CN105779354A - Escherichia coli culture medium - Google Patents
Escherichia coli culture medium Download PDFInfo
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- CN105779354A CN105779354A CN201610235321.9A CN201610235321A CN105779354A CN 105779354 A CN105779354 A CN 105779354A CN 201610235321 A CN201610235321 A CN 201610235321A CN 105779354 A CN105779354 A CN 105779354A
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- escherichia coli
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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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Abstract
The invention relates to an escherichia coli culture medium.Each liter of the culture medium is prepared from, by weight, 10-20 g of glucose, 10 g of sodium nitrate, 5-14 g of peptone, 0.2-3 g of monopotassium phosphate and 0.5-5 g of dipotassium phosphate.A preparation method of the novel escherichia coli culture medium includes the following steps that 1, glucose, sodium nitrate, peptone, monopotassium phosphate and dipotassium phosphate are added into a vessel, water is added, the materials are stirred to be dissolved, and the volume is made constant; 2, the pH value is regulated; 3, the culture medium is sub-packaged; 4, sterilization is carried out.The escherichia coli culture medium is economical and practical and can be prepared in large batches; the growth speed of escherichia coli can be regulated by changing the concentration of components in the culture medium.
Description
Technical field
The present invention relates to biological technical field, particularly relate to a kind of Escherichia coli culture medium and preparation method thereof.
Background technology
Escherichia coli (formal name used at school: Escherichia coli, be generally abbreviated as E.coli)) it is to live in homoiothermic animal
Significant bacterial kind in (including birds and mammal) large intestine, has important work to food normal stool
With.Escherichia coli are a member of enterobacteriaceae, and due to its simple in construction, genome is sequenced, often makees
Model organism for antibacterial is widely used in scientific research.Especially with the hands such as Physiology and biochemistry and molecular biology
Section has greatly promoted the parsing to other biological life mechanism to the research of escherichia coli vital functions.Additionally,
In biological engineering, escherichia coli are widely used in the host of gene replication and expression.
At Water warfare and sewage treatment area, escherichia coli are selected as the indicative thing of water pollution degree very early
Kind.At present, escherichia coli have been widely used as the indicator bacteria of food hygiene quality inspection.The food of coliform
Product health significance is the indicator bacteria as food faecal contamination, as long as manure content reaches in food
10-3mg/kg can detect escherichia coli.
Culture medium (Medium) is manually to be configured to by the needs of growth of microorganism metabolism by multiple nutrients material
A kind of nutrient matrix, be the material base in indoors artificial cultivating microorganism.And microorganism is in the medium
Growth be the indoor prerequisite that microorganism is carried out scientific research, as the microorganism in solid medium can
Being used for separating, identifying and conservation, the microorganism of liquid culture then can be utilized for the research of metabolite.To the greatest extent
Pipe is different with research purpose due to the kind of microorganism, each variant in the preparation of culture medium constituent,
But the preparation of any culture medium be both needed to meet growth of microorganism grow necessary moisture, carbon source, nitrogen source,
Inorganic salt, somatomedin and some trace element etc. special procured.Additionally, also need during culture medium preparation to consider
Equilibrium between each nutritional labeling and coordination, and suitable acid-base value (pH value), buffer capacity, oxidation
Reduction potential and osmotic pressure etc..
At present, conventional Escherichia coli culture medium has LB culture medium, beef-protein medium, cholate breast
Sugar culture-medium etc..Mostly there is preparation high cost in existing culture medium, investment is big, be difficult to high-volume prepares
Problem.Especially indoor cultivation test in, often because domestic medicine purity is inadequate, impurity is more from
Abroad purchase medicine, further increase culture medium preparation cost.Although these culture medium are widely used,
If but to carry out the lab simulation of the aspects such as the water body purification big to culture medium demand and sewage disposal in fact
Test, can high by cost, invest big restriction, and then experiment repetitive rate can be brought low and experiment effect difference etc.
Problem.
In view of drawbacks described above, the design people's the most in addition research and innovation, cultivate to founding a kind of escherichia coli
Base so that it is have more the value in industry.
