CN104176803A - Azotobacter chroococcum biological flocculant and preparation method thereof - Google Patents
Azotobacter chroococcum biological flocculant and preparation method thereof Download PDFInfo
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- CN104176803A CN104176803A CN201410412875.2A CN201410412875A CN104176803A CN 104176803 A CN104176803 A CN 104176803A CN 201410412875 A CN201410412875 A CN 201410412875A CN 104176803 A CN104176803 A CN 104176803A
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Abstract
The invention relates to an azotobacter chroococcum biological flocculant and a preparation method thereof. Because the preparation of a microbial flocculant is high in culture cost, long in period and needs solid-liquid separation after fermentation, a product is increased in cost. The preparation method comprises the following steps: adding water to mannitol, KH2PO4, MgSO4.7H2O, NaCl, CaCO3, CaSO4.2H2O to prepare a fermentation culture medium; inoculating a solid slant strain of activated azotobacter chroococcum to the fermentation culture medium for culture to obtain a fermentation seed solution, and inoculating the fermentation seed solution to the fermentation culture medium according to the volume ratio of 10% for culture; after the culture is finished, adding absolute ethyl alcohol, standing for precipitation, collecting precipitations, and drying to obtain the azotobacter chroococcum biological flocculant. According to the azotobacter chroococcum biological flocculant, the azotobacter chroococcum is adopted as a flocculating material, flocculating components mainly include extracellular polymer polysaccharide, viscous capsular polysaccharide and organic acids, amino acids and the like which are generated in a fermentation process, so that the flocculation effect is obvious, and the safety is higher than that of the traditional flocculant; fermentation liquor and thalli are the components of the azotobacter chroococcum without being subjected to solid-liquid separation, so that the separation cost is reduced; and short culture period and good heat stability are achieved.
Description
Technical field
the present invention relates to a kind of flocculation agent, be specifically related to a kind of azotobacter chroococcum biological flocculant and preparation method thereof.
Background technology
Flocculation agent claims again sinking agent, is that a class can make in liquid the not solid suspended particle of free settling (particle diameter 10
-3～10
-7the material of cm) cohesion, precipitation, has been widely used in sanitary sewage disposal, to industry wastewater treatments such as water treatments, papermaking, fermentation, chemical industry, building materials and reduce the fields such as sludge bulking.The flocculation agent being applied at present in water treatment mainly contains two large classes: inorganic salts (as Tai-Ace S 150, iron(ic) chloride etc.) and polymkeric substance thereof (as polymerize aluminum chloride, polyaluminium sulfate or iron etc.); The macromolecular compound (as polyacrylamide) of organic synthesis.Inorganic salt flocculation agent has certain corrodibility, and excess intake aluminium salt pair people has toxicity; The monomer of polyacrylamide flocculation agent is poisonous, easily carcinogenic, and its residue is difficult for being biodegradable, and easily causes secondary pollution, therefore makes its application be very limited.
Microbial flocculant (MBF) is the material with flocculation activity of a class by microorganisms, main chemical compositions is the macromolecular substