CN109603212A - A kind of microbial fermentation solution rapid sedimentation method - Google Patents
A kind of microbial fermentation solution rapid sedimentation method Download PDFInfo
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- CN109603212A CN109603212A CN201811530982.XA CN201811530982A CN109603212A CN 109603212 A CN109603212 A CN 109603212A CN 201811530982 A CN201811530982 A CN 201811530982A CN 109603212 A CN109603212 A CN 109603212A
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- Prior art keywords
- microbial fermentation
- fermentation solution
- sedimentation method
- rapid sedimentation
- calcium salt
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D21/00—Separation of suspended solid particles from liquids by sedimentation
- B01D21/01—Separation of suspended solid particles from liquids by sedimentation using flocculating agents
Abstract
A kind of microbial fermentation solution rapid sedimentation method, comprising the following steps: microbial fermentation solution is fitted into container by S1；Dibasic alkaliine and calcium salt mixed liquor are added into container by S2, and the mixing quality of dibasic alkaliine and calcium salt accounts for the 5%~10% of quality of fermentation broth；S3 shakes up above-mentioned solution or is stirred evenly with sterile glass rod, after standing, is placed in 4 DEG C of environment 12~24 hours, sops up supernatant, collects the thallus that precipitating is concentration.The present invention has the advantage that: it is non-toxic efficient, low in cost, can be by 50-68 times of concentration in microbial fermentation solution, cell yield is up to 90%.
Description
Technical field
The present invention relates to microorganisms technical field, in particular to a kind of microbial fermentation solution rapid sedimentation method.
Background technique
Microbial bacterial agent refers to that objective microbe (effective bacterium) after industrialized production expansion is numerous, utilizes porous substance
As adsorbent (such as turf, vermiculite), the active bacteria formulation that the fermentation liquid of thallus is processed into is adsorbed.This microbial inoculum is for dressing seed
Or root dipping, have directly or indirectly improvement soil, restore soil fertility, prevention soil-borne disease, maintain rhizosphere microflora balance and
The effects of degradation poisonous and harmful substances.Agricultural microbial agent appropriately using can be improved agricultural output, improve agricultural product product
Matter reduces fertilizer amount, reduces cost, improvement soil, preserves the ecological environment.
The features such as since it is with having no toxic side effect, noresidue is polluted, and does not generate resistance, in aquaculture, organic waste
Object compost fermentation and soil ecology reparation etc. play an important role, and microbial fermentation liquid precipitate and concentration can press significantly
Contracting storage volume reduces transportation cost, improves using effect.Therefore, people often take bacterium solution different modes different degrees of
Concentration；At present there are mainly two types of the method for concentration of microorganism formulation: first is that physical method, including centrifugal process and filtration method, this
Kind method for concentration equipment investment is high, and operating cost is also high；Second is that chemical method: mainly using high scores such as alum, poly-aluminium, Polyferric Sulfates
Sub- compound has toxicity as flocculant, these flocculants itself, to the toxic effect of microniological proudcts, while to micro- life
Produce product are also adversely affected using object and its environment.Therefore, a kind of preferably concentration is developed for microbial bacterial agent
Method is that those skilled in the art have one of problem to be solved.
Summary of the invention
It is non-toxic efficient, at low cost it is an object of the invention to provide a kind of microbial fermentation solution rapid sedimentation method
It is honest and clean, can be by 50-68 times of concentration in microbial fermentation solution, cell yield is up to 91%.
The object of the present invention is achieved like this, a kind of microbial fermentation solution rapid sedimentation method, comprising the following steps:
Microbial fermentation solution is fitted into container by S1；
Dibasic alkaliine and calcium salt mixed liquor are added into container by S2, and the mixing quality of dibasic alkaliine and calcium salt accounts for
The 5%~10% of quality of fermentation broth；
S3 shakes up above-mentioned solution or is stirred evenly with sterile glass rod, after standing, is placed in 12 in 4 DEG C of environment
~24 hours, supernatant is sopped up, collects the thallus that precipitating is concentration.
In technical solution, dibasic alkaliine and calcium salt are respectively K in S22HPO4And CaCl2, mass ratio be 3:(1.8~
2)；It can also be respectively Na2HPO4And CaCl2, mass ratio is 3:(2~2.2).First quickly 12~15 points of stirring is stirred in S3
Clock is then gently stirred 35~40 minutes, is carrying out slow reaction after fast reaction, so that reaction is more abundant.
