CN1431036A - Biosurfactant of rhamnolipid and its application in artificial manure turned from house hold garbage - Google Patents
Biosurfactant of rhamnolipid and its application in artificial manure turned from house hold garbage Download PDFInfo
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- CN1431036A CN1431036A CN03118042A CN03118042A CN1431036A CN 1431036 A CN1431036 A CN 1431036A CN 03118042 A CN03118042 A CN 03118042A CN 03118042 A CN03118042 A CN 03118042A CN 1431036 A CN1431036 A CN 1431036A
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- rhamnolipid
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- biological surface
- surface activator
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
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Abstract
A rhamnolipid used as biologic surfactant is prepared from aerugino-pseudomonads through culturing by using plant oil as carbon source and fermenting to obtain fermented solution of rhamnolipid. After centrifugal separation of bacteria and proportional dilution, it can be directly used for preparing compost from lift garbage to improve efficiency and quality of compost.
Description
Technical field: the present invention relates to a kind of tensio-active agent, be specifically related to the bio-surfactant and the application of microorganisms.
Background technology: the improvement of composting process medium physicochemical condition can be by the C/N value, humidity, oxygen, temperature, porosity etc. of control compositing system.The application of these means in composting process is necessary, but the purport of these methods all is to improve mass transfer or favourable microorganism growth, and in the interaction of improvement microorganism and medium and directly promote microorganism aspect the decomposition course of hardly degraded organic substance, to have little scope for one's talents.At present domestic have certain research to biological overall situation improvement aspect in the composting process, and patented technology relates to and more designing and aspect such as improvement for novel appts.Only being then that in the research aspect the microenvironment improving technology physics method of medium improves (as using flyash), does not then appear in the newspapers as yet in the research and development aspect that is used for the bio-surfactant of compost.
There is very big shortcoming in traditional chemical surfactant: because it cannot be degraded by microorganisms, therefore, may bring new pollution to environment, might suppress microbic activity in the compost simultaneously.And bio-surfactant to be vital movement by microorganism form, it does not have toxic action to microorganism, not have the danger of being degraded too early by microorganism as preferential degraded object simultaneously yet, therefore, can be used for the improvement of composting process microenvironment.
Bio-surfactant is a kind of biomolecules by microorganisms, can significantly reduce surface and interface tension force, toxicity to the ecosystem is lower, finally can be degraded by microorganisms, make bio-surfactant can replace the tensio-active agent of chemosynthesis in some respects gradually, and use also more and more widely.Bio-surfactant as microorganisms can strengthen the oils extraction, Daqing Petroleum Administration has developed a kind of rhamnolipid biological surface activator, with other surfactant compound, be used for tertiary oil recovery, average recovery ratio improves 20% (number of patent application 99107581.1) than water.For by the sea of oil pollution or soil, can accelerate the degraded of oils; To the soil of heavy metal contamination and phreatic reparation etc.Li Zuyi etc. have studied solubilising and the desorption of rhamnolipid to polycyclic aromatic hydrocarbons in the soil.Yet, because rhamnolipid is present in the fermented liquid of microorganism, and content is very low, maximum production as the disclosed rhamnolipid of CN1275429A only is 25g/L, and complicated component in the fermented liquid, the technology of the rhamnolipid that separation and Extraction is purer is comparatively complicated, have in its production cost be about 70～90% to be used in all that middle and lower reaches are extracted and separating technology on, thereby the large-scale application of rhamnolipid just is restricted.
Summary of the invention: the present invention is intended to study a kind of rhamnolipid biological surface activator, improves the content of rhamnolipid, is applied to better in the consumer garbage compost processing.
The present invention is achieved through the following technical solutions the foregoing invention purpose.
Rhamnolipid biological surface activator is to be produced through fermentation culture by Pseudomonas aeruginosa (Pseudomonas aeruginose), and content is 30～40g/L.The substratum of producing the Pseudomonas aeruginosa of rhamnolipid is: vegetables oil 60～80g/L, NaNO
33.5～4.5g/L, KH
2PO
41.5～2.2g/L, Na
2HPO
42H
2O 1.5～2.2g/L, MgSO
47H
2O 0.6～1.1g/L, FeSO
47H
2O 0.005～0.012g/L, pH 6.0～6.8, and the culture condition of fermentor tank is 36～38 ℃ of temperature, and 450～550 rev/mins of mixing speed, air flow are 1.1～1.3vvm.Rhamnolipid biological surface activator is as the compost treatment of domestic refuse, the fermented liquid of fermentation culture through centrifugal remove thalline after, be mixed with 0.1～0.6% rhamnolipid solution, with 1～5% uniform mixing of rubbish weight in the domestic refuse of classification.
Description of drawings:
Rubbish interstitial fluid surface tension variations in Fig. 1 compost composting process;
CO in Fig. 2 compost composting process
2Produce rate variation.
The below is described in further detail the present invention.
Pseudomonas aeruginosa (Pseudomonas aeruginose) is available from Chinese Typical Representative culture collection center (CCTCC), and preserving number is AB93066. Strain transfer is deposited on the beef extract-peptone slant medium, and medium component is: beef extract 2.0g/L, peptone 5.0g/L, NaCl 5.0g/L, agar 20.0g/L, pH7.0.