Summary of the invention
For solving above-mentioned technical problem, it is an object of the invention to provide a kind of Escherichia coli culture medium, this cultivation
Base be more economical, more practical than LB culture medium, beef-protein medium, cholate lactose medium etc. and
And equivalence, the high-volume low cost preparation of culture medium can be realized.
On the one hand, the present invention provides a kind of Escherichia coli culture medium, containing following weight in every liter of culture medium
Each component: glucose 10-20g, sodium nitrate 10g, peptone 5-14g, potassium dihydrogen phosphate 0.2-3g, phosphorus
Acid hydrogen dipotassium 0.5-5g, surplus is water.
In one embodiment, containing each component of following weight in every liter of culture medium: glucose 10g, nitre
Acid sodium 10g, peptone 5g, potassium dihydrogen phosphate 0.2g, dipotassium hydrogen phosphate 0.8g, surplus is water.
In another specific embodiment, every liter of culture medium contains each component of following weight: glucose 15g, nitre
Acid sodium 10g, peptone 10g, potassium dihydrogen phosphate 1.5g, dipotassium hydrogen phosphate 3g, surplus is water.
In still another embodiment, every liter of culture medium contains each component of following weight: glucose 20g, nitre
Acid sodium 10g, peptone 14g, potassium dihydrogen phosphate 3g, dipotassium hydrogen phosphate 5g, surplus is water.
On the other hand, the present invention also provides for the preparation method of above-mentioned Escherichia coli culture medium, including following step
Rapid:
(1) glucose, sodium nitrate, peptone, potassium dihydrogen phosphate, dipotassium hydrogen phosphate are joined in container,
Add water and stirring and dissolving, constant volume；
(2) regulation pH value；
(3) subpackage culture medium；
(4) sterilizing.
Further, in step (1), water is sterilized water.
Further, in step (2), by pH value regulation to 7.2-7.4.
Further, in step (3), the dispensed loading amount of culture medium is to be advisable less than the 1/2 of conical flask.
Further, in step (4), sterilizing under 0.025-0.1MPa pressure.
Further, in step (4), when concentration of glucose is more than 15g/L, 105-115 DEG C of temperature
The lower sterilizing of degree；When concentration of glucose is less than 15g/L, sterilizing at a temperature of 115-121 DEG C.
Further, in step (4), sterilization time is 20-30 minute.
Further, in step (4), when temperature is 116-121 DEG C, sterilizing 20 minutes, temperature is
When 105-115 DEG C, sterilizing 30 minutes.
Further, in step (1), change glucose, peptone, potassium dihydrogen phosphate, phosphoric acid hydrogen two
The amount of potassium can regulate the colibacillary speed of growth.
By such scheme, the beneficial effects of the present invention is:
(1) the invention provides a kind of Escherichia coli culture medium, select glucose, sodium nitrate, biphosphate
Potassium, dipotassium hydrogen phosphate provide carbon source, nitrogen source, phosphate for Escherichia coli Growth, replace yeast extract,
The high price reagent such as Carnis Bovis seu Bubali cream, reduce cost and put into；
(2) present invention selects peptone to provide nitrogen source, carbon source, somatomedin, dimension life for Escherichia coli Growth
Elements etc., replace high price reagent tryptone, reduce cost and put into；
(3) present invention selects sodium nitrate on the one hand to provide nitrogen source, on the other hand Na to Escherichia coli Growth+Energy
Maintain Premeabilisation of cells pressure；
(4) present invention is by changing the agent of the amount of glucose, peptone, potassium dihydrogen phosphate, dipotassium hydrogen phosphate
Amount, can regulate and control the speed of Escherichia coli Growth.
Therefore, the invention provides one than LB culture medium, beef-protein medium, the training of cholate lactose
Support that base etc. is more economical, practical and the Escherichia coli culture medium of equivalence, will can realize the high-volume of culture medium
Preparation；Can also be by changing the speed of the concentration regulation Escherichia coli Growth of component in culture medium.
