CN103194417A - Lactic acid bacterium culture medium - Google Patents
Lactic acid bacterium culture medium Download PDFInfo
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- CN103194417A CN103194417A CN2013101479292A CN201310147929A CN103194417A CN 103194417 A CN103194417 A CN 103194417A CN 2013101479292 A CN2013101479292 A CN 2013101479292A CN 201310147929 A CN201310147929 A CN 201310147929A CN 103194417 A CN103194417 A CN 103194417A
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Abstract
The invention discloses a lactic acid bacterium culture medium which comprises the following components: 1) 10-12g/L of peptone, 2) 4-6g/L of yeast extract, 3) 6-22g/L of glucose, 4) 2-7g/L of acid-base buffering agent, 5) 0.15-0.4g/L of growth factor, 6) 0-10g/L of beef extract, 7) 0-15g/L of soluble starch, 0-3g/L of fructose and 9) 0-20g/L of agar, wherein the pH value of the culture medium is 5.8 to 6.9. By utilizing the culture medium, the lactic acid bacterium can grow well, and moreover the problem that nearly all of the conventional commercial MRS culture media have the inbibitional effect to helicobacter pylori is solved, so that the culture medium can be used for screening lactic acid bacteria having the inbibitional effect to the helicobacter pylori, and a real and reliable screening result can be obtained in the screening experiment; and meanwhile by utilizing the culture medium, the cost of the lactic acid bacterium culture medium is greatly lowered.
Description
Technical field
The present invention relates to a kind of substratum, particularly a kind of milk-acid bacteria substratum.
Background technology
Helicobacter pylori (Helicobacter pylori) is acknowledged as the main virulence factor of gastritis, peptide ulceration and cancer of the stomach, the common method of medically treating helicobacter pylori infection is to utilize proton pump inhibitor to add two kinds of antibiotic triple therapies, but owing to increasing of the Resistant strain that produces thereupon, make curative effect descend.Result of study shows that some specific milk-acid bacteria has inhibition and even killing action to helicobacter pylori, can alleviate even cure the microbial relative disease of helicobacter pylorus, therefore, good using value is being arranged medically, becomes international research focus gradually.
At present at home and abroad, more existing milk-acid bacteria suppresses in the body of helicobacter pylori, the experiment in vitro report about utilizing, but nearly all report is not all mentioned a fact, that be exactly in the past to the experiment of the in-vitro screening of milk-acid bacteria in employed milk-acid bacteria substratum (be widely used as the selective medium of MRS substratum as milk-acid bacteria, commercially produced product is the most common with Merck company and Oxoid company) itself to the restraining effect of helicobacter pylori.The present patent application people finds, in screening helicobacter pylori is had in the experiment in vitro of inhibiting milk-acid bacteria, as milk-acid bacteria the MRS substratum self of normal employing helicobacter pylori is had very strong restraining effect, this inhibition can cause the false appearance of screening experiment, and screening process is produced certain interference.Therefore, this area need have a kind of substratum can guarantee the good growth of milk-acid bacteria especially, can eliminate substratum itself again to the restraining effect of helicobacter pylori, thereby obtain real in-vitro screening result.
Summary of the invention
Technical problem to be solved by this invention is in order to overcome in the past when screening suppresses the milk-acid bacteria of helicobacter pylori, there is restraining effect in used milk-acid bacteria substratum itself to helicobacter pylori, thereby disturb the defective of The selection result, and provide a kind of good growth that can guarantee milk-acid bacteria, and helicobacter pylori there is not inhibiting substratum.
For solving the problems of the technologies described above, one of technical scheme provided by the invention is: a kind of milk-acid bacteria substratum, and described milk-acid bacteria substratum is made up of following component:
1) peptone 10～12g/L;
2) yeast extract 4～6g/L;
3) glucose 6～22g/L;
4) acid-base buffer agent 2～7g/L, described acid-base buffer agent are ammonium citrate and/or dipotassium hydrogen phosphate;
5) somatomedin 0.15～0.4g/L, described somatomedin is sal epsom and/or manganous sulfate;
6) beef extract 0～10g/L;
7) Zulkovsky starch 0～15g/L;
8) fructose 0～3g/L; With
9) agar 0～20g/L;
The pH value of described substratum is 5.8-6.9.
Among the present invention, contain beef extract 0～10g/L in the described milk-acid bacteria substratum; Preferably, contain beef extract 8～10g/L in the described milk-acid bacteria substratum.
