CN109609411A - A kind of microbial flora and preparation method thereof for erythromycin of degrading - Google Patents
A kind of microbial flora and preparation method thereof for erythromycin of degrading Download PDFInfo
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- CN109609411A CN109609411A CN201910016342.5A CN201910016342A CN109609411A CN 109609411 A CN109609411 A CN 109609411A CN 201910016342 A CN201910016342 A CN 201910016342A CN 109609411 A CN109609411 A CN 109609411A
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- erythromycin
- degrading
- composite bacteria
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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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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
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- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
Abstract
The microbial composite bacteria group and preparation method thereof that the present invention provides a kind of for erythromycin of degrading, belongs to biodegradable field.The microbial composite bacteria group is made of Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae, each component volume ratio is 1-3:1-3:1-3, and Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae are cultivated 3-4 days total bacteria count 10 into bacterium solution under the conditions of on beef extract-peptone fluid nutrient medium in 35 DEG C by prescription amount8A/mL；Liquid spawn is transferred in the minimal medium containing erythromycin, the amount ratio of the liquid spawn and minimal medium is 2:98 (w/w), 3 days is cultivated under the conditions of 35 DEG C to obtain the final product.Operation of the present invention is easy, is remarkably improved the degradation rate of erythromycin, preferably protects environment.
Description
Technical field
It is especially a kind of for the micro- of erythromycin of degrading the present invention relates to a kind of microbial composite bacteria group and preparation method thereof
Biological compound fungi group and preparation method thereof.
Background technique
As novel micro pollutant drug and personal-care supplies (Pharmaceutical and personal care
One of products, PPCPs), erythromycin is determined as emerging pollutant by United Nations Environment Programme, excessively uses meeting
Human body is caused to seriously endanger, meeting liver injury system, renal system, hemopoietic system etc..It is useless that erythromycin is primarily present in medicine
In object and sewage, soil can also be polluted in conjunction with biosolids, and then influence environment and pass through sewage in the environment
It propagates, ecology is impacted.It is detected in water environment mostly in recent years as pharmaceutical production and consumption big country in China
Erythromycin out.Therefore, it is necessary to carry out degradation treatment to erythromycin since developing one kind more method of novel environment friendly.
The Study on degradation of erythromycin focuses primarily upon physical method and chemical method, but at high cost needed for the two, experiment
Condition is stringent, and the substance generated therewith is also easy to carry out secondary pollution to environment.Biodegrade refers to using microorganism to red mould
Element is degraded, and for physics and chemical method, can be reduced required cost, be reduced the pollution to environment.Therefore it builds
A kind of microbial composite bacteria group for erythromycin of degrading is stood, is had great importance.
Summary of the invention
Technical problem to be solved by the present invention lies in provide a kind of microbial composite bacteria group for erythromycin of degrading.
Another technical problem to be solved by this invention is to provide the above-mentioned microbial composite bacteria for erythromycin of degrading
The preparation method of group.
In order to solve the above technical problems, the technical scheme is that
A kind of microbial composite bacteria group for erythromycin of degrading, by Bacillus cereus, pseudomonas aeruginosa and pneumonia
Klebsiella composition, each component volume ratio are 1-3:1-3:1-3.
The preferred above-mentioned microbial composite bacteria group for erythromycin of degrading, the Bacillus cereus, P. aeruginosa
The volume ratio of bacterium and Klebsiella Pneumoniae is 1:1:3.
The preparation method of the above-mentioned microbial composite bacteria group for erythromycin of degrading, specific preparation process is as follows:
(1) prepared by liquid spawn: by prescription amount by Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae in ox
It is cultivated 3-4 days under the conditions of 35 DEG C on meat extract peptone fluid nutrient medium；
(2) liquid spawn is transferred in the minimal medium containing erythromycin, the liquid spawn and inorganic salts culture
The amount ratio of base is 2:98 (w/w), is cultivated 3 days under the conditions of 35 DEG C.The amount ratio of the minimal medium is Erythromycin A
0.01g/L、(NH4)2SO41.2-1.7g/L, peptone 0.4-0.8g/L, KH2PO4 0.3-0.7g/L、K2HPO4 1.1-
1.6g/L、MgSO4·7H2O 0.1-0.3g/L、NaCl 0.3-0.5g/L。
The preparation method of the preferred above-mentioned microbial composite bacteria group for erythromycin of degrading, it is inorganic in the step (2)
Salt culture medium is matched by content: Erythromycin A 0.01g/L, (NH4)2SO41.5g/L, peptone 0.5g/L, KH2PO4 0.5g/L、
K2HPO4 1.5g/L、MgSO4·7H2O0.2g/L, NaCl 0.5g/L are obtained.
Beneficial effects of the present invention:
The above-mentioned microbial composite bacteria group for erythromycin of degrading, has good synergistic effect, can significantly improve red
The degradation rate of mycin, has certain application value, and application prospect is extensive.
Specific embodiment
Technical solution of the present invention is further described combined with specific embodiments below.
Embodiment 1
A kind of microbial composite bacteria group for erythromycin of degrading, by Bacillus cereus, pseudomonas aeruginosa and pneumonia
Klebsiella composition, each component volume ratio are 1:3:3.
