CN107663511A - A kind of culture medium of nitrite bacteria and preparation method thereof - Google Patents
A kind of culture medium of nitrite bacteria and preparation method thereof Download PDFInfo
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- CN107663511A CN107663511A CN201711237611.8A CN201711237611A CN107663511A CN 107663511 A CN107663511 A CN 107663511A CN 201711237611 A CN201711237611 A CN 201711237611A CN 107663511 A CN107663511 A CN 107663511A
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- 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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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
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Abstract
The present invention relates to cell culture medium technical field, more particularly to a kind of culture medium for cultivating nitrite bacteria and preparation method thereof, the culture medium of the culture nitrite bacteria, including basal medium and additive, it is characterised in that the additive includes following material：Solid absorption thing, metal salt, ammonium salt, organic amine, pH buffer, stimulating factor；It can improve the bacteria concentration of nitrobacteria by the medium culture nitrite bacteria of the present invention, bacteria concentration is up to 10 after batch cultivation9Individual/mL, the bacterial concentration that more existing cultural method obtains improve more than 10 times.
Description
Technical field
The present invention relates to cell culture medium technical field, more particularly to a kind of culture medium and its system for cultivating nitrite bacteria
Preparation Method.
Background technology
At present, increasingly paid attention to the problem of ammonia nitrogen pollution in water body, aquaculture, eutrophic lake are administered, high ammonia nitrogen
All there is an urgent need to carry out the removal of ammonia nitrogen for many application fields such as Industrial Wastewater Treatment.The removal of water body ammonia nitrogen exist physico-chemical process and
The major class of bioanalysis two.Physico-chemical process such as ammonia-nitrogen desorption method, the method needs to use a large amount of highly basic, and final ammonia nitrogen is discharged into air,
Not only processing cost is higher, secondary pollution be present, and is only capable of being directed to high-concentration ammonia nitrogenous wastewater, and bioanalysis, in particular with thin
Bacterium except ammonia method due to it to well adapting to property of ammonia nitrogen, processing cost is low, non-secondary pollution the features such as and application more
Extensively, it is more concerned.
Synchronous nitration and denitrification（SND）This Process of Biological Nitrogen Removal of denitrogenation can be carried out simultaneously in same reactor
Ammonia nitrogen removal and denitrification removing total nitrogen are nitrified, not only overcomes some problems existing for traditional biological denitrification process, and reducing
Energy consumption and material consumption etc. have prominent advantage：It can effectively keep pH in reactor stable, reduce or cancel the throwing of basicity
Add；Reduce the volume of traditional reactor, save capital cost；For the sequencing batch reactor being only made up of a reaction tank, SND
It can reduce and realize time needed for nitration denitrification；Save aeration quantity, further reduce energy consumption.Since the 1980s,
The research of bioscience man finds many nitrifier such as Pseudomonas fluorescens（Pseudomonas flurescens）, excrement production bacillus
（Alcaligenes facealis）, pseudomonas aeruginosa (Pseudomonas aeruginos) etc. can be to organic or nothing
Machine ammoniate carries out heterotrophic nitrification.Compared with autotrophic type nitrifier, although the speed of the heterotroph oxidation ammonium salt of biomass
The rate 2-3 order of magnitude slower than autotrophic bacterium, but the speed of growth of allotrophic nitrobacteria is fast, cell yield is high；It is required that dissolved oxygen is dense
Spend low；It is also strong to the adaptability of environment, more sour environment can be endured, the speed of its overall oxidation ammonium salt is unlike autotrophic bacterium
Slowly.