Among the present invention, contain Zulkovsky starch 0～15g/L in the described milk-acid bacteria substratum; Preferably, contain Zulkovsky starch 5～15g/L in the described milk-acid bacteria substratum.
Among the present invention, contain fructose 0～3g/L in the described milk-acid bacteria substratum; Preferably, contain fructose 3g/L in the described milk-acid bacteria substratum.
Among the present invention, contain agar 0～20g/L in the described milk-acid bacteria substratum; Preferably, contain agar 10～20g/L in the described milk-acid bacteria substratum.
The present invention preferably, described peptone is Tryptones and/or casein peptone.
The present invention preferably, described ammonium citrate is no more than 5g/L; Described dipotassium hydrogen phosphate is no more than 2.5g/L; Described sal epsom is no more than 0.3g/L; Described manganous sulfate is no more than 0.2g/L.
The present invention more preferably, described milk-acid bacteria substratum is made up of following component: Tryptones 10～12g/L, beef extract 9～10g/L, yeast extract 4～5g/L, glucose 20～22g/L, ammonium citrate 1.5～2.0g/L, dipotassium hydrogen phosphate 2.0～2.5g/L, sal epsom 0.2g/L, manganous sulfate 0.04～0.2g/L and agar 10～14g/L, the pH value of described substratum is 6.88-6.91.
Among the present invention, the medium that dissolves described milk-acid bacteria substratum is this area routine, preferably is water; Described water is that this area routine is described, as deionized water, distilled water, distilled water etc.
The preparation method of milk-acid bacteria substratum of the present invention is this area routine, only needs its each component that comprises is simply mixed, and the preparation condition preparation according to the conventional microbiological culture media in this area gets final product then.
Two of technical scheme provided by the invention is: foregoing milk-acid bacteria substratum has application in the inhibiting milk-acid bacteria in screening to helicobacter pylori.
On the basis that meets this area general knowledge, above-mentioned each optimum condition, but arbitrary combination namely get the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material be commercially available getting all.
Positive progressive effect of the present invention is:
Substratum of the present invention can well be grown for milk-acid bacteria, and having overcome in the past, nearly all business-like MRS substratum has inhibiting problem to helicobacter pylori, can be used for screening has inhibiting milk-acid bacteria to helicobacter pylori, obtains true and reliable The selection result in screening experiment; Simultaneously, substratum of the present invention greatly reduces the cost of milk-acid bacteria substratum.The present patent application people has found in the correlative study before from NM important phenomenon, be that milk-acid bacteria substratum itself exists restraining effect to helicobacter pylori, and use substratum of the present invention can get rid of interference in the milk-acid bacteria screening experiment, to there be the experiment of inhibiting milk-acid bacteria to bring basic change to helicobacter pylori for screening, thereby lay the foundation for correlative study from now on obtains real conclusion.
Description of drawings
Fig. 1 be the MRS substratum of Merck company and Oxoid company to the inhibiting figure as a result of helicobacter pylori, its left side is the substratum of Oxoid company, the right is the substratum of Merck company.
Fig. 2 is the comparing result figure of MRS substratum aspect the inhibition helicobacter pylori activity of substratum of the present invention and Merck company and Oxoid company, wherein, a is substratum of the present invention, and b is the MRS substratum of Oxoid company, and c is the MRS substratum of Merck company.
Embodiment
Mode below by embodiment further specifies the present invention, but does not therefore limit the present invention among the described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example according to ordinary method and condition, or is selected according to catalogue.
Among the following embodiment used medicine if no special instructions, all available from medicine company limited-liability company of traditional Chinese medicines group.Commercialization MRS substratum is available from Merck company and Oxoid company.
Embodiment 1
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is as follows:
Amount with 1L is prepared, and accurately takes by weighing above each component, adds distilled water to 1L, and heated and stirred is dissolved fully to component, and 121 ℃, stand-by after the 15min sterilising treatment, its final pH value is 6.91.
Embodiment 2
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 6.65.
Embodiment 3
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 6.88.
Embodiment 4
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 6.70.
Embodiment 5
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 5.84.
Embodiment 6
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 6.20.
Embodiment 7
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 6.05.
Embodiment 8
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 6.15.
Embodiment 9
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 6.55.
Embodiment 10
Each component that the milk-acid bacteria substratum of present embodiment comprises is as shown in the table:
Its preparation method is with embodiment 1, and its final pH value is 5.96.
Embodiment 11
Its preparation method is with embodiment 1, and its final pH value is 6.68.
