CN107488604A - MRS culture mediums, its preparation method and Isolation and purification method are used in lactic acid bacteria separation - Google Patents
MRS culture mediums, its preparation method and Isolation and purification method are used in lactic acid bacteria separation Download PDFInfo
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- CN107488604A CN107488604A CN201610408058.9A CN201610408058A CN107488604A CN 107488604 A CN107488604 A CN 107488604A CN 201610408058 A CN201610408058 A CN 201610408058A CN 107488604 A CN107488604 A CN 107488604A
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- lactic acid
- acid bacteria
- culture mediums
- mrs culture
- separation
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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
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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
- C12N1/02—Separating microorganisms from their culture media
Abstract
This application discloses a kind of separation of lactic acid bacteria to use MRS culture mediums, its preparation method and Isolation and purification method, and MRS culture mediums, using water 1000mL as relative quantity, other raw materials include：8~10g of peptone；4~5g of yeast extract；10~12g of beef extract；19~21g of glucose；3~4g of sodium acetate；1.5~1.7g of dibasic ammonium citrate；1.1~1.3mL of Tween 80；0.45~0.47g of magnesium sulfate；0.03~0.04g of manganese sulfate；2~2.2g of dipotassium hydrogen phosphate；18~22g of agar.The lactic acid bacteria screening and culturing medium of the present invention is more beneficial for distinguishing lactic acid bacteria object bacteria, strengthens the suppression to varied bacteria growing, improve the screening efficiency of lactic acid bacteria relative to the culture medium of the separating lactic acid bacterium of routine.
Description
Technical field
The application is related to a kind of lactic acid bacteria separation MRS culture mediums, its preparation method and separation and purifying side
Method.
Background technology
Lactic acid bacteria refers to the one kind of fermenting carbohydrate primary product for lactic acid without gemma, gram-positive bacterium
General name, be grown in anaerobism or amphimicrobian environment.Lactic acid bacteria is a kind of custom call, not
It is the title on microbial taxonomy.This is the quite numerous and jumbled bacterium of a group, and classification has hundreds of category, very
It is difficult using whether producing criteria for classification of the lactic acid as bacterium.Lactobacillus cell form have spherical, near-spherical,
Shaft-like or rod-short etc..Current study show that lactic acid bacteria is caused a disease except indivedual category to people, animal, it is most
To person poultry harmless, and Gram-positive lactic acid bacteria occupies the overwhelming majority, the newborn bar such as widely applied at present
Pseudomonas, Pediococcus, Leuconostoc, lactococcus, Bifidobacterium, enterococcus spp etc. are all leather
Lan Shi is positive.Substantial amounts of research shows that this kind of lactic acid bacteria can reduce pH value in animal body, ammonia of degrading,
The harmful substances such as indoles, scatol, strengthen the immunologic function of body, produce antibacterial metabolite, such as breast
Sour bacterium peptide, bacteriocin, lactic acid, hydrogen peroxide, acetic acid etc., so as to prevent and suppress the intrusion of pathogenic bacteria
And field planting, normal microecological balance in enteron aisle is maintained, improves the resistance against diseases of body, it is blue to many leather
Family name's positive bacteria (G+) and Gram-negative bacteria (G-) have strong inhibitory action, can suppress spoilage organisms in enteron aisle
Growth and breeding and spoilage product generation.
The content of the invention
It is an object of the invention to provide a kind of lactic acid bacteria separation MRS culture mediums, its preparation method and divide
From and purification process, to overcome deficiency of the prior art.
To achieve the above object, the present invention provides following technical scheme：
The embodiment of the present application discloses a kind of lactic acid bacteria separation MRS culture mediums, and ginseng is used as using water 1000mL
Examine, other raw materials include：
8~10g of peptone；
4~5g of yeast extract；
10~12g of beef extract；
19~21g of glucose；
3~4g of sodium acetate；
1.5~1.7g of dibasic ammonium citrate；
1.1~1.3mL of Tween 80；
0.45~0.47g of magnesium sulfate；
0.03~0.04g of manganese sulfate；
2~2.2g of dipotassium hydrogen phosphate；
18~22g of agar.