The seed culture based component is: glucose 18.0～25.0g/L, NaNO3 1.5～2.5g/L，KH
2PO
4 0.5～1.2g/L，
Na
2HPO
4·2 H
2O 0.5～1.2g/L，MgSO
4·7H
2O 0.6～1.1g/L，FeSO
4·7H
2O 0.005～0.012g/L, pH 6.0～6.8. The triangular flask liquid amount is 20%, 36～37 ℃ of cultivation temperature, and shaking speed 180～220rpm, after cultivating 24～36 hours, the inoculum concentration with 6～10% is seeded in the fermentation medium.
Fermentation culture based component: vegetables oil 60～80%, NaNO
33.5～4.5g/L, KH
2PO
41.5～2.2g/L, Na
2HPO
42H
2O 1.5～2.2g/L, MgSO
47H
2O 0.6～1.1g/L, FeSO
47H
2O 0.005～0.012g/L, pH6.0～6.8.The culture condition of fermentor tank is that temperature is 36～38 ℃, and 450～550 rev/mins of mixing speed, air flow are 1.1～1.3vvm.
This bacterial strain has stronger bio-surfactant throughput, simultaneously under the inducing action of hydrophobic nature carbon source vegetables oil, makes in the logarithmic growth later stage and produces the amount maximum of rhamnolipid lag phase, reaches 30～40g/L.
Pseudomonas aeruginosa was through 2～4 days cultivation, and fermented liquid is removed thalline through centrifugal (7000-10000rpm), obtains the rhamnolipid product, and the main active ingredient of this product is single rhamnolipid and two rhamnolipids, and its molecular structure is:
Single rhamnolipid molecule
All the other are neutral fat, polysaccharide for two rhamnolipid molecules, and surface tension is 33～35mN/m.
With the rhamnolipid product that obtains, be mixed with 0.1～0.6% rhamnolipid solution, can use by following dual mode, according to the water ratio difference of rubbish, the amount of application of rhamnolipid solution is 1～5% of a rubbish weight:
1. rhamnolipid can add in mixed stirring device, thus in the process that rubbish mix to stir with the abundant mixing of rubbish, the microenvironment of refuse treatment was promptly improved in the starting stage;
2. in the operational process as need add bio-surfactant, then need cooperate humidity control apparatus.The rhamnolipid solution that in the water source of sprinkling system, adds above-mentioned concentration.Along with replenishing of moisture, bio-surfactant just can enter reactor, thereby improves the existence microenvironment of microorganism.
Because composting process is the coefficient complex process of multiple microorganism, the composition of rubbish is also very complicated, and microbial metabolites in the rhamnolipid fermentation liquor and salt can be utilized by microorganism, can the quality of composting production not had a negative impact.
In case rhamnolipid is introduced into the gap of granulated garbage, the hydrophobic side of rhamnolipid molecule will with the organic waste affinity, make molecule be fixed on the granulated garbage surface; Water-wet side is dissolved in the intrapore water of particle.The not same-action of this amphipathic group makes water form stable liquid film on the granulated garbage surface, and the reaction for microorganism of maximum possible provides surperficial environment.Simultaneously, the solubleness of organism in moisture film can be improved in the hydrophobic side of rhamnolipid molecule, thereby quickens microorganism to organic decomposition; In addition, the adding of rhamnolipid also helps the bioavailability that improves hardly degraded organic substance.Measure by experiment and can find, add in the compost reactor of rhamnolipid,, make CO because microorganism is movable vigorous
2Generation speed greater than common compost reactor.After becoming thoroughly decomposed, compost finished product character is better, steady quality.
Embodiment:
1, the preparation of rhamnolipid biological surface activator
1) Pseudomonas aeruginosa is seeded to the seed culture medium from slant medium, carries out shake-flask culture, the bottled nutrient solution 100ml of the triangle of 500ml, 37 ℃, 200rpm shaking culture 24 hours;
2) 4L that packs in the automatic fermenter of 5L is the fermention medium of carbon source with the vegetables oil; Inoculation Pseudomonas aeruginosa seed culture fluid 100mL, 37 ℃ of temperature, 500 rev/mins of mixing speed, air flow is 1.2vvm, and with HCl and NaOH control pH value 6.5, fermentation time is 76 hours, obtain containing the nutrient solution 3.6L of tensio-active agent, the surface tension of fermented liquid is 34.8mN/m after measured;
3) fermented liquid centrifugal (8000rpm) is removed thalline, obtain rhamnolipid solution, by detecting, rhamnolipid content is 37.5g/L;
4) with 15 times of rhamnolipid solution dilutions, obtain 0.25% rhamnolipid solution, standby.Seed culture medium and fermentation culture based component are as follows:
| Seed culture medium | Fermention medium | |
| Glucose | ?18.0g/L | ?- |
| Vegetables oil | ?- | ?50g/L |
| NaNO 3 | ?2.00g/L | ?4.5g/L |
| KH 2PO 4 | ?1.00g/L | ?2.5g/L |
| Na 2HPO 4.2H 2O | ?1.00g/L | ?2.5g/L |
| MgSO 4.7H 2O | ?1.00g/L | ?1.00g/L |
| FeSO 4.7H 2O | ?0.01g/L | ?0.01g/L |
| pH | ?6.5 | ?6.5 |
2, the application in compost
Gather the domestic refuse 20kg in Changsha living quarters, obtain Compostable 8kg through broken, screening, after measured, the water ratio of rubbish is 55%, C/N=25, organic content 60%.Rubbish is packed in the composting device, and the volume of this device is 12L, but load garbage 9L.When packing rubbish into, spray 0.25% rhamnolipid solution, the sprinkling amount is 2% of a rubbish weight.
