a kind of preparation method of the leben should with the effect of whole intestines.
Background technology
Enteron aisle not only has digestion and absorption function, also barrier function.Complete gut barrier includes 3 parts：Machinery
Barrier, biological barrier, immunization barrier.Mechanical barrier refers to complete Intestinal epithelial cells structure tightly attached to each other, raw
Thing barrier includes biochemical substances barrier and microbial barrier, and microbial barrier therein is main by colonizing in human body intestinal canal about
More than 500 kinds of enteric microorganism is constituted, and these symbiosis floras adhere to by with intestinal mucosa, with reference to, the formation one that interacts such as chimeric
Individual complete mycoderm, constitutes the biological barrier of enteron aisle.Majority is lactic acid bacteria in the middle of these micropopulations, and they pass through surface
Together with some specific compositions stick with the gastrointestinal mucosal of human body, repel pathogenic bacteria sticking and invade to normal cell
Attack.Lactic acid bacteria plays an important roll to the adhesion of body cell for regulating intestinal canal micro-ecological environment.
The content of the invention
It is an object of the invention to provide a kind of leben with the effect of whole intestines.
A kind of leben with the effect of whole intestines, is prepared from the following raw materials in parts by weight：
Raw milk 60-100 parts, 0.2-15 parts, galactooligosaccharide 2-30 parts of Lactobacillus plantarum JMCC0017.
Further, the Lactobacillus plantarum JMCC0017 is 5-10 parts.
Further, described galactooligosaccharide 10-20 parts.
Further, the leben also includes following parts by weight raw material：Bifidobacterium 0.2-15 parts.
Further, the Lactobacillus plantarum JMCC0017 and the inoculative proportion of Bifidobacterium are 1:1.
Leben with the effect of whole intestines of the present invention has following beneficial effect：
1st, Lactobacillus plantarum JMCC0017 is the separation screening from Xinjiang of China traditional zymotic dairy product, and the bacterial strain is
China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number are preserved on March 31st, 2016：
CGMCC NO.12323, depositary institution address is：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.Lactobacillus plantarum JMCC0017
Be one plant with tolerable temperature change, high activity, it is weak after acid, without special plant local flavor, good acid and bile salt tolerance and anti-oxidant
The strain excellent of performance, the bacterial strain also has preferable enteron aisle definite value ability.
2nd, Lactobacillus plantarum JMCC0017 can promote enterocyte to secrete tight junction protein occludin, regulating intestinal canal barrier
Permeability, maintain the 26S Proteasome Structure and Function of enteric epithelium, prevent the toxic material such as pathogen from entering internal.
3rd, galactooligosaccharide and Lactobacillus plantarum JMCC0017 coordinateds, promote the increasing of Lactobacillus plantarum JMCC0017
Grow, so as to improve the whole intestines effect of leben.
4th, further, mutualism between Lactobacillus plantarum JMCC0017 and Bifidobacterium, interaction promotes growth, activity
The propionic acid bacteria growing that the lactic acid that Lactobacillus plantarum JMCC0017 is produced can promote in enteron aisle, and one of product of bacterium acidi propionici propionic acid
Salt is a kind of bifidus factor, and then promotes the propagation of the Bifidobacterium in enteron aisle, and then strengthens whole gut function.
The preparation method of the leben with the effect of whole intestines, comprises the following steps：
A, with material milk：Galactooligosaccharide is added in raw milk, stirring to fully dissolving must match somebody with somebody material milk after homogeneous, sterilization；
B, fermentation：41 ± 1.0 DEG C will be cooled to material milk, add Lactobacillus plantarum JMCC0017 strains, stirring 30min will make
Bacterium powder is fully dissolved, 41 ± 1.0 DEG C of fermentations, and fermentation 4-5h starts to survey acid, and fermentation, stirring demulsification, hair are stopped when acidity is 80 ° of T
Kefir milk is cooled to 10-30 DEG C, filling, stored refrigerated.