Preferably, container is hydrostatic column in S1, specifically can be wide-mouth bottle.Supernatant is sopped up with siphon pipe in S3.
In the present invention, dibasic alkaliine is preferably K2HPO4Or Na2HPO4, calcium salt CaCl2；And calcium salt is first added, it is
Be possible in Nutrient medium containing dibasic alkaliine.In addition, dibasic alkaliine and calcium salt are nontoxic, and low in cost；Phosphorus
Precipitation reaction can occur for sour monohydric salt and calcium salt, generate CaHPO4Precipitating, the deposit are a kind of reticular structure, can be with bacterium
Body cell is bonded together, and accelerates the precipitating of somatic cells, plays the role of accelerating concentration.
By adopting the above-described technical solution, the present invention has the advantage that: it is non-toxic efficient, low in cost, it can will be micro-
50-68 times of concentration in bio-fermented liquid, cell yield is up to 90%.
Specific embodiment
Below with reference to embodiment, the invention will be further described:
Embodiment 1
Bacillus subtilis bacterium solution after taking 1mL shaking table culture 24 hours measures bacterium solution viable bacteria using method of dilution butteron on plate
Number, 500mL bacillus subtilis bacterium solution is transferred in the wide-mouth bottle of 1L, successively adds the CaCl of 3% (m/V)2And K2HPO4
Mixed liquor into bacterium solution, shake up or stirred evenly with sterile glass rod, be put into after static in 4 DEG C of refrigerators, after 12 hours, take
Supernatant is sucked out with siphon pipe after out, collecting precipitating is concentration thallus.
Take 1mL precipitated liquid as follows using method of dilution butteron on plate measurement viable count result with liquid-transfering gun:
Embodiment 2
Bacillus subtilis bacterium solution after taking 1mL shaking table culture 24 hours measures bacterium solution viable bacteria using method of dilution butteron on plate
Number, 500mL bacillus subtilis bacterium solution is transferred in the wide-mouth bottle of 1L, successively adds the CaCl of 5% (m/V)2And K2HPO4
Mixed liquor into bacterium solution, shake up or stirred evenly with sterile glass rod, be put into after static in 4 DEG C of refrigerators, after 12 hours, take
Supernatant is sucked out with siphon pipe after out, collecting precipitating is concentration thallus.
Take 1mL precipitated liquid as follows using method of dilution butteron on plate measurement viable count result with liquid-transfering gun:
Embodiment 3
Bacillus subtilis bacterium solution after taking 1mL shaking table culture 24 hours measures bacterium solution viable bacteria using method of dilution butteron on plate
Number, 500mL bacillus subtilis bacterium solution is transferred in the wide-mouth bottle of 1L, successively adds the CaCl of 8% (m/V)2And K2HPO4
Mixed liquor into bacterium solution, shake up or stirred evenly with sterile glass rod, be put into after static in 4 DEG C of refrigerators, after 12 hours, take
Supernatant is sucked out with siphon pipe after out, collecting precipitating is concentration thallus.
Take 1mL precipitated liquid as follows using method of dilution butteron on plate measurement viable count result with liquid-transfering gun:
Embodiment 4
Bacillus subtilis bacterium solution after taking 1mL shaking table culture 24 hours measures bacterium solution viable bacteria using method of dilution butteron on plate
Number, 500mL bacillus subtilis bacterium solution is transferred in the wide-mouth bottle of 1L, successively adds the CaCl of 10% (m/V)2And K2HPO4
Mixed liquor into bacterium solution, shake up or stirred evenly with sterile glass rod, be put into after static in 4 DEG C of refrigerators, after 12 hours, take
Supernatant is sucked out with siphon pipe after out, collecting precipitating is concentration thallus.
Take 1mL precipitated liquid as follows using method of dilution butteron on plate measurement viable count result with liquid-transfering gun:
Embodiment 5
Bacillus subtilis bacterium solution after taking 1mL shaking table culture 24 hours measures bacterium solution viable bacteria using method of dilution butteron on plate
Number, 500mL bacillus subtilis bacterium solution is transferred in the wide-mouth bottle of 1L, successively adds the CaCl of 12% (m/V)2And K2HPO4
Mixed liquor into bacterium solution, shake up or stirred evenly with sterile glass rod, be put into after static in 4 DEG C of refrigerators, after 12 hours, take
Supernatant is sucked out with siphon pipe after out, collecting precipitating is concentration thallus.