Preferably, in MRS culture mediums are used in above-mentioned lactic acid bacteria separation, in the MRS culture mediums also
Including 2% calcium carbonate.
Preferably, in MRS culture mediums are used in above-mentioned lactic acid bacteria separation, the pH of the MRS culture mediums
It is adjusted to 6.2~6.4.
Preferably, in MRS culture mediums are used in above-mentioned lactic acid bacteria separation, MRS is trained at 121 DEG C
Base is supported to sterilize 15~20 minutes.
Accordingly, disclosed herein as well is a kind of preparation method of lactic acid bacteria separation MRS culture mediums, bag
Include step：
(1) each component is weighed according to formula, by peptone, yeast extract, beef extract, glucose, acetic acid
Sodium, dibasic ammonium citrate, Tween 80, magnesium sulfate, manganese sulfate, dipotassium hydrogen phosphate, agar are put into container,
Acid solution is added, adjusts pH to 6.2~6.4,121 DEG C sterilize 15~20 minutes；
It is (2) polymyxins, nalidixic acid, ascorbic acid, Cys-HCl is soluble in water,
Filtration sterilization；
(3) mixture after step (1) is sterilized is cooled to 30~50 DEG C, what same step (2) obtained
Solution and the compound acid-base indicator prepared in advance mix mixing.
Disclosed herein as well is a kind of separation of culture medium and the method for purified lactic acid bacterium, the separation of lactic acid bacteria is adopted
With plate dilution method, the bacterial strain obtained to separation uses plate streak.
Compared with prior art, the advantage of the invention is that：The lactic acid bacteria screening and culturing medium of the present invention is relative
In the culture medium of the separating lactic acid bacterium of routine, it is more beneficial for distinguishing lactic acid bacteria object bacteria, strengthens to miscellaneous bacteria
The suppression of growth, improve the screening efficiency of lactic acid bacteria.
Embodiment
The present invention is described further by the following example：, can be more preferably geographical according to following embodiments
The solution present invention.However, as it will be easily appreciated by one skilled in the art that specific material described by embodiment
The present invention is merely to illustrate than, process conditions and its result, without should be also without limitation on claims
In the described in detail present invention.
Embodiment 1
MRS culture mediums are used in lactic acid bacteria separation：Peptone 10g, yeast extract 5g, beef extract 10g, grape
Sugared 20g, sodium acetate 4g, dibasic ammonium citrate 1.5g, Tween 80 1mL, magnesium sulfate 0.5g, manganese sulfate 0.04g,
Dipotassium hydrogen phosphate 2g, agar 20g, water 1000mL.
Also include 2% calcium carbonate in MRS culture mediums.
The pH of MRS culture mediums is adjusted to 6.2~6.4.
To MRS medium sterilizations 15~20 minutes at 121 DEG C.
Culture medium is prepared as follows：
(1) each component is weighed according to formula, by peptone, yeast extract, beef extract, glucose, acetic acid
Sodium, dibasic ammonium citrate, Tween 80, magnesium sulfate, manganese sulfate, dipotassium hydrogen phosphate, agar are put into container,
Acid solution is added, adjusts pH to 6.2~6.4,121 DEG C sterilize 15~20 minutes；
It is (2) polymyxins, nalidixic acid, ascorbic acid, Cys-HCl is soluble in water,
Filtration sterilization；
(3) mixture after step (1) is sterilized is cooled to 50 DEG C, the solution that same step (2) obtains
And the compound acid-base indicator prepared in advance mixes mixing.
Utilize above-mentioned culture medium Isolation and purification method：
The separation of lactic acid bacteria uses plate dilution method, and the bacterial strain obtained to separation uses plate streak.
Embodiment 2
MRS culture mediums are used in lactic acid bacteria separation：Peptone 9g, yeast extract 5g, beef extract 10g, glucose
21g, sodium acetate 4g, dibasic ammonium citrate 1.5g, Tween 80 1.3mL, magnesium sulfate 0.5g, manganese sulfate 0.04g,
Dipotassium hydrogen phosphate 2.2g, agar 20g, water 1000mL.
Also include 2% calcium carbonate in MRS culture mediums.
The pH of MRS culture mediums is adjusted to 6.2~6.4.
To MRS medium sterilizations 15~20 minutes at 121 DEG C.
Culture medium is prepared as follows：
(1) each component is weighed according to formula, by peptone, yeast extract, beef extract, glucose, acetic acid
Sodium, dibasic ammonium citrate, Tween 80, magnesium sulfate, manganese sulfate, dipotassium hydrogen phosphate, agar are put into container,
Acid solution is added, adjusts pH to 6.2~6.4,121 DEG C sterilize 15~20 minutes；
It is (2) polymyxins, nalidixic acid, ascorbic acid, Cys-HCl is soluble in water,
Filtration sterilization；
(3) mixture after step (1) is sterilized is cooled to 50 DEG C, the solution that same step (2) obtains
And the compound acid-base indicator prepared in advance mixes mixing.
Utilize above-mentioned culture medium Isolation and purification method：
The separation of lactic acid bacteria uses plate dilution method, and the bacterial strain obtained to separation uses plate streak.
Embodiment 3
MRS culture mediums are used in lactic acid bacteria separation：Peptone 10g, yeast extract 4g, beef extract 11g, grape
Sugared 20g, sodium acetate 4g, dibasic ammonium citrate 1.7g, Tween 80 1mL, magnesium sulfate 0.45g, manganese sulfate
0.03g, dipotassium hydrogen phosphate 2g, agar 22g, water 1000mL.
Also include 2% calcium carbonate in MRS culture mediums.
The pH of MRS culture mediums is adjusted to 6.2~6.4.
To MRS medium sterilizations 15~20 minutes at 121 DEG C.
Culture medium is prepared as follows：
(1) each component is weighed according to formula, by peptone, yeast extract, beef extract, glucose, acetic acid
Sodium, dibasic ammonium citrate, Tween 80, magnesium sulfate, manganese sulfate, dipotassium hydrogen phosphate, agar are put into container,
Acid solution is added, adjusts pH to 6.2~6.4,121 DEG C sterilize 15~20 minutes；
It is (2) polymyxins, nalidixic acid, ascorbic acid, Cys-HCl is soluble in water,
Filtration sterilization；
(3) mixture after step (1) is sterilized is cooled to 50 DEG C, the solution that same step (2) obtains
And the compound acid-base indicator prepared in advance mixes mixing.
Utilize above-mentioned culture medium Isolation and purification method：
The separation of lactic acid bacteria uses plate dilution method, and the bacterial strain obtained to separation uses plate streak.
Finally, it is to be noted that, term " comprising ", "comprising" or its any other variant are intended to
Nonexcludability includes, so that process, method, article or equipment including a series of elements are not
Only include those key elements, but also the other element including being not expressly set out, or it is this also to include
Kind of process, method, article or the intrinsic key element of equipment.
Claims (6)
1. MRS culture mediums are used in a kind of lactic acid bacteria separation, it is characterised in that are used as phase using water 1000mL
To amount, other raw materials include：
8~10g of peptone；
4~5g of yeast extract；
10~12g of beef extract；
19~21g of glucose；
3~4g of sodium acetate；
1.5~1.7g of dibasic ammonium citrate；
1.1~1.3mL of Tween 80；
0.45~0.47g of magnesium sulfate；
0.03~0.04g of manganese sulfate；
2~2.2g of dipotassium hydrogen phosphate；
18~22g of agar.
2. MRS culture mediums are used in lactic acid bacteria separation according to claim 1, it is characterised in that：Institute
State also include in MRS culture mediums 2% calcium carbonate.
3. MRS culture mediums are used in lactic acid bacteria separation according to claim 1, it is characterised in that：Institute
The pH for stating MRS culture mediums is adjusted to 6.2~6.4.
4. MRS culture mediums are used in lactic acid bacteria separation according to claim 1, it is characterised in that：
To MRS medium sterilizations 15~20 minutes at 121 DEG C.