It is above-mentioned for degrade erythromycin microbial composite bacteria group's the preparation method is as follows:
(1) prepared by liquid spawn: by prescription amount by Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae in ox
It is cultivated 3 days under the conditions of 35 DEG C on meat extract peptone fluid nutrient medium, until total bacteria count 10 in bacterium solution8A/mL；
(2) liquid spawn is transferred in the minimal medium containing erythromycin, the liquid spawn and inorganic salts culture
The amount ratio of base is 2:98 (w/w), is cultivated 3 days under the conditions of 35 DEG C, and the minimal medium is matched by content: Erythromycin A
0.01g/L、(NH4)2SO41.4g/L, peptone 0.5g/L, KH2PO4 0.5g/L、K2HPO4 1.5g/L、MgSO4·7H2O
0.2g/L, NaCl 0.5g/L are obtained.
The BPDA fluid nutrient medium: taking beef extract 5.0g, peptone 10.0g, NaCl 5.0g, and deionized water is added
1000mL is all put into autoclave after dissolution, high temperature (120 DEG C) high pressure steam sterilization 20 minutes.
Embodiment 2
A kind of microbial composite bacteria group for erythromycin of degrading, by Bacillus cereus, pseudomonas aeruginosa and pneumonia
Klebsiella composition, each component volume ratio are 2:1:2.
It is above-mentioned for degrade erythromycin microbial composite bacteria group's the preparation method is as follows:
(1) prepared by liquid spawn: by prescription amount by Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae in ox
It is cultivated 4 days under the conditions of 35 DEG C on meat extract peptone fluid nutrient medium, until total bacteria count 10 in bacterium solution8A/mL；
(2) liquid spawn is transferred in the minimal medium containing erythromycin, the liquid spawn and inorganic salts culture
The amount ratio of base is 2:98 (w/w), is cultivated 3 days under the conditions of 35 DEG C, and the minimal medium is matched by content: Erythromycin A
0.01g/L、(NH4)2SO41.5g/L, peptone 0.8g/L, KH2PO4 0.3g/L、K2HPO4 1.5g/L、MgSO4·7H2O
0.2g/L, NaCl 0.4g/L are obtained.
Embodiment 3
A kind of microbial composite bacteria group for erythromycin of degrading, by Bacillus cereus, pseudomonas aeruginosa and pneumonia
Klebsiella composition, each component volume ratio are 2:2:3.
It is above-mentioned for degrade erythromycin microbial composite bacteria group's the preparation method is as follows:
(1) prepared by liquid spawn: by prescription amount by Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae in ox
It is cultivated 3 days under the conditions of 35 DEG C on meat extract peptone fluid nutrient medium, until total bacteria count 10 in bacterium solution8A/mL；
(2) liquid spawn is transferred in the minimal medium containing erythromycin, the liquid spawn and inorganic salts culture
The amount ratio of base is 2:98 (w/w), is cultivated 3 days under the conditions of 35 DEG C, and the minimal medium is matched by content: erythromycin
A0.01g/L、(NH4)2SO41.6g/L, peptone 0.5g/L, KH2PO4 0.7g/L、K2HPO4 1.5g/L、MgSO4·7H2O
0.1g/L, NaCl 0.5g/L are obtained.
Embodiment 4
A kind of microbial composite bacteria group for erythromycin of degrading, by Bacillus cereus, pseudomonas aeruginosa and pneumonia
Klebsiella composition, each component volume ratio are 3:2:1.
It is above-mentioned for degrade erythromycin microbial composite bacteria group's the preparation method is as follows:
(1) prepared by liquid spawn: by prescription amount by Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae in ox
It is cultivated 3 days under the conditions of 35 DEG C on meat extract peptone fluid nutrient medium, until total bacteria count 10 in bacterium solution8A/mL；
(2) liquid spawn is transferred in the minimal medium containing erythromycin, the liquid spawn and inorganic salts culture
The amount ratio of base is 2:98 (w/w), is cultivated 3 days under the conditions of 35 DEG C, and the minimal medium is matched by content: erythromycin
A0.01g/L、(NH4)2SO41.2g/L, peptone 0.4g/L, KH2PO4 0.3g/L、K2HPO4 1.3g/L、MgSO4·7H2O
0.2g/L, NaCl 0.4g/L are obtained.
Embodiment 5
A kind of microbial composite bacteria group for erythromycin of degrading, by Bacillus cereus, pseudomonas aeruginosa and pneumonia
Klebsiella composition, each component volume ratio are 3:1:3.
It is above-mentioned for degrade erythromycin microbial composite bacteria group's the preparation method is as follows:
(1) prepared by liquid spawn: by prescription amount by Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae in ox
It is cultivated 4 days under the conditions of 35 DEG C on meat extract peptone fluid nutrient medium, until total bacteria count 10 in bacterium solution8A/mL；
(2) liquid spawn is transferred in the minimal medium containing erythromycin, the liquid spawn and inorganic salts culture
The amount ratio of base is 2:98 (w/w), is cultivated 3 days under the conditions of 35 DEG C, and the minimal medium is matched by content: erythromycin
A0.01g/L、(NH4)2SO41.5g/L, peptone 0.5g/L, KH2PO4 0.5g/L、K2HPO4 1.5g/L、MgSO4·7H2O
0.2g/L, NaCl 0.5g/L are obtained.
Test example
Degrading experiment is carried out to erythromycin using microbial composite bacteria group described in embodiment 1-5, is tested as follows:
Experimental material: erythromycin solution, microbial flora
Test method: being dissolved to concentration 100mg/L with methanol for erythromycin powder, and made flora is added by throwing bacterium amount, puts
Enter shaking table concussion, each processing is arranged 3 in parallel, carries out high performance liquid chromatography in third day and measures erythromycin surplus, calculates
Erythromycin degradation rate.Experimental setup 6 processing, processing 1 add the made microbial flora of experimental example 1 by throwing bacterium amount 4%；Processing
2 add the made microbial flora of experimental example 2 by throwing bacterium amount 4%；Processing 3 adds the made micro- life of experimental example 3 by throwing bacterium amount 4%
Object flora；Processing 4 adds the made microbial flora of experimental example 4 by throwing bacterium amount 4%, and processing 5 adds experimental example by throwing bacterium amount 4%
5 made microbial floras, processing 6 are not added with flora.
Experimental result: as shown in table 1, the erythromycin degradation rate for handling 1-5 is above blank control, 3 days degradation rates point
Not are as follows: processing 1 is 91.47%, and processing 2 is 90.20%, and processing 3 is 93.67%, and processing 4 is 88.34%, and processing 5 is
95.28%, processing 6 is 5.16%.
Conclusion: degrading microorganism flora can promote the degradation of erythromycin, improve degradation rate.
Table 1
| Processing | Erythromycin degradation rate % |
| Processing 1 | 91.47 |
| Processing 2 | 90.20 |
| Processing 3 | 93.67 |
| Processing 4 | 88.34 |
| Processing 5 | 95.28 |
| Processing 6 | 5.16 |
What above-mentioned microbial composite bacteria group referring to embodiment to this kind for erythromycin of degrading and preparation method thereof carried out
It is discussed in detail, is illustrative without being restrictive, several embodiments can be enumerated according to limited range, therefore not
The change and modification being detached under present general inventive concept, should belong within protection scope of the present invention.
Claims (4)
1. a kind of microbial composite bacteria group for erythromycin of degrading, it is characterised in that: by Bacillus cereus, P. aeruginosa
Bacterium and Klebsiella Pneumoniae composition, each component volume ratio are 1-3:1-3:1-3.
2. the microbial composite bacteria group according to claim 1 for erythromycin of degrading, it is characterised in that: the waxy bud
The volume ratio of born of the same parents bacillus, pseudomonas aeruginosa and Klebsiella Pneumoniae is 1:1:3.
3. the preparation method described in claim 1 for the microbial composite bacteria group for erythromycin of degrading, it is characterised in that: specific system
It is standby that steps are as follows:
(1) prepared by liquid spawn: by prescription amount by Bacillus cereus, pseudomonas aeruginosa and Klebsiella Pneumoniae in beef extract
It is cultivated 3-4 days under the conditions of 35 DEG C on peptone fluid nutrient medium, until total bacteria count 10 in bacterium solution8A/mL；
(2) liquid spawn is transferred in the minimal medium containing erythromycin, the liquid spawn and minimal medium
Amount ratio is 2:98 (w/w), is cultivated 3 days under the conditions of 35 DEG C；The amount ratio of the minimal medium is Erythromycin A
0.01g/L、(NH4)2SO41.2-1.7g/L, peptone 0.4-0.8g/L, KH2PO4 0.3-0.7g/L、K2HPO4 1.1-
1.6g/L、MgSO4·7H2O 0.1-0.3g/L、NaCl 0.3-0.5g/L。
4. the preparation method of the microbial composite bacteria group according to claim 3 for erythromycin of degrading, it is characterised in that:
Minimal medium is matched by content in the step (2): Erythromycin A 0.01g/L, (NH4)2SO41.5g/L, albumen
Peptone 0.5g/L, KH2PO4 0.5g/L、K2HPO4 1.5g/L、MgSO4·7H2O 0.2g/L、NaCl 0.5g/L。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111893052A (en) * | 2020-05-25 | 2020-11-06 | 常州大学 | Erythromycin-degrading bacterium pseudomonas aeruginosa RJJ-1 and application thereof |
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| CN103540549A (en) * | 2013-10-24 | 2014-01-29 | 哈尔滨工业大学 | Preparation method of compound microbial inoculum for degrading domestic sewage |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111893052A (en) * | 2020-05-25 | 2020-11-06 | 常州大学 | Erythromycin-degrading bacterium pseudomonas aeruginosa RJJ-1 and application thereof |
| CN111893052B (en) * | 2020-05-25 | 2022-06-24 | 常州大学 | Erythromycin-degrading bacterium pseudomonas aeruginosa RJJ-1 and application thereof |
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