CN111548941A - Method for activating freeze-dried strain - Google Patents
Method for activating freeze-dried strain Download PDFInfo
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- CN111548941A CN111548941A CN202010340847.XA CN202010340847A CN111548941A CN 111548941 A CN111548941 A CN 111548941A CN 202010340847 A CN202010340847 A CN 202010340847A CN 111548941 A CN111548941 A CN 111548941A
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- Prior art keywords
- freeze
- dried
- strain
- activation
- escherichia coli
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- 238000000034 method Methods 0.000 title claims abstract description 15
- 230000003213 activating effect Effects 0.000 title claims abstract description 6
- 239000001963 growth medium Substances 0.000 claims abstract description 21
- 230000004913 activation Effects 0.000 claims abstract description 19
- 241000228245 Aspergillus niger Species 0.000 claims abstract description 14
- 241000588724 Escherichia coli Species 0.000 claims abstract description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims abstract description 11
- 239000007787 solid Substances 0.000 claims abstract description 9
- 239000011780 sodium chloride Substances 0.000 claims abstract description 6
- 239000007864 aqueous solution Substances 0.000 claims abstract description 4
- 230000001580 bacterial effect Effects 0.000 claims description 20
- 239000000725 suspension Substances 0.000 claims description 14
- 238000012258 culturing Methods 0.000 claims description 6
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- 241000894007 species Species 0.000 claims description 2
- 239000002699 waste material Substances 0.000 abstract description 4
- 238000001514 detection method Methods 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- 239000000843 powder Substances 0.000 abstract 1
- 238000005086 pumping Methods 0.000 description 8
- 230000002159 abnormal effect Effects 0.000 description 6
- 238000009630 liquid culture Methods 0.000 description 6
- 239000002504 physiological saline solution Substances 0.000 description 6
- 238000004153 renaturation Methods 0.000 description 6
- 239000002609 medium Substances 0.000 description 4
- SFAYBQDGCKZKMH-UHFFFAOYSA-N BNCC Chemical compound BNCC SFAYBQDGCKZKMH-UHFFFAOYSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- C12N1/04—Preserving or maintaining viable microorganisms
-
- 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
- C12N1/14—Fungi; 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
- C12N1/20—Bacteria; Culture media therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A method for activating a freeze-dried strain, belonging to the technical field of microbial detection. In order to save the activation cost of the freeze-dried strain and avoid the waste of a culture medium, the invention provides a strain activation method, which takes a sodium chloride aqueous solution with the mass fraction of 0.8-0.9% as an activation culture solution, mixes the strain and the activation culture solution and then inoculates the mixture on a solid culture medium for culture. The invention can be used for activating freeze-dried powder of Escherichia coli and Aspergillus niger.
Description
Technical Field
The invention belongs to the technical field of microbial detection, and particularly relates to a method for activating a freeze-dried strain.
Background
Some of the freeze-dried strains purchased at present are specified in the strain activation instruction to be firstly dissolved and activated by using a special liquid culture medium and then cultured in a solid culture medium. Some of the commercial strains do not have a dedicated liquid medium attached to them. If the special liquid culture medium purchased or prepared by the user is not used for other purposes and is only used for strain activation, the rest culture medium has a shelf life and can cause waste when the rest culture medium is not used completely.
Disclosure of Invention
In order to save the activation cost of the freeze-dried strain and avoid the waste of a culture medium, the invention provides a strain activation method, which takes a sodium chloride aqueous solution with the mass fraction of 0.8-0.9% as an activation culture solution, mixes the freeze-dried strain and the activation culture solution and then inoculates the mixture on a solid culture medium for culture.
Further defined, the lyophilized species is lyophilized Escherichia coli (Escherichia coli) or lyophilized Aspergillus niger (Aspergillus niger).
Further defined, the freeze-dried Escherichia coli is cultured at 37 ℃ for 18-24 h.
Further defined, the freeze-dried aspergillus niger is cultured at the temperature of 28 ℃ for 5-7 days.
More particularly, the freeze-dried Escherichia coli is activated by the following specific method: mixing lyophilized Escherichia coli with activation culture solution to obtain bacterial suspension, inoculating to NA solid culture medium, coating, and culturing at 37 deg.C for 18-24 h.