Nitrite bacteria is distributed very generally in nature, nitrite bacteria all be present in soil, fresh water and ocean, he
One of two physiology subgroups of Nitrobacteriaceae can be belonged to by ammoxidation into nitrous acid, in the nitrification first step, i.e., by ammonium salt
Worked during being oxidized to nitrite, while be also the relation of its first rate-limiting reaction, nitrite bacteria and the mankind
Very closely, it is widely used in food, biology, medical industry purification of waste water, municipal sewage treatment, fish reproduction, crops
In terms of soil improvement.Because nitrobacteria is chemoautotrophic bacteria, it has using ammonia nitrogen as unique energy source material, strict aerobic, life
Long speed is slow, the average physiological property for duration, causes that its industrialized production cycle is long, aeration energy consumption is big, production cost is high, sternly
Its popularization and application in terms of ammonia nitrogen pollution in water body improvement is limited again.
The content of the invention
In view of the shortcomings of the prior art, it is an object of the invention to provide a kind of culture medium of nitrite bacteria and its preparation
Method, the culture medium of nitrite bacteria provided by the invention, good condition is provided for the growth and breeding of nitrite bacteria, shortened
Its production cycle, reduce production cost.
To solve the above problems, the invention provides a kind of nitrite bacteria culture medium, including basal medium and addition
Agent, the additive include following material：Solid absorption thing, metal salt, ammonium salt, organic amine, pH buffer, stimulating factor.
The present invention does not have special requirement to the species of basal medium, can often know for those skilled in the art, example
Such as, the basal medium is F12 or the culture mediums of RMPI 1640.
The content of each material is to influence the most important factor of nitrite bacteria growth and breeding in culture medium, it is therefore desirable to adding
The content of each material in agent is added to carry out Reasonable adjustment, it is preferred that the additive includes the material of following parts by weight：Solid absorption
3 ~ 20 parts of thing, 15 ~ 50 parts of metal salt, 5 ~ 25 parts of ammonium salt, 0.5 ~ 3 part of organic amine, 0.1 ~ 20 part of pH buffer, stimulating factor 0.1 ~
1 part.
Solid absorption thing can provide place for the growth and breeding of nitrite bacteria, while be also easy to separation of bacterial, this hair
Solid absorption thing in bright is that density is less than water, and the material that can be swum in water can be activated carbon, porous carbon, stalk, wood
At least one of material, preferably stalk, timber.
Preferably, the metal salt can provide nutrition for the growth of nitrite bacteria, be to influence to nitrify thin growth and breeding
One of an important factor for, the preferable metal salt is sodium salt, sylvite, molysite, calcium salt, manganese salt, nickel salt, mantoquita, magnesium salts, ferrous iron
At least one of salt.
The sodium salt can be at least one in sodium chloride, sodium sulphate, sodium nitrate, sodium dihydrogen phosphate, disodium hydrogen phosphate etc.
Kind.The sylvite can be at least one of potassium chloride, potassium sulfate, potassium nitrate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate etc..Institute
It can be at least one of iron chloride, frerrous chloride, ferric sulfate, ferrous sulfate to state molysite.The calcium salt is calcium chloride, sulfuric acid
At least one of calcium.The mantoquita is at least one of copper chloride, copper sulphate.The magnesium salts is in magnesium chloride, magnesium sulfate
At least one.The nickel salt is at least one of nickel chloride, nickel sulfate.The manganese salt is manganese chloride, in manganese sulfate extremely
Few one kind.
The concentration of metal salt is to influence the most important factor of nitrite bacteria growth and breeding, and concentration is too low, can not provide and fill
The nutrition of foot, excessive concentration, then can cause bacterium metal poisoning, suppress the growth of bacterium.Preferably, the concentration of the metal salt
For 1 ~ 30g/L.
Preferably, the ammonium salt is at least one of ammonium chloride, ammonium nitrate, ammonium sulfate, ammonium nilrite, ammonium carbonate.
Preferably, the organic amine is at least one of amino acid, spermine, spermidine, formic acid azanol, hydroxylamine acetate.
It is anti-that the addition of organic acid azanol can directly participate in the metabolic process of nitrobacteria, shortening enzymatic as the matrix of azanol oxygen also enzyme
Answer process, accelerated cell growth.Preferably organic acid azanol, the organic acid that organic acid azanol discharges also help some heterotrophism
The growth of nitrobacteria, be advantageous to play the population effect of nitrobacteria, be favorably improved the ability of flora tolerance low temperature environment.