Further, in step a homogeneous, process for sterilizing are as follows：
Homogeneous：58-70 DEG C of homogenizing temperature, homogenization pressure 16-20MPa；
It is sterilized：136 ± 2 DEG C of sterilization temperature, sterilizing time 4-15s；
Further, the refrigerated storage temperature in step a is 2 DEG C -6 DEG C.
Using preparation method of the invention, can further ensure that there are the leben being prepared into preferably whole intestines to make
With.
Specific embodiment
Embodiments of the present invention are illustrated below by way of specific instantiation, but and is not used to limit reality of the invention
Apply scope.
Embodiment 1
Lactobacillus plantarum (Lactobacillus plantarum) JMCC0017 and its screening technique
A kind of Lactobacillus plantarum (Lactobacillus plantarum) JMCC0017, it is sent out from Xinjiang of China tradition
The probiotics filtered out in kefir milk product, the bacteria strain has been preserved in the common micro- life of China Committee for Culture Collection of Microorganisms
Thing center, deposit number：CGMCC NO.12323.
The screening technique of Lactobacillus plantarum (Lactobacillus plantarum) JMCC0017, according to the following steps sequentially
Carry out：
A, sample collection
The traditional fermented dairy product in 25mL Xinjiang is taken, is added in 225mL SPSSs, fully mixed, mixed
Liquid sample is closed,
B, strain isolation, purifying
1mL mixed liquor samples are taken, gradient dilution is pressed with SPSS, 10 are diluted to respectively-3、10-4、10-5Times, obtain
To the sample diluting liquid of various concentrations；Draw 0.1mL sample diluting liquids to coat on solid MRS culture medium flat plates, 37 DEG C of anaerobism
After culture 72h, colony characteristicses and reference pertinent literature picture according to standard plant lactobacillus, the alternative bacterium colony of picking continue to train
Support, continuous passage is no less than three times, obtain the bacterium colony of purifying.
The identification of c, bacterial strain
From separate bacterial strain, selection Gram-positive, H2O2Negative, sour, anaerogenic bacterial strain can be produced,
The bacterium colony of purifying is placed in 20% aseptic glycerine in -70 DEG C of preservations, while be inoculated with MRS agar mediums test tube slant being used for
It is interim to preserve, above-mentioned bacterial strains are expanded by DNA extractions, PCR, 16SrRNA gene orders, the experimental data such as pheS gene orders
Comprehensive analysis, qualification result is Lactobacillus plantarum JMCC0017.
The 16SrRNA gene orders of Lactobacillus plantarum JMCC0017 are SEQ ID NO:1.
The pheS gene orders of Lactobacillus plantarum JMCC0017 are SEQ ID NO:2.
The screening technique of Lactobacillus plantarum JMCC0017, MRS agar mediums are consisted of the following composition in step c：
Casein peptone：10g；Beef extract：10g；Yeast extract：5g；Glucose：20g；Sodium acetate：5g；Lemon acid diamine：2g；
Tween-80：1g；K2HPO4：2g；MgSO4·7H2O：0.2g；MnSO4·7H2O：0.05g；Agar：15g；Distilled water：
1000mL；The culture medium adjusts pH6.8 ± 0.1.
The biological characteristics of Lactobacillus plantarum JMCC0017 is as follows：
1st, cellular morphology and physical and chemical experiment result
The cellular morphology and physical and chemical experiment result of Lactobacillus plantarum JMCC0017 are as shown in table 1 below：
The cellular morphology and physical and chemical experiment result of the JMCC0017 of table 1
As shown in Table 1, Lactobacillus plantarum JMCC0017 be Gram-positive, it is shaft-like, without gemma, catalase and oxidation
The negative bacterial strain of enzyme experiment；Bacterial strain glucose aerogenesis experiment is negative, and the experiment of sodium gluconate aerogenesis is positive；Carbohydrate is produced
In sour characteristic, "+" represents that fermentation is utilized, and "-" represents that azymic is utilized.