Described above is only the general introduction of technical solution of the present invention, in order to better understand the technology of the present invention
Means, below in conjunction with specific embodiment, the invention will be further described.Following example are used for illustrating this
Invention, but it is not limited to the scope of the present invention.
Accompanying drawing explanation
Fig. 1 is colibacillary growth curve in 1# culture medium；
Fig. 2 is colibacillary growth curve in 2# culture medium；
Fig. 3 is colibacillary growth curve in 3# culture medium；
Fig. 4 is 1#, 2#, 3# culture medium and colibacillary growth curve comparison diagram in LB culture medium；
Detailed description of the invention
Below in conjunction with the accompanying drawings and embodiment, the detailed description of the invention of the present invention is described in further detail.With
Lower embodiment is used for illustrating the present invention, but is not limited to the scope of the present invention.
Embodiment 1
1, the preparation process of escherichia coli 1# culture medium:
Weigh glucose 10g, sodium nitrate 10g, peptone 5g, potassium dihydrogen phosphate 0.2g, dipotassium hydrogen phosphate
0.8g adds in beaker, is subsequently adding a small amount of sterilized water, is completely dissolved to medicine with Glass rod stirring, proceeds to
The volumetric flask of 1L adds sterilized water constant volume；The pH value of regulation solution is 7.2；By each for culture medium solution point
In the conical flask of 250ml to four 500ml of dress, after subpackage is complete, sealing of jumping a queue；At 121 DEG C, 0.1MPa
Sterilizing 20 minutes under pressure.
2, in the colibacillary step of 1# inoculation of medium:
(1) on conical flask, post label before inoculation, indicate bacterium name, inoculation date, inoculation people's name.
(2) alcohol burner is lighted.
(3) inoculation, with aseptic pipet by strain transposing to the conical flask posting label, operation must
Must carry out by aseptic manipulation, specifically comprise the following steps that
1. tampon is unscrewed, in order to extract during inoculation；
2. take Sterile pipette, draw 2ml Escherichia coli bacteria liquid；
3. pull out tampon, then allow conical flask mouth slowly overdo sterilizing (being sure not to burn boiling the hottest)；
4. the bacterium solution in pipet is connected in culture medium；
5. fill in tampon, do not go to meet tampon with conical flask, in order to avoid flask includes unclean air in when mobile；
6., under the conditions of 37 DEG C, cultivate 24 hours in microorganism shaking table under 200rpm rotating speed, obtain large intestine
Bacillus culture fluid.
3, escherichia coli growth curve in 1# culture medium measures:
Utilize spectrophotometric determination E. coli broth OD600 value.With OD600 value as vertical coordinate, time
Between for abscissa map, draw colibacillary growth curve, as shown in Figure 1.
Embodiment 2
1, the preparation process of escherichia coli 2# culture medium:
Weigh glucose 15g, sodium nitrate 10g, peptone 10g, potassium dihydrogen phosphate 1.5g, dipotassium hydrogen phosphate
3g adds in beaker, is subsequently adding a small amount of sterilized water, is completely dissolved to medicine with Glass rod stirring, proceeds to 1L
Volumetric flask in add sterilized water constant volume；The pH value of regulation solution is 7.3；By each for culture medium solution subpackage
In the conical flask of 250ml to four 500ml；At 115 DEG C, sterilizing 30 minutes under 0.075MPa pressure.
2, in the colibacillary step of 2# inoculation of medium:
(1) on conical flask, post label before inoculation, indicate bacterium name, inoculation date, inoculation people's name.
(2) alcohol burner is lighted.
(3) inoculation, with aseptic pipet by strain transposing to the conical flask posting label, operation must
Must carry out by aseptic manipulation, specifically comprise the following steps that
1. tampon is unscrewed, in order to extract during inoculation；
2. take Sterile pipette, draw 2ml Escherichia coli bacteria liquid；
3. pull out tampon, then allow conical flask mouth slowly overdo sterilizing (being sure not to burn boiling the hottest)；
4. the bacterium solution in pipet is connected in culture medium；
5. fill in tampon, do not go to meet tampon with conical flask, in order to avoid flask includes unclean air in when mobile；
6., under the conditions of 37 DEG C, cultivate 24 hours in microorganism shaking table under 200rpm rotating speed, obtain large intestine
Bacillus culture fluid.