Stimulating factor can stimulate nitrobacteria to be grown, and reduce the production time, it is preferred that the stimulating factor is lanthanum
It is at least one of metal salt.
A kind of method for cultivating nitrite bacteria, comprises the following steps：
（1）Metal salt, ammonium salt, organic amine, stimulating factor are added in basal medium, after being well mixed, obtains mixed system
1；
（2）The pH value for adjusting mixed system is 7.5 ~ 8.0, then adds solid absorption thing, and be inoculated with nitrite bacteria；
（3）The pH value for maintaining culture medium is 7.5 ~ 8.0, under conditions of temperature is 28 ~ 35 DEG C, humidity is 60 ~ 85%, and is continuously exposed
Gas or intermittent aerating, after ammonia nitrogen runs out of in culture medium, filtered with micron membrane filter, after degerming, produce nitrite bacteria.
Preferably, the micron membranes are the filter membrane that aperture is 0.1 ~ 0.3 micron.
Compared with prior art, the present invention has following technique effect：
The culture medium of the present invention can realize the fast breeding of nitrite bacteria, shorten enrichment by scientific and reasonable proportioning
Incubation time.
It can improve the bacteria concentration of nitrobacteria by the medium culture nitrite bacteria of the present invention, after batch cultivation
Bacteria concentration is up to 109Individual/mL, the bacterial concentration that more existing cultural method obtains improve more than 10 times.
The present invention is by adding organic solid absorption thing that suspends, it is possible to achieve intermittent aerating culture, ventilation is reduced, so as to drop
Low production cost.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, following examples, the present invention is carried out
It is further described.It should be appreciated that specific embodiment described herein is not used to limit this only to explain the present invention
Invention.
Embodiment 1
A kind of nitrite bacteria culture medium is present embodiments provided, including basal medium and additive, the additive include
Following material：
10 parts of activated carbon, 30 parts of sodium salt, 18 parts of ammonium chloride, amino acid/11 .5 parts, 10 parts of pH buffer, 0.5 part of lanthanide metal salt；
The present embodiment additionally provides a kind of method for cultivating nitrite bacteria, using above-mentioned medium culture, comprises the following steps：
（1）Sodium salt, ammonium chloride, amino acid, lanthanide metal salt are added in basal medium, after being well mixed, obtains mixture
It is 1；
（2）The pH value for adjusting mixed system is 7.6, then adds activated carbon, and be inoculated with nitrite bacteria；
（3）The pH value for maintaining culture medium is 7.7, and under conditions of temperature is 30 DEG C, humidity is 70%, and continuous aeration or interval expose
Gas, after ammonia nitrogen runs out of in culture medium, with the membrane filtration that aperture is 0.2 micron, after degerming, produce nitrite bacteria.
Embodiment 2
A kind of nitrite bacteria culture medium is present embodiments provided, including basal medium and additive, the additive include
Following material：
4 parts of porous carbon, 16 parts of sylvite, 7 parts of ammonium sulfate, 0.8 part of spermidine, 0.8 part of pH buffer, 0.5 part of lanthanide metal salt；
The present embodiment additionally provides a kind of method for cultivating nitrite bacteria, using above-mentioned medium culture, comprises the following steps：
（1）Sylvite, ammonium sulfate, spermidine, lanthanide metal salt are added in basal medium, after being well mixed, obtains mixture
It is 1；
（2）The pH value for adjusting mixed system is 7.7, then adds porous carbon, and be inoculated with nitrite bacteria；
（3）The pH value for maintaining culture medium is 7.7, and under conditions of temperature is 29 DEG C, humidity is 68%, and continuous aeration or interval expose
Gas, after ammonia nitrogen runs out of in culture medium, with the membrane filtration that aperture is 0.1 micron, after degerming, produce nitrite bacteria.