The test of effect embodiment 1 lactobacter growth situation
With plant lactobacillus (Lb.plantarum) ST-III bacterial strain (CGMCC NO.0847, available from bright dairy industry research institute fundamental research portion, this bacterial strain information has disclosure in CN1467290A) with 1%(v/v) inoculum size insert respectively in commercial MRS substratum and the embodiment of the invention 3,4,9 and 11 the substratum (all removing agar powder wherein), 37 ℃ of anaerobism are cultivated 24h, by measuring pH value and the OD value of fermentation back nutrient solution, observe the growth situation.Outcome record is in following table.(annotate: agar powder only plays grumeleuse in substratum, therefore, the solid of same substratum and liquid form only differ agar powder.）
| Testing index | Commercialization MRS(Merck) | Embodiment 3 | Embodiment 4 | Embodiment 9 | Embodiment 11 |
| Initial pH value | 5.84 | 6.88 | 6.70 | 6.55 | 6.68 |
| Fermentation back pH value | 3.98 | 4.05 | 4.00 | 3.95 | 3.98 |
| Initial OD 600Value | 0.131 | 0.140 | 0.135 | 0.138 | 0.140 |
| Fermentation back OD 600Value | 2.530 | 2.152 | 2.078 | 2.057 | 2.086 |
As can be seen from the above table, the absorbancy rate of increase of fermentation back nutrient solution does not have significant difference; Measured above-mentioned each Lactic Acid from Fermentation Broth bacterium total number of bacterial colony simultaneously, all 10
8More than the cfu/mL.
Explanation thus, substratum of the present invention can be grown well for milk-acid bacteria.
Effect embodiment 2 indication bacteriostatic test plates
The preparation of indicator flat board: helicobacter pylori (Helicobacter pylori) (available from ATCC, ATCC NO:43504) thalline is suspended in the Columbia substratum (sheep blood serum of adding 10%), makes about 10
9The bacteria suspension of cfu/mL, dull and stereotyped, wait to solidify the back as the indication flat board.
With the MRS substratum of Merck company, Oxoid company and the embodiment of the invention 3,5,9,10 substratum, prepare, sterilize, fall dull and stereotyped respectively, after waiting to solidify, take out the circular substratum grumeleuse of diameter 5～15mm with punch tool, place the top of above-mentioned indicator substratum, under the condition that helicobacter pylori is suitable for growing, i.e. three gas incubator (O
2: CO
2: N
2=5:10:85,37 ℃) the middle 48～72h that cultivates, observe circular substratum grumeleuse to the inhibition situation of indicator.Wherein, the result of embodiment 3 as depicted in figs. 1 and 2.
The left side of Fig. 1 and the right are respectively the MRS substratum of Merck company and Oxoid company, can from figure, see, all occurred very significantly inhibition zone around both, illustrated that the MRS substratum itself of Merck company and Oxoid company has restraining effect for helicobacter pylori.
Fig. 2 is the substratum of the embodiment of the invention 3 and the comparing result figure of MRS substratum aspect the inhibition helicobacter pylori activity of Merck company and Oxoid company, wherein, a is the substratum of the embodiment of the invention 3, and b is the MRS substratum of Oxoid company, and c is the MRS substratum of Merck company.As can be seen from the figure, the MRS substratum of Merck company and Oxoid company all has restraining effect (inhibition zone occurring) for helicobacter pylori; And substratum of the present invention does not have restraining effect (not having inhibition zone) to helicobacter pylori.
Embodiment 5,9 is consistent with 10 experimental result with embodiment 3, all shows helicobacter pylori unrestraint effect (inhibition zone not occurring), and other embodiment also have the result suitable with embodiment 3 aspect antibacterial at flat board.
The all embodiment of the present invention all can reach and neither suppress helicobacter pylori, can guarantee the purpose of the good growth of milk-acid bacteria again.Above effect embodiment illustrates that substratum of the present invention can guarantee the good growth of milk-acid bacteria, again helicobacter pylori there is not restraining effect, thereby can guarantee when screening has inhibiting milk-acid bacteria to helicobacter pylori, its result's reliability has overcome the defective that traditional milk-acid bacteria substratum itself has restraining effect to bring to helicobacter pylori.
Claims (9)
1. a milk-acid bacteria substratum is characterized in that, described milk-acid bacteria substratum is made up of following component:
1) peptone 10～12g/L;
2) yeast extract 4～6g/L;
3) glucose 6～22g/L;
4) acid-base buffer agent 2～7g/L, described acid-base buffer agent are ammonium citrate and/or dipotassium hydrogen phosphate;
5) somatomedin 0.15～0.4g/L, described somatomedin is sal epsom and/or manganous sulfate;
6) beef extract 0～10g/L;
7) Zulkovsky starch 0～15g/L;
8) fructose 0～3g/L; With
9) agar 0～20g/L;
The pH value of described substratum is 5.8-6.9.