2nd, antibiotics sensitivity experimental result
The antibiotics sensitivity experimental result of table 2
| Antibiotic title | Sensitiveness | Antibiotic title | Sensitiveness | Antibiotic title | Sensitiveness |
| Piperacillin/Tazobactam Sodium | S | SM 7338 | S | Rifampin | S |
| Spectinomycin | R | Vancomycin | R | Tetracycline | M |
| Ciprofloxacin | R | Furantoin | S | SMZco | S |
| Penicillin | S | The Qusong of cephalo three | S | Amikacin | R |
| Erythromycin | S | Cefaclor | S | Cefotaxime | S |
| Chloramphenicol | S | Cephazoline | S | Cefoxitin | S |
| Azithromycin | M | CTX | S | Cefoperazone | S |
| Clindamycin | M | The western quinoline of ammonia benzyl | S | Gentamicin | R |
| Doxycycline | S | Head born of the same parents' furan is new | S | OXA | S |
| CLA | S | Minocycline | S |
Lactobacillus plantarum JMCC0017 is determined to 29 kinds of sensitiveness of antibiotic, is as a result shown, the strain is to test
21 kinds of antibiotic performances are sensitive, insensitive to 5 kinds of antibiotic.
Embodiment 2-8
Embodiment 2-8 shows the material content of the leben with the effect of whole intestines, as shown in table 3：
Each embodiment content of raw material table of table 3
Comparative example 1-3
Table 4 show with whole intestines effect leben material content three comparative examples, its material composition and
Proportioning is as follows：
The comparative example content of raw material table of table 4
The leben with the effect of whole intestines of example is performed as described above, its preparation method is comprised the following steps：
A, with material milk：Galactooligosaccharide, stirring to fully dissolving, 58-70 DEG C of homogeneous, homogenization pressure are added in raw milk
16-20MPa, 136 ± 2 DEG C of sterilization 4-15s, obtains with material milk；
B, fermentation：41 ± 1.0 DEG C will be cooled to material milk, add Lactobacillus plantarum JMCC0017 strains, stirring 30min will make
Bacterium powder is fully dissolved, 41 ± 1.0 DEG C of fermentations, and fermentation 4-5h starts to survey acid, and fermentation, stirring demulsification, hair are stopped when acidity is 80 ° of T
Kefir milk is cooled to 10-30 DEG C, it is filling, 2 DEG C -6 DEG C it is stored refrigerated.
Leben antiflatulent prepared by Lactobacillus plantarum JMCC0017 and each embodiment, comparative example can be tested：
1st, Lactobacillus plantarum JMCC0017 intestinal colonisations aptitude tests.
Lactobacillus plantarum JMCC0017 has preferable intestinal colonisation ability, and evaluation model, analysis are adhered to by enterocyte
The adhesive capacity of 10 strains of lactic acid bacteria.Enterocyte Caco-2 is laid in Tissue Culture Plate in advance, the lactic acid bacteria that will have been activated
With PBS 2 times after thalline centrifugation, with the DMEM culture mediums of antibiotic-free it is resuspended after, be laid on cell upper strata and co-culture 2h.Wash away
1mL 1% (v/v) Triton X-100Stock Solution are added after uncombined lactic acid bacteria, makes the same cell of thalline of adhesion
Separate, be placed in after gradient dilution on MRS flat boards after cultivating 48h and counted.Total bacteria count/every hole inner cell in adhesion results=every hole
Number, the lactic acid bacteria number adhered on each cell is more, shows that its colonization ability is stronger, as a result as shown in table 5 below, plant breast bar
Bacterium JMCC0017 can be adhered 31 bacterium on each enterocyte surface, and adhesive capacity is very strong.
The intestinal colonisation aptitude tests data of table 5
| Strain number | Bacterium number/hole | Bacterium number/cell |
| JMCC0017 | 31.05 | |
| N1207 | 4.02 | |
| N1115 | 6.00 | |
| DL115107 | 5.08 | |
| N3114 | 34.18 | |
| DL112109 | 4.92 | |
| N3207 | 29.49 | |
| Rb111116 | 3.32 | |
| DL112109 | 6.15 | |
| KL24 | 7.13 |
2nd, each embodiment and the whole intestines aptitude tests of comparative example acidified milk.