3, escherichia coli growth curve in 2# culture medium measures
Utilize spectrophotometric determination E. coli broth OD600 value.With OD600 value as vertical coordinate, time
Between for abscissa map, draw colibacillary growth curve, as shown in Figure 2.
Embodiment 3
1, the preparation process of escherichia coli 3# culture medium:
Weigh glucose 20g, sodium nitrate 10g, peptone 14g, potassium dihydrogen phosphate 3g, dipotassium hydrogen phosphate 5g
Add in beaker, be subsequently adding a small amount of sterilized water, be completely dissolved to medicine with Glass rod stirring, proceed to 1L's
Volumetric flask adds sterilized water constant volume；The pH value of regulation solution is 7.4；By each for culture medium solution subpackage
In the conical flask of 250ml to four 500ml；At 105 DEG C, sterilizing 30 minutes under 0.025MPa pressure.
2, in the colibacillary step of 3# inoculation of medium:
(1) on conical flask, post label before inoculation, indicate bacterium name, inoculation date, inoculation people's name.
(2) alcohol burner is lighted.
(3) inoculation, with aseptic pipet by strain transposing to the conical flask posting label, operation must
Must carry out by aseptic manipulation, specifically comprise the following steps that
1. tampon is unscrewed, in order to extract during inoculation；
2. take Sterile pipette, draw 2ml Escherichia coli bacteria liquid；
3. pull out tampon, then allow conical flask mouth slowly overdo sterilizing (being sure not to burn boiling the hottest)；
4. the bacterium solution in pipet is connected in culture medium；
5. fill in tampon, do not go to meet tampon with conical flask, in order to avoid flask includes unclean air in when mobile；
6., under the conditions of 37 DEG C, cultivate 24 hours in microorganism shaking table under 200rpm rotating speed, obtain large intestine
Bacillus culture fluid.
3, escherichia coli growth curve in 3# culture medium measures:
Utilize spectrophotometric determination E. coli broth OD600 value.With OD600 value as vertical coordinate, time
Between for abscissa map, draw colibacillary growth curve, as shown in Figure 3.
Table 1 is the price detail list of the Escherichia coli culture medium of LB culture medium and the present invention.Can be seen by table 1
Going out, LB culture medium cost is tens times of the Escherichia coli culture medium that the present invention is prepared, and the present invention drops significantly
The low deployment cost of culture medium.
Table 1
As seen from Figure 4, the concentration of material in culture medium is changed, thus it is possible to vary colibacillary growth speed
Degree；And colibacillary growth curve and the growth curve in LB culture medium in 1#, 2#, 3# culture medium
Basic simlarity, illustrates that the Escherichia coli culture medium that the present invention provides is than traditional LB culture medium economy and equivalent.
Therefore, the invention provides one than LB culture medium, beef-protein medium, the training of cholate lactose
Support that base etc. is more economical, practical and the Escherichia coli culture medium of equivalence, will can realize the high-volume of culture medium
Preparation；Can also be by changing the speed of the concentration regulation Escherichia coli Growth of component in culture medium.
The above is only the preferred embodiment of the present invention, is not limited to the present invention.It should be pointed out that,
For those skilled in the art, on the premise of without departing from the technology of the present invention principle, also
Can make some improvement and modification, these improve and modification also should be regarded as protection scope of the present invention.
Claims (10)
1. an Escherichia coli culture medium, it is characterised in that: every liter of culture medium contains each group of following weight
Point: glucose 10-20g, sodium nitrate 10g, peptone 5-14g, potassium dihydrogen phosphate 0.2-3g, phosphoric acid hydrogen two
Potassium 0.5-5g, surplus is water.
Escherichia coli culture medium the most according to claim 1, it is characterised in that: every liter of culture medium contains
There is each component of following weight: glucose 10g, sodium nitrate 10g, peptone 5g, potassium dihydrogen phosphate 0.2g,
Dipotassium hydrogen phosphate 0.8g, surplus is water.