Take 1mL precipitated liquid as follows using method of dilution butteron on plate measurement viable count result with liquid-transfering gun:
1mL precipitated liquid is taken to measure viable count using method of dilution butteron on plate with liquid-transfering gun into embodiment 5 by embodiment 1, it is real
It applies in example 1, the rate of recovery and cycles of concentration are lower, and rate of recovery growth is unobvious in embodiment 5, and cycles of concentration growth is unknown
It is aobvious, thus the mixing quality of dibasic alkaliine and calcium salt accounts for 5%~10% additive amount of quality of fermentation broth, it can be by microorganism
50-68 times of concentration in fermentation liquid, the rate of recovery, that is, cell yield is up to 90%.
In above-described embodiment, dibasic alkaliine and calcium salt are respectively K in S22HPO4And CaCl2, mass ratio be 3:(1.8~
2)；It can also be respectively Na2HPO4And CaCl2, mass ratio is 3:(2~2.2).First quickly 12~15 points of stirring is stirred in S3
Clock is then gently stirred 35~40 minutes, is carrying out slow reaction after fast reaction, so that reaction is more abundant.
Preferably, container is hydrostatic column in S1, specifically can be wide-mouth bottle.Supernatant is sopped up with siphon pipe in S3.
It takes 1mL supernatant to measure viable count using method of dilution butteron on plate with liquid-transfering gun into embodiment 5 embodiment 1, has
The result is as follows:
As seen from the above table, 10%~25% precipitating reagent additive amount is acted on supernatant by significantly reduction, explanation
The precipitating reagent can be preferably by bacterial sediment, and 25% additive amount not significantly reducing compared with 20% illustrates 10%~20%
Precipitating reagent it is more significant to the sedimentation effect of fermentation liquid.Overall cost considers, preferably uses 10%~20% precipitating reagent dosage.
Claims (7)
1. a kind of microbial fermentation solution rapid sedimentation method, which comprises the following steps:
Microbial fermentation solution is fitted into container by S1；
Calcium salt and dibasic alkaliine mixed liquor are successively added into container by S2, and the mixing quality of dibasic alkaliine and calcium salt accounts for
The 5%~10% of quality of fermentation broth；
S3 shakes up above-mentioned solution or is stirred evenly with sterile glass rod, after standing, is placed in 12~24 in 4 DEG C of environment
Hour, supernatant is sopped up, the thallus that precipitating is concentration is collected.
2. a kind of microbial fermentation solution rapid sedimentation method according to claim 1, which is characterized in that phosphoric acid one in S2
Hydrogen salt and calcium salt are respectively K2HPO4And CaCl2, mass ratio is 3:(1.8~2).
3. a kind of microbial fermentation solution rapid sedimentation method according to claim 1, which is characterized in that phosphoric acid one in S2
Hydrogen salt and calcium salt are respectively Na2HPO4And CaCl2, mass ratio is 3:(2~2.2).
4. a kind of microbial fermentation solution rapid sedimentation method according to claim 1, which is characterized in that stirred first in S3
Quickly stirring 12~15 minutes, are then gently stirred 35~40 minutes.
5. a kind of microbial fermentation solution rapid sedimentation method according to claim 1, which is characterized in that container is circle in S1
Cylindrical container.
6. a kind of microbial fermentation solution rapid sedimentation method according to claim 1, which is characterized in that cylinder describes in S1
Device is wide-mouth bottle.
7. a kind of microbial fermentation solution rapid sedimentation method according to claim 1, which is characterized in that use siphon in S3
Pipe sops up supernatant.
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| CN201811530982.XA CN109603212A (en) | 2018-12-14 | 2018-12-14 | A kind of microbial fermentation solution rapid sedimentation method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024046993A1 (en) * | 2022-08-29 | 2024-03-07 | Chr. Hansen A/S | Process for the production of a purified human milk oligosaccharide derived from a microbial fermentation process |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102471755A (en) * | 2009-07-09 | 2012-05-23 | 诺维信公司 | Flocculation with divalent salt and phosphate |
-
2018
- 2018-12-14 CN CN201811530982.XA patent/CN109603212A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102471755A (en) * | 2009-07-09 | 2012-05-23 | 诺维信公司 | Flocculation with divalent salt and phosphate |
Non-Patent Citations (1)
| Title |
|---|
| 杨玉: "《海悦千流：山东大学威海分校本科生科研成果汇编》", 30 April 2012, 山东大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024046993A1 (en) * | 2022-08-29 | 2024-03-07 | Chr. Hansen A/S | Process for the production of a purified human milk oligosaccharide derived from a microbial fermentation process |
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Application publication date: 20190412 |