Acidified milk gavage experiment mice prepared by above-described embodiment and comparative example, selection mouse 20, male and female half and half, often
Group 20；Difference each embodiment of gavage and comparative example acidified milk, gavage amount are 8g/kg BW, once a day, continuous 14 days, gavage
Microflora in front and rear inspection mouse intestinal, it is as a result as shown in table 6 below：
Influence of the acidified milk of table 6 to the common Microflora of mouse intestinal
Understand that the acidified milk containing Lactobacillus plantarum JMCC0017 can promote bifidobacteria and lactobacillus quantity from upper table 6
Increase, both strains are intestinal beneficial bacteriums, improve function of intestinal canal, galactooligosaccharide is cooperateed with Lactobacillus plantarum JMCC0017 matches somebody with somebody
Close, promote the cultivation effect of Lactobacillus plantarum JMCC0017, improve whole intestines effect.Lactobacillus plantarum JMCC0017 and Bifidobacterium
Between mutualism, strengthen whole gut function.
SEQUENCE LISTING
<110>Shijiazhuang Jun Lebao emulsions Co., Ltd
<120>Leben with the effect of whole intestines and preparation method thereof
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 1467
<212> DNA
<213>The 16SrRNA gene orders of Lactobacillus plantarum Lactobacillus plantarum JMCC0017
<400> 1
atgcagtcga acgaactctg gtattgattg gtgcttgcat catgatttac atttgagtga 60
gtggcgaact ggtgagtaac acgtgggaaa cctgcccaga agcgggggat aacacctgga 120
aacagatgct aataccgcat aacaacttgg accgcatggt ccgagcttga aagatggctt 180
cggctatcac ttttggatgg tcccgcggcg tattagctag atggtggggt aacggctcac 240
catggcaatg atacgtagcc gacctgagag ggcaatcggc cacattggga ctgagacacg 300
gcccaaactc ctacgggagg cagcagtagg gaatcttcca caatggacga aagtctgatg 360
gagcaacgcc gcgtgagtga agaagggttt cggctcgtaa aactctgttg ttaaagaaga 420
acatatctga gagtaactgt tcaggtattg acggtattta accagaaagc cacggctaac 480
tacgtgccag cagccgcggt aatacgtagg tggcaagcgt tgtccggatt tattgggcgt 540
aaagcgagcg caggcggttt tttaagtctg atgtgaaagc cttcggctca accgaagaag 600
tgcatcggaa actgggaaac ttgagtgcag aagaggacag tggaactcca tgtgtagcgg 660
tgaaatgcgt agatatatgg aagaacacca gtggcgaagg cggctgtctg gtctgtaact 720
gacgctgagg ctcgaaagta tgggtagcaa acaggattag ataccctggt agtccatacc 780
gtaaacgatg aatgctaagt gttggagggt ttccgccctt cagtgctgca gctaacgcat 840
taagcattcc gcctggggag tacggccgca aggctgaaac tcaaaggaat tgacgggggc 900
ccgcacaagc ggtggagcat gtggtttaat tcgaagctac gcgaagaacc ttaccaggtc 960
ttgacatact atgcaaatct aagagattag acgttccctt cggggacatg gatacaggtg 1020
gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080
acccttatta tcagttgcca gcattaagtt gggcactctg gtgagactgc cggtgacaaa 1140
ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg 1200
tgctacaatg gatggtacaa cgagttgcga actcgcgaga gtaagctaat ctcttaaagc 1260
cattctcagt tcggattgta ggctgcaact cgcctacatg aagtcggaat cgctagtaat 1320
cgcggatcag catgccgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcacac 1380
catgagagtt tgtaacaccc aaagtcggtg gggtaacctt ttaggaacca gccgcctaag 1440
gtgggacaga tgattagggt gaagtcg 1467
<210> 2
<211> 450
<212> DNA
<213>The pheS gene orders of Lactobacillus plantarum Lactobacillus plantarum JMCC0017
<400> 2
caaagacgtg ctactacgca cgcagacgtc tgctgatcag ccgcggtcac ttgaaaatca 60
cgatttttct aaaggaccgc tgaaggtctt gtcacctggc cgcgtttatc ggcgtgatac 120
ggatgatgca acccattccc atcaatttca tcaaattgaa gggttagtcg tggacaagca 180
tattacgatg gctgatttga agggcacctt aattctggtt gccaagactt tgtttggcga 240
tcaattcgat gttcggctac ggccaagctt ctttccattc acggaaccat ccgtagaagc 300
tgatgtaact tgctttaatt gcaatggcaa gggctgtgca atctgtaagc aaacgggttg 360
gatcgaagta ctgggtgccg gcatggttca cccccacgtg ttagaaatgt ctggcattga 420
tccagaagaa tatggtggct ttgcctttgg 450
Claims (8)
1. it is a kind of with whole intestines effect leben, it is characterised in that the leben is by following parts by weight
Raw material is made：
Raw milk 60-100 parts, 0.2-15 parts, galactooligosaccharide 2-30 parts of Lactobacillus plantarum JMCC0017.