Extract the rubbish interstitial fluid in the composting process, measure surface tension, detect CO in the composting process simultaneously
2Generation speed, the surface tension and the CO of the rubbish interstitial fluid in the composting process
2Generation speed see Fig. 1 and Fig. 2.
By comparing with the compositing system that does not spray rhamnolipid solution, can find to spray in the compositing system of rhamnolipid solution, the surface tension of rubbish interstitial fluid is lower, and CO
2Generation speed relatively large, compost organic substance degradation rate is very fast, has shortened the time of compost.
Claims (3)
1. a rhamnolipid biological surface activator is characterized in that this bio-surfactant is to be produced through fermentation culture by Pseudomonas aeruginosa (Pseudomonas aeruginose), and rhamnolipid content is 30～40g/L.
2. rhamnolipid biological surface activator according to claim 1 is characterized in that, the fermention medium of producing the Pseudomonas aeruginosa of rhamnolipid is: vegetables oil 60～80g/L, NaNO
33.5～4.5g/L, KH
2PO
41.5～2.2g/L, Na
2HPO
42H
2O 1.5～2.2g/L, MgSO
47H
2O 0.6～1.1g/L, FeSO
47H
2O 0.005～0.012g/L, pH6.0～6.8, the culture condition of fermentor tank is 36～38 ℃ of temperature, 450～550 rev/mins of mixing speed, air flow are 1.1～1.3vvm.
3. the application of a rhamnolipid biological surface activator as claimed in claim 1, it is characterized in that rhamnolipid biological surface activator is used as the composting of domestic refuse, the fermented liquid of Pseudomonas aeruginosa fermentation culture through centrifugal remove thalline after, be mixed with 0.1～0.6% rhamnolipid solution, with 1～5% uniform mixing of rubbish weight in the domestic refuse of classification.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031180426A CN1207383C (en) | 2003-01-23 | 2003-01-23 | Biosurfactant of rhamnolipid and its application in artificial manure turned from house hold garbage |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031180426A CN1207383C (en) | 2003-01-23 | 2003-01-23 | Biosurfactant of rhamnolipid and its application in artificial manure turned from house hold garbage |
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| Publication Number | Publication Date |
|---|---|
| CN1431036A true CN1431036A (en) | 2003-07-23 |
| CN1207383C CN1207383C (en) | 2005-06-22 |
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|---|---|---|---|---|
| CN1326995C (en) * | 2005-01-31 | 2007-07-18 | 湖南大学 | Method for raising green trichoderma cellulase active by biological surfae active agent |
| CN102557772A (en) * | 2012-03-12 | 2012-07-11 | 湖南大学 | Fermentation device for sludge compost treatment and treatment method using same |
| CN103232957A (en) * | 2013-04-18 | 2013-08-07 | 中国科学院微生物研究所 | High-efficiency antibacterial rhamnolipid high-yield strain and application thereof |
| CN103305447A (en) * | 2013-07-04 | 2013-09-18 | 江苏德鑫环保科技有限公司 | Organic waste degradation bacterium and preparation method thereof |
| CN103848701A (en) * | 2014-03-31 | 2014-06-11 | 北京林业大学 | Preparation method of soilless plant cultivation substrate and substrate prepared by method |
| CN104887539A (en) * | 2015-06-24 | 2015-09-09 | 陈竞 | Rhamnolipid-containing personal cleaning product and preparing method thereof |
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| CN1326995C (en) * | 2005-01-31 | 2007-07-18 | 湖南大学 | Method for raising green trichoderma cellulase active by biological surfae active agent |
| CN102557772A (en) * | 2012-03-12 | 2012-07-11 | 湖南大学 | Fermentation device for sludge compost treatment and treatment method using same |
| CN103232957A (en) * | 2013-04-18 | 2013-08-07 | 中国科学院微生物研究所 | High-efficiency antibacterial rhamnolipid high-yield strain and application thereof |
| CN103232957B (en) * | 2013-04-18 | 2015-04-08 | 中国科学院微生物研究所 | High-efficiency antibacterial rhamnolipid high-yield strain and application thereof |
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| CN103848701A (en) * | 2014-03-31 | 2014-06-11 | 北京林业大学 | Preparation method of soilless plant cultivation substrate and substrate prepared by method |
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| CN106495871A (en) * | 2016-11-01 | 2017-03-15 | 西北农林科技大学 | A kind of method of macrolide resistant gene and intI1 abundance in reduction Chicken Manure Compost |
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