5. the preparation method of the lactic acid bacteria separation MRS culture mediums described in claim 1, its feature exist
In, including step：
(1) each component is weighed according to formula, by peptone, yeast extract, beef extract, glucose, acetic acid
Sodium, dibasic ammonium citrate, Tween 80, magnesium sulfate, manganese sulfate, dipotassium hydrogen phosphate, agar are put into container,
Acid solution is added, adjusts pH to 6.2~6.4,121 DEG C sterilize 15~20 minutes；
It is (2) polymyxins, nalidixic acid, ascorbic acid, Cys-HCl is soluble in water,
Filtration sterilization；
(3) mixture after step (1) is sterilized is cooled to 30~50 DEG C, what same step (2) obtained
Solution and the compound acid-base indicator prepared in advance mix mixing.
6. the method with purified lactic acid bacterium is separated using any described culture medium of Claims 1-4 4, its
It is characterised by：The separation of lactic acid bacteria uses plate dilution method, and the bacterial strain obtained to separation uses plate streaking
Method.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337836A (en) * | 2018-10-26 | 2019-02-15 | 贵州茅台酒股份有限公司 | A kind of lactic acid bacteria isolation medium and preparation method thereof and lactic acid bacterial screening method |
| CN109371100A (en) * | 2018-11-20 | 2019-02-22 | 四川农业大学 | A kind of culture medium and its method for the detection of vinegar aerogenic bacteria |
| CN109593676A (en) * | 2018-12-21 | 2019-04-09 | 江苏大学 | A kind of culture medium and preparation method thereof separated for microorganism in sauerkraut fermentation liquid |
| CN111423997A (en) * | 2020-03-31 | 2020-07-17 | 广州市妇女儿童医疗中心（广州市妇幼保健院、广州市儿童医院、广州市妇婴医院、广州市妇幼保健计划生育服务中心） | Group B streptococcus enriched broth and application thereof |
| CN111979127A (en) * | 2020-09-03 | 2020-11-24 | 青海雪峰牦牛乳业有限责任公司 | Method for separating lactic acid bacteria from yak triton |
| CN114009767A (en) * | 2021-11-17 | 2022-02-08 | 百瑞源枸杞股份有限公司 | Fresh medlar primary pulp fermentation product and preparation method and application thereof |
-
2016
- 2016-06-12 CN CN201610408058.9A patent/CN107488604A/en not_active Withdrawn
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337836A (en) * | 2018-10-26 | 2019-02-15 | 贵州茅台酒股份有限公司 | A kind of lactic acid bacteria isolation medium and preparation method thereof and lactic acid bacterial screening method |
| CN109337836B (en) * | 2018-10-26 | 2021-11-26 | 贵州茅台酒股份有限公司 | Lactobacillus isolation medium, preparation method thereof and lactobacillus screening method |
| CN109371100A (en) * | 2018-11-20 | 2019-02-22 | 四川农业大学 | A kind of culture medium and its method for the detection of vinegar aerogenic bacteria |
| CN109371100B (en) * | 2018-11-20 | 2023-09-22 | 四川农业大学 | Culture medium for detecting vinegar gas-producing bacteria and method thereof |
| CN109593676A (en) * | 2018-12-21 | 2019-04-09 | 江苏大学 | A kind of culture medium and preparation method thereof separated for microorganism in sauerkraut fermentation liquid |
| CN109593676B (en) * | 2018-12-21 | 2023-04-07 | 江苏大学 | Culture medium for separating microorganisms from pickled Chinese cabbage fermentation liquor and preparation method thereof |
| CN111423997A (en) * | 2020-03-31 | 2020-07-17 | 广州市妇女儿童医疗中心（广州市妇幼保健院、广州市儿童医院、广州市妇婴医院、广州市妇幼保健计划生育服务中心） | Group B streptococcus enriched broth and application thereof |
| CN111423997B (en) * | 2020-03-31 | 2022-02-11 | 广州市妇女儿童医疗中心（广州市妇幼保健院、广州市儿童医院、广州市妇婴医院、广州市妇幼保健计划生育服务中心） | Group B streptococcus enriched broth and application thereof |
| CN111979127A (en) * | 2020-09-03 | 2020-11-24 | 青海雪峰牦牛乳业有限责任公司 | Method for separating lactic acid bacteria from yak triton |
| CN114009767A (en) * | 2021-11-17 | 2022-02-08 | 百瑞源枸杞股份有限公司 | Fresh medlar primary pulp fermentation product and preparation method and application thereof |
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Application publication date: 20171219 |