2. it is according to claim 1 with whole intestines effect leben, it is characterised in that the Lactobacillus plantarum
JMCC0017 is 5-10 parts.
3. it is according to claim 1 with whole intestines effect leben, it is characterised in that the galactooligosaccharide
10-20 parts.
4. it is according to claim 1 with whole intestines effect leben, it is characterised in that the leben is also
Including following parts by weight raw material：Bifidobacterium 0.2-15 parts.
5. it is according to claim 4 with whole intestines effect leben, it is characterised in that the Lactobacillus plantarum
The ratio of JMCC0017 and Bifidobacterium is 1:1.
6. the preparation method of a kind of leben with the effect of whole intestines as any one of claim 1-5, it is special
Levy and be, the method is comprised the following steps：
A, with material milk：Galactooligosaccharide is added in raw milk, stirring to fully dissolving must match somebody with somebody material milk after homogeneous, sterilization；
B, fermentation：41 ± 1.0 DEG C will be cooled to material milk, add Lactobacillus plantarum JMCC0017 strains, stirring 30min will make bacterium powder
Fully dissolving, 41 ± 1.0 DEG C of fermentations, fermentation 4-5h starts to survey acid, and fermentation, stirring demulsification, acidified milk are stopped when acidity is 80 ° of T
It is cooled to 10-30 DEG C, it is filling, stored refrigerated.
7. it is according to claim 6 with whole intestines effect leben preparation method, it is characterised in that step a
In homogeneous, process for sterilizing be：
Homogeneous：58-70 DEG C of homogenizing temperature, homogenization pressure 16-20MPa；
It is sterilized：136 ± 2 DEG C of sterilization temperature, sterilizing time 4-15s.
8. it is according to claim 6 with whole intestines effect leben preparation method, it is characterised in that step a
In refrigerated storage temperature be 2 DEG C -6 DEG C.
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ID=59115114
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107981360A (en) * | 2017-11-24 | 2018-05-04 | 华东理工大学 | A kind of application of lactobacillus plantarum X7021 |
| CN111011866A (en) * | 2019-12-12 | 2020-04-17 | 上海理工大学 | Application of prebiotics in promoting growth of lactobacillus plantarum in intestinal environment |
| CN112535214A (en) * | 2020-12-03 | 2021-03-23 | 石家庄君乐宝乳业有限公司 | Probiotic milk tablet with intestine regulating effect and preparation method thereof |
| CN112544721A (en) * | 2020-12-08 | 2021-03-26 | 石家庄君乐宝乳业有限公司 | Milk tablet for promoting growth of vital essence and preparation method thereof |
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| CN107981360A (en) * | 2017-11-24 | 2018-05-04 | 华东理工大学 | A kind of application of lactobacillus plantarum X7021 |
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| CN112535214A (en) * | 2020-12-03 | 2021-03-23 | 石家庄君乐宝乳业有限公司 | Probiotic milk tablet with intestine regulating effect and preparation method thereof |
| CN112544721A (en) * | 2020-12-08 | 2021-03-26 | 石家庄君乐宝乳业有限公司 | Milk tablet for promoting growth of vital essence and preparation method thereof |
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