CN106148192A - A kind of preparation method of antibacterial peptide strain seed liquor - Google Patents
A kind of preparation method of antibacterial peptide strain seed liquor Download PDFInfo
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- CN106148192A CN106148192A CN201610764007.XA CN201610764007A CN106148192A CN 106148192 A CN106148192 A CN 106148192A CN 201610764007 A CN201610764007 A CN 201610764007A CN 106148192 A CN106148192 A CN 106148192A
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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
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Abstract
The preparation method that the present invention relates to a kind of antibacterial peptide strain seed liquor comprises the following steps: the most aseptically, scrapes lower inclined plane mycelium in seed bottle with Inoculating needle, puts and cultivates 12 14 hours on 32 ± 1 DEG C of shaking tables；Then under aseptic condition, shake-flask seed is accessed in first class seed pot and cultivate, cultivation temperature 34 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 12 14 hours；Then first order seed is moved in secondary seed tank and cultivate, cultivation temperature 36 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 10 12 hours, obtain antibacterial peptide strain seed liquor；The volume of secondary seed tank is 5 10 times of first order seed tank volume.The method uses two steps to expand, and adds the volume of secondary seed tank, have adjusted mixing speed, cultivation temperature and incubation time simultaneously, obtains the effect of original three grades of amplification culture, it is to avoid occur, in transfer process, the situation that seed liquor is polluted.
Description
Technical field
The invention belongs to technical field of biological fermentation, be specifically related to the preparation method of a kind of antibacterial peptide strain seed liquor.
Background technology
Antibacterial peptide is that the interior one produced through induction of organism has bioactive micromolecule polypeptide, this kind of active polypeptide
Majority has the features such as alkalescence, heat stability and broad-spectrum antiseptic.Compared with antibiotic, to humans and animals normal cell without wound
Evil, has no side effect, noresidue, has wide in medicinal antibiosis field, food antiseptic preservation field and feed additive field
Wealthy application prospect.
But, at present a lot of problem is existed in commercial process for antibacterial peptide fermentation medium, as cost is high,
Antibacterial peptide strain seed liquor preparation process generally requires through three grades of amplification culture, during passing on, there will be seed liquor
Situation about polluting, affects the production performance of strain, and then governs yield and the quality of antibacterial peptide.
Summary of the invention
Object of the present invention is to provide the preparation method of a kind of antibacterial peptide strain seed liquor, the method uses two steps to expand
Greatly, add the volume of secondary seed tank, have adjusted mixing speed, cultivation temperature and incubation time simultaneously, obtain original three grades
The effect of amplification culture, and then improve the productivity of antibacterial peptide, it is to avoid in transfer process, the situation that seed liquor is polluted occurs.
For achieving the above object, the technical solution used in the present invention is as follows:
The invention provides the preparation method of a kind of antibacterial peptide strain seed liquor, it comprises the following steps:
(1) preparation of shake-flask seed: shake-flask seed culture medium, after sterilizing cools down, aseptically, scrapes with Inoculating needle
Inclined-plane mycelium, in seed bottle, is put after cultivating 12-14 hour on 32 ± 1 DEG C of shaking tables, and lower shaking table is standby；
(2) preparation of first order seed: shake-flask seed, after sterilizing cools down, is aseptically accessed by primary-seed medium
First class seed pot is cultivated, cultivation temperature 34 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 12-14 hour；
(3) preparation of secondary seed: secondary seed medium is after sterilizing cools down, by the subcultivation pipeline of sterilizing by
Level seed moves in secondary seed tank to be cultivated, and cultivation temperature 36 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 10-12 is little
Time, obtain antibacterial peptide strain seed liquor；5-10 times that volume is first order seed tank volume of described secondary seed tank.
According to the preparation method of above-mentioned antibacterial peptide strain seed liquor, the cultivation temperature in step (3) is than in step (2)
Cultivation temperature height 1-2 DEG C.
According to the preparation method of above-mentioned antibacterial peptide strain seed liquor, the mixing speed of the secondary seed tank in step (3)
10-20rpm is improved than the mixing speed in the first class seed pot in step (2).
Compared with prior art, beneficial effects of the present invention:
The inventive method uses two steps to expand, and adds the volume of secondary seed tank, have adjusted mixing speed, cultivation temperature simultaneously
And incubation time, obtain the effect of original three grades of amplification culture, and then improve the productivity of antibacterial peptide, it is to avoid in transfer process
The situation that seed liquor is polluted occurs.
Detailed description of the invention
Following example are intended to further illustrate present disclosure, but are not intended to protection scope of the present invention.