2. milk-acid bacteria substratum as claimed in claim 1 is characterized in that, contains beef extract 8～10g/L in the described milk-acid bacteria substratum.
3. milk-acid bacteria substratum as claimed in claim 1 is characterized in that, contains Zulkovsky starch 5～15g/L in the described milk-acid bacteria substratum.
4. milk-acid bacteria substratum as claimed in claim 1 is characterized in that, contains fructose 3g/L in the described milk-acid bacteria substratum.
5. milk-acid bacteria substratum as claimed in claim 1 is characterized in that, contains agar 10～20g/L in the described milk-acid bacteria substratum.
6. milk-acid bacteria substratum as claimed in claim 1 is characterized in that, described peptone is Tryptones and/or casein peptone.
7. milk-acid bacteria substratum as claimed in claim 1 is characterized in that, described ammonium citrate is no more than 5g/L; Described dipotassium hydrogen phosphate is no more than 2.5g/L; Described sal epsom is no more than 0.3g/L; Described manganous sulfate is no more than 0.2g/L.
8. milk-acid bacteria substratum as claimed in claim 7, it is characterized in that, described milk-acid bacteria substratum comprises following component: Tryptones 10～12g/L, beef extract 9～10g/L, yeast extract 4～5g/L, glucose 20～22g/L, ammonium citrate 1.5～2.0g/L, dipotassium hydrogen phosphate 2.0～2.5g/L, sal epsom 0.2g/L, manganous sulfate 0.04～0.2g/L and agar 10～14g/L, the pH value of described substratum is 6.88-6.91.
9. helicobacter pylori there is application in the inhibiting milk-acid bacteria as each described milk-acid bacteria substratum of claim 1～8 in screening.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013101479292A CN103194417A (en) | 2013-04-25 | 2013-04-25 | Lactic acid bacterium culture medium |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013101479292A CN103194417A (en) | 2013-04-25 | 2013-04-25 | Lactic acid bacterium culture medium |
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| CN103194417A true CN103194417A (en) | 2013-07-10 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105296591A (en) * | 2015-11-26 | 2016-02-03 | 佛山市海天(高明)调味食品有限公司 | Culture medium for detecting difficult cultivation type lactic acid bacteria in food and detection method thereof |
| CN106148420A (en) * | 2015-04-27 | 2016-11-23 | 牡丹江佰佳信生物科技有限公司 | A kind of fermentation process improving lactic acid bacteria metabolite bacteriostatic activity |
| CN110951652A (en) * | 2019-12-30 | 2020-04-03 | 光明乳业股份有限公司 | Selenium-enriched lactobacillus preparation and preparation method thereof |
| CN112940983A (en) * | 2021-03-31 | 2021-06-11 | 盐城维康生物科技有限公司 | Lactobacillus plantarum preparation capable of increasing anti-helicobacter pylori effect and preparation method thereof |
-
2013
- 2013-04-25 CN CN2013101479292A patent/CN103194417A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 苗均莅等: "MRS培养基对幽门螺杆菌具有抑制作用", 《微生物学杂志》 * |
| 苗均莅等: "改良MRS对消除培养基自身抑制幽门螺杆菌生长的研究", 《乳业科学与技术》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106148420A (en) * | 2015-04-27 | 2016-11-23 | 牡丹江佰佳信生物科技有限公司 | A kind of fermentation process improving lactic acid bacteria metabolite bacteriostatic activity |
| CN105296591A (en) * | 2015-11-26 | 2016-02-03 | 佛山市海天(高明)调味食品有限公司 | Culture medium for detecting difficult cultivation type lactic acid bacteria in food and detection method thereof |
| CN105296591B (en) * | 2015-11-26 | 2018-08-07 | 佛山市海天(高明)调味食品有限公司 | A kind of culture medium and detection method for detecting difficult cultivation type lactic acid bacteria in food |
| CN110951652A (en) * | 2019-12-30 | 2020-04-03 | 光明乳业股份有限公司 | Selenium-enriched lactobacillus preparation and preparation method thereof |
| CN112940983A (en) * | 2021-03-31 | 2021-06-11 | 盐城维康生物科技有限公司 | Lactobacillus plantarum preparation capable of increasing anti-helicobacter pylori effect and preparation method thereof |
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Application publication date: 20130710 |