Embodiment 3
A kind of nitrite bacteria culture medium is present embodiments provided, including basal medium and additive, the additive include
Following material：
18 parts of stalk, 45 parts of molysite, 24 parts of ammonium nilrite, 2.5 parts of formic acid azanol, 18 parts of pH buffer, lanthanide metal salt 0.8
Part；
The present embodiment additionally provides a kind of method for cultivating nitrite bacteria, using above-mentioned medium culture, comprises the following steps：
（1）Molysite, ammonium nilrite, formic acid azanol, lanthanide metal salt are added in basal medium, after being well mixed, is mixed
Zoarium system 1；
（2）The pH value for adjusting mixed system is 8.0, then adds stalk, and be inoculated with nitrite bacteria；
（3）The pH value for maintaining culture medium is 8.0, and under conditions of temperature is 35 DEG C, humidity is 85%, and continuous aeration or interval expose
Gas, after ammonia nitrogen runs out of in culture medium, with the membrane filtration that aperture is 0.3 micron, after degerming, produce nitrite bacteria.
Embodiment 4
A kind of nitrite bacteria culture medium is present embodiments provided, including basal medium and additive, the additive include
Following material：
3 parts of timber, 15 parts of calcium salt, 5 parts of ammonium carbonate, 0.5 part of hydroxylamine acetate, 0.1 part of pH buffer, 0.1 part of lanthanide metal salt；
The present embodiment additionally provides a kind of method for cultivating nitrite bacteria, using above-mentioned medium culture, comprises the following steps：
（1）Calcium salt, ammonium carbonate, hydroxylamine acetate, lanthanide metal salt are added in basal medium, after being well mixed, is mixed
System 1；
（2）The pH value for adjusting mixed system is 7.5, then adds timber, and be inoculated with nitrite bacteria；
（3）The pH value for maintaining culture medium is 7.5, and under conditions of temperature is 28 DEG C, humidity is 60%, and continuous aeration or interval expose
Gas, after ammonia nitrogen runs out of in culture medium, with the membrane filtration that aperture is 0.1 micron, after degerming, produce nitrite bacteria.
Embodiment 5
A kind of nitrite bacteria culture medium is present embodiments provided, including basal medium and additive, the additive include
Following material：
20 parts of activated carbon, 50 parts of manganese salt, 25 parts of ammonium nilrite, 3 parts of spermidine, 20 parts of pH buffer, 1 part of lanthanide metal salt；
The present embodiment additionally provides a kind of method for cultivating nitrite bacteria, using above-mentioned medium culture, comprises the following steps：
（1）Manganese salt, ammonium nilrite, spermidine, lanthanide metal salt are added in basal medium, after being well mixed, is mixed
System 1；
（2）The pH value for adjusting mixed system is 7.5, then adds activated carbon, and be inoculated with nitrite bacteria；
（3）The pH value for maintaining culture medium is 7.5, and under conditions of temperature is 29 DEG C, humidity is 70%, and continuous aeration or interval expose
Gas, after ammonia nitrogen runs out of in culture medium, with the membrane filtration that aperture is 0.2 micron, after degerming, produce nitrite bacteria.
Foregoing description is only the description to section Example of the present invention, not to any restriction of the scope of the invention, one's own profession
The those of ordinary skill of industry can make improvement or modification according to the present invention to above-described embodiment, but belong to present invention protection model
Enclose.
Claims (9)
1. a kind of nitrite bacteria culture medium, including basal medium and additive, it is characterised in that the additive include with
Lower material：Solid absorption thing, metal salt, ammonium salt, organic amine, pH buffer, stimulating factor.
2. nitrite bacteria culture medium according to claim 1, it is characterised in that the additive includes following parts by weight
Material：3 ~ 20 parts of solid absorption thing, 15 ~ 50 parts of metal salt, 5 ~ 25 parts of ammonium salt, 0.5 ~ 3 part of organic amine, pH buffer 0.1 ~ 20
Part, 0.1 ~ 1 part of stimulating factor.