Embodiment 1
A kind of preparation method of antibacterial peptide strain seed liquor, it comprises the following steps:
(1) preparation of shake-flask seed: shake-flask seed culture medium, after sterilizing cools down, aseptically, scrapes with Inoculating needle
Inclined-plane mycelium, in seed bottle, is put after cultivating 12-14 hour on 32 ± 1 DEG C of shaking tables, and lower shaking table is standby；
(2) preparation of first order seed: shake-flask seed, after sterilizing cools down, is aseptically accessed by primary-seed medium
First class seed pot is cultivated, cultivation temperature 34 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 12-14 hour；
(3) preparation of secondary seed: secondary seed medium is after sterilizing cools down, by the subcultivation pipeline of sterilizing by
Level seed moves in secondary seed tank to be cultivated, and cultivation temperature 36 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 10-12 is little
Time, obtain antibacterial peptide strain seed liquor；5-10 times that volume is first order seed tank volume of described secondary seed tank.
In the preparation method of above-mentioned antibacterial peptide strain seed liquor, the cultivation temperature in step (3) is than the training in step (2)
Support temperature height 1-2 DEG C；The mixing speed of the secondary seed tank in step (3) is than the stirring in the first class seed pot in step (2)
Speed improves 10-20rpm.
Under equal conditions, utilize the inventive method to prepare compared with traditional third stage culture method, prepared by the inventive method
The performance of antibacterial peptide strain seed liquor prepared by the performance of antibacterial peptide strain seed liquor and traditional method is suitable.
Claims (3)
1. a preparation method for antibacterial peptide strain seed liquor, it comprises the following steps:
(1) preparation of shake-flask seed: shake-flask seed culture medium, after sterilizing cools down, aseptically, scrapes with Inoculating needle
Inclined-plane mycelium, in seed bottle, is put after cultivating 12-14 hour on 32 ± 1 DEG C of shaking tables, and lower shaking table is standby；
(2) preparation of first order seed: shake-flask seed, after sterilizing cools down, is aseptically accessed by primary-seed medium
First class seed pot is cultivated, cultivation temperature 34 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 12-14 hour；
(3) preparation of secondary seed: secondary seed medium is after sterilizing cools down, by the subcultivation pipeline of sterilizing by
Level seed moves in secondary seed tank to be cultivated, and cultivation temperature 36 ± 1 DEG C, ventilation 1:1 v/v/min, incubation time 10-12 is little
Time, obtain antibacterial peptide strain seed liquor；5-10 times that volume is first order seed tank volume of described secondary seed tank.
The preparation method of antibacterial peptide strain seed liquor the most according to claim 1, it is characterised in that: the training in step (3)
Support temperature than cultivation temperature height 1-2 DEG C in step (2).
The preparation method of antibacterial peptide strain seed liquor the most according to claim 1, it is characterised in that: two in step (3)
The mixing speed of level seed tank improves 10-20rpm than the mixing speed in the first class seed pot in step (2).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610764007.XA CN106148192A (en) | 2016-08-30 | 2016-08-30 | A kind of preparation method of antibacterial peptide strain seed liquor |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610764007.XA CN106148192A (en) | 2016-08-30 | 2016-08-30 | A kind of preparation method of antibacterial peptide strain seed liquor |
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| CN106148192A true CN106148192A (en) | 2016-11-23 |
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| CN201610764007.XA Pending CN106148192A (en) | 2016-08-30 | 2016-08-30 | A kind of preparation method of antibacterial peptide strain seed liquor |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102505032A (en) * | 2011-11-03 | 2012-06-20 | 福建省建阳武夷味精有限公司 | Method for supplementing seed liquid in sisomicin fermentation process |
| CN103194411A (en) * | 2013-04-08 | 2013-07-10 | 中国农业大学 | Bacillus subtilis and application of bacillus subtilis to production of antimicrobial peptide |
| CN103849667A (en) * | 2012-11-30 | 2014-06-11 | 中农颖泰林州生物科园有限公司 | Cecropin antibacterial peptide production process |
| CN103966297A (en) * | 2014-05-30 | 2014-08-06 | 山东仙普爱瑞科技股份有限公司 | Fermentation process for producing antibacterial peptides from bacillus subtilis |
-
2016
- 2016-08-30 CN CN201610764007.XA patent/CN106148192A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102505032A (en) * | 2011-11-03 | 2012-06-20 | 福建省建阳武夷味精有限公司 | Method for supplementing seed liquid in sisomicin fermentation process |
| CN103849667A (en) * | 2012-11-30 | 2014-06-11 | 中农颖泰林州生物科园有限公司 | Cecropin antibacterial peptide production process |
| CN103194411A (en) * | 2013-04-08 | 2013-07-10 | 中国农业大学 | Bacillus subtilis and application of bacillus subtilis to production of antimicrobial peptide |
| CN103966297A (en) * | 2014-05-30 | 2014-08-06 | 山东仙普爱瑞科技股份有限公司 | Fermentation process for producing antibacterial peptides from bacillus subtilis |
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Application publication date: 20161123 |
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