Escherichia coli culture medium the most according to claim 1, it is characterised in that: every liter of culture medium contains
There is each component of following weight: glucose 15g, sodium nitrate 10g, peptone 10g, potassium dihydrogen phosphate 1.5g,
Dipotassium hydrogen phosphate 3g, surplus is water.
Escherichia coli culture medium the most according to claim 1, it is characterised in that: every liter of culture medium contains
There is each component of following weight: glucose 20g, sodium nitrate 10g, peptone 14g, potassium dihydrogen phosphate 3g,
Dipotassium hydrogen phosphate 5g, surplus is water.
5. a preparation method for the Escherichia coli culture medium according to any one of claim 1-4, including with
Lower step:
(1) glucose, sodium nitrate, peptone, potassium dihydrogen phosphate, dipotassium hydrogen phosphate are joined in container,
Add water and stirring and dissolving, constant volume；
(2) regulation pH value；
(3) subpackage culture medium；
(4) sterilizing.
The preparation method of Escherichia coli culture medium the most according to claim 5, it is characterised in that: in step
Suddenly, in (1), described water is sterilized water.
The preparation method of Escherichia coli culture medium the most according to claim 5, it is characterised in that: in step
Suddenly in (2), by pH value regulation to 7.2-7.4.
The preparation method of Escherichia coli culture medium the most according to claim 5, it is characterised in that: in step
Suddenly in (4), sterilizing under 0.025-0.1MPa pressure.
The preparation method of Escherichia coli culture medium the most according to claim 5, it is characterised in that: in step
Suddenly in (4), when concentration of glucose is more than 15g/L, sterilizing at 105-115 DEG C；Work as concentration of glucose
During less than 15g/L, sterilizing at a temperature of 115-121 DEG C.
The preparation method of Escherichia coli culture medium the most according to claim 5, it is characterised in that:
In step (4), sterilization time is 20-30 minute.
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| CN201610235321.9A CN105779354A (en) | 2016-04-15 | 2016-04-15 | Escherichia coli culture medium |
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| CN201610235321.9A CN105779354A (en) | 2016-04-15 | 2016-04-15 | Escherichia coli culture medium |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434426A (en) * | 2016-08-30 | 2017-02-22 | 内蒙古金源康生物工程有限公司 | High-efficiency expression culture medium for escherichia coli |
| CN106754843A (en) * | 2016-12-14 | 2017-05-31 | 曹书华 | A kind of method for improving recombination bacillus coli bile salt hydrolase yield |
| CN111849854A (en) * | 2020-07-24 | 2020-10-30 | 暨南大学 | Method for enhancing mechanical strength of surface of escherichia coli |
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| CN103667137A (en) * | 2013-12-10 | 2014-03-26 | 中山奈德生物科技有限公司 | Escherichia coli culture medium and culturing method of escherichia coli culture medium |
| CN104877939A (en) * | 2015-05-27 | 2015-09-02 | 成都易创思生物科技有限公司 | Culture medium and cultural method applied to escherichia coli |
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2016
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Patent Citations (3)
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| CN102634502A (en) * | 2012-03-19 | 2012-08-15 | 苏州汉酶生物技术有限公司 | Production method of halide alcohol dehalogenase |
| CN103667137A (en) * | 2013-12-10 | 2014-03-26 | 中山奈德生物科技有限公司 | Escherichia coli culture medium and culturing method of escherichia coli culture medium |
| CN104877939A (en) * | 2015-05-27 | 2015-09-02 | 成都易创思生物科技有限公司 | Culture medium and cultural method applied to escherichia coli |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434426A (en) * | 2016-08-30 | 2017-02-22 | 内蒙古金源康生物工程有限公司 | High-efficiency expression culture medium for escherichia coli |
| CN106754843A (en) * | 2016-12-14 | 2017-05-31 | 曹书华 | A kind of method for improving recombination bacillus coli bile salt hydrolase yield |
| CN111849854A (en) * | 2020-07-24 | 2020-10-30 | 暨南大学 | Method for enhancing mechanical strength of surface of escherichia coli |
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Application publication date: 20160720 |