3. nitrite bacteria culture medium according to claim 1, it is characterised in that the solid absorption thing be activated carbon,
At least one of porous carbon, stalk, timber.
4. nitrite bacteria culture medium according to claim 1, it is characterised in that the metal salt is sodium salt, sylvite, iron
At least one of salt, calcium salt, manganese salt, nickel salt, mantoquita, magnesium salts, ferrous salt.
5. nitrite bacteria culture medium according to claim 1, it is characterised in that the ammonium salt be ammonium chloride, ammonium nitrate,
At least one of ammonium sulfate, ammonium nilrite, ammonium carbonate.
6. nitrite bacteria culture medium according to claim 1, it is characterised in that the organic amine be amino acid, spermine,
At least one of spermidine, formic acid azanol, hydroxylamine acetate.
7. nitrite bacteria culture medium according to claim 1, it is characterised in that the stimulating factor is lanthanide metal salt
At least one of.
A kind of 8. method for cultivating nitrite bacteria, it is characterised in that using the culture medium described in claim 1 ~ 7 any one
Culture, comprises the following steps：
（1）Metal salt, ammonium salt, organic amine, stimulating factor are added in basal medium, after being well mixed, obtains mixed system
1；
（2）The pH value for adjusting mixed system is 7.5 ~ 8.0, then adds solid absorption thing, and be inoculated with nitrite bacteria；
（3）The pH value for maintaining culture medium is 7.5 ~ 8.0, under conditions of temperature is 28 ~ 35 DEG C, humidity is 60 ~ 85%, and is continuously exposed
Gas or intermittent aerating, after ammonia nitrogen runs out of in culture medium, filtered with micron membrane filter, after degerming, produce nitrite bacteria.
9. the method for culture nitrite bacteria according to claim 8, it is characterised in that the micron membranes are that aperture is
0.1 ~ 0.3 micron of filter membrane.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114686471A (en) * | 2020-12-30 | 2022-07-01 | 中国石油化工股份有限公司 | Nitrosation microbial inoculum and preparation method thereof |
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| CN104711213A (en) * | 2015-03-13 | 2015-06-17 | 中国海洋石油总公司 | Accelerant facilitating growth of salt-resistive nitrifying bacteria in high-salt wastewater |
| CN105624093A (en) * | 2014-10-28 | 2016-06-01 | 中国石油化工股份有限公司 | Nitrite bacterium culture promoter and preparation method thereof |
| CN105670964A (en) * | 2016-01-27 | 2016-06-15 | 山东省科学院生物研究所 | Bacillus atrophaeus strain BsR05 and application thereof |
-
2017
- 2017-11-30 CN CN201711237611.8A patent/CN107663511A/en not_active Withdrawn
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101899401A (en) * | 2009-05-25 | 2010-12-01 | 中国石油化工股份有限公司 | Microbial agent for treating ammonia-containing waste water and preparation method thereof |
| CN101928689A (en) * | 2010-07-13 | 2010-12-29 | 王栋 | Method for industrially preparing Rhodobacter sphaeroides, fermentation liquor of Rhodobacter sphaeroides and application |
| CN105624093A (en) * | 2014-10-28 | 2016-06-01 | 中国石油化工股份有限公司 | Nitrite bacterium culture promoter and preparation method thereof |
| CN104711213A (en) * | 2015-03-13 | 2015-06-17 | 中国海洋石油总公司 | Accelerant facilitating growth of salt-resistive nitrifying bacteria in high-salt wastewater |
| CN105670964A (en) * | 2016-01-27 | 2016-06-15 | 山东省科学院生物研究所 | Bacillus atrophaeus strain BsR05 and application thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114686471A (en) * | 2020-12-30 | 2022-07-01 | 中国石油化工股份有限公司 | Nitrosation microbial inoculum and preparation method thereof |
| CN114686471B (en) * | 2020-12-30 | 2024-01-05 | 中国石油化工股份有限公司 | Nitrosation microbial inoculum and preparation method thereof |
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Application publication date: 20180206 |