More specifically, the freeze-dried aspergillus niger is activated by the following method: mixing the freeze-dried aspergillus niger strains with the activated culture solution to obtain a bacterial suspension, inoculating the bacterial suspension to a Martin solid culture medium, and culturing for 5-7 days at 28 ℃.
Advantageous effects
According to the invention, the existing liquid culture medium is replaced by 0.8-0.9% (by mass) of sodium chloride aqueous solution, the activity of the cultured strain can meet the use requirement, the strain activation cost is saved, and the waste of the residual culture medium is also avoided.
Detailed Description
Preparing a sodium chloride solution: preparing 0.8-0.9% normal saline from sodium chloride and tertiary water, sterilizing at 121 deg.C for 15 min, and using.
Lyophilized Escherichia coli as described in the examples below was purchased from BNCC under the designation 269342; the lyophilized Aspergillus niger was purchased from BNCC under the cat # 286403.
EXAMPLE 1 activation of lyophilized Escherichia coli.
50ml of physiological saline and NB liquid medium with a mass fraction of 0.85% were prepared in Erlenmeyer flasks, respectively. Respectively opening the safety cut-open pipes of the freeze-dried strains in the biological safety cabinet, sucking 0.5ml of physiological saline and pumping into one safety cut-open pipe, sucking 0.5ml of NB culture medium and pumping into the other safety cut-open pipe. After the bacterial suspension is fully dissolved, pumping the bacterial suspension back to a liquid culture medium, mixing uniformly, respectively sucking 200ul of bacterial suspension, pumping the bacterial suspension onto two NA plates, uniformly coating, and culturing at 37 ℃ for 24 hours. After culture, the bacterial colonies on the two NA plates are observed to have good renaturation and no abnormal morphology.
Example 2. activation of lyophilized Aspergillus niger strains.
50ml of physiological saline and Martin broth with a mass fraction of 0.85% were prepared in a Erlenmeyer flask. Respectively opening the safety cut-open tubes of the freeze-dried strains in the biological safety cabinet, sucking 0.5ml of physiological saline and pumping into one safety cut-open tube, and sucking 0.5ml of martin liquid culture medium and pumping into the other safety cut-open tube. After the bacterial suspension is fully dissolved, pumping the bacterial suspension back to a liquid culture medium, uniformly mixing, respectively sucking 200ul of bacterial suspension, pumping the bacterial suspension onto two Martin solid culture media, uniformly coating, and culturing at 28 ℃ for 5 d. After culture, the bacterial colonies on the two Martin plates are observed to have good renaturation and no abnormal morphology.
Example 3. example 1 was repeated, except that Escherichia coli was cultured at 37 ℃ for 18 hours in this example. After culture, the bacterial colonies on the two NA plates are observed to have good renaturation and no abnormal morphology.
Example 4. example 2 was repeated, with the difference from example 2 that the A.niger strains were cultured at 28 ℃ for 7 days. After culture, the bacterial colonies on the two Martin plates are observed to have good renaturation and no abnormal morphology.
Example 5. example 1 was repeated, except that in this example, 0.8% by mass of physiological saline was used as the activation medium, unlike example 1. After culture, the bacterial colonies on the two NA plates are observed to have good renaturation and no abnormal morphology.
Example 6. example 2 was repeated, except that in this example, 0.9% by mass of physiological saline was used as the activation medium, unlike example 2. After culture, the bacterial colonies on the two Martin plates are observed to have good renaturation and no abnormal morphology.
Claims (6)
1. A method for activating a freeze-dried strain is characterized in that a sodium chloride aqueous solution with the mass fraction of 0.8% -0.9% is used as an activation culture solution, and the freeze-dried strain and the activation culture solution are mixed and then inoculated onto a solid culture medium for culture.
2. The method according to claim 1, wherein the lyophilized species is lyophilized Escherichia coli (Escherichia coli) or lyophilized Aspergillus niger (Aspergillus niger).
3. The method according to claim 2, wherein the lyophilized Escherichia coli is cultured at 37 ℃ for 18 to 24 hours.
4. The method according to claim 2, wherein the freeze-dried Aspergillus niger is cultured at 28 ℃ for 5-7 days.
5. The method according to claim 3, wherein the lyophilized Escherichia coli is activated in a specific manner as follows: mixing lyophilized Escherichia coli with activation culture solution to obtain bacterial suspension, inoculating to NA solid culture medium, coating, and culturing at 37 deg.C for 18-24 h.
6. The method according to claim 4, wherein the freeze-dried Aspergillus niger is activated by the following specific method: mixing the freeze-dried aspergillus niger strains with the activated culture solution to obtain a bacterial suspension, inoculating the bacterial suspension to a Martin solid culture medium, and culturing for 5-7 days at 28 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010340847.XA CN111548941A (en) | 2020-04-26 | 2020-04-26 | Method for activating freeze-dried strain |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010340847.XA CN111548941A (en) | 2020-04-26 | 2020-04-26 | Method for activating freeze-dried strain |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111548941A true CN111548941A (en) | 2020-08-18 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010340847.XA Pending CN111548941A (en) | 2020-04-26 | 2020-04-26 | Method for activating freeze-dried strain |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111548941A (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102356913A (en) * | 2011-08-31 | 2012-02-22 | 哈尔滨北极人商贸有限公司 | Preparation method of probiotics fermented blueberry pulp powder |
| CN102356912A (en) * | 2011-11-02 | 2012-02-22 | 黑龙江省龙蛙粮油进出口有限公司 | Preparation method of probiotic fermented rice milk |
| CN102599335A (en) * | 2012-03-07 | 2012-07-25 | 哈尔滨市海澳斯生物科技开发有限公司 | Method for preparing compound microorganism fermented forage feed |
| CN102763726A (en) * | 2012-08-15 | 2012-11-07 | 张玉文 | Probiotics yoghourt powder and preparation method thereof |
| CN102907612A (en) * | 2012-11-09 | 2013-02-06 | 黑龙江省轻工科学研究院 | Preparation method for compound microorganism fermented maize gruel |
| CN103380874A (en) * | 2012-05-05 | 2013-11-06 | 黑龙江大荒春酒业有限公司 | Preparation method for composite probiotic-fermented jam |
| CN103652327A (en) * | 2013-12-10 | 2014-03-26 | 青岛琅琊台集团股份有限公司 | Biological feed produced by wastes |
| CN107712767A (en) * | 2017-09-25 | 2018-02-23 | 佛山科学技术学院 | A kind of preparation method of roasting-type fermentation jam |
| CN111647538A (en) * | 2020-06-23 | 2020-09-11 | 科兴生物制药股份有限公司 | Clostridium butyricum freeze-dried powder, preparation method and application thereof |
-
2020
- 2020-04-26 CN CN202010340847.XA patent/CN111548941A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102356913A (en) * | 2011-08-31 | 2012-02-22 | 哈尔滨北极人商贸有限公司 | Preparation method of probiotics fermented blueberry pulp powder |
| CN102356912A (en) * | 2011-11-02 | 2012-02-22 | 黑龙江省龙蛙粮油进出口有限公司 | Preparation method of probiotic fermented rice milk |
| CN102599335A (en) * | 2012-03-07 | 2012-07-25 | 哈尔滨市海澳斯生物科技开发有限公司 | Method for preparing compound microorganism fermented forage feed |
| CN103380874A (en) * | 2012-05-05 | 2013-11-06 | 黑龙江大荒春酒业有限公司 | Preparation method for composite probiotic-fermented jam |
| CN102763726A (en) * | 2012-08-15 | 2012-11-07 | 张玉文 | Probiotics yoghourt powder and preparation method thereof |
| CN102907612A (en) * | 2012-11-09 | 2013-02-06 | 黑龙江省轻工科学研究院 | Preparation method for compound microorganism fermented maize gruel |
| CN103652327A (en) * | 2013-12-10 | 2014-03-26 | 青岛琅琊台集团股份有限公司 | Biological feed produced by wastes |
| CN107712767A (en) * | 2017-09-25 | 2018-02-23 | 佛山科学技术学院 | A kind of preparation method of roasting-type fermentation jam |
| CN111647538A (en) * | 2020-06-23 | 2020-09-11 | 科兴生物制药股份有限公司 | Clostridium butyricum freeze-dried powder, preparation method and application thereof |
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Application publication date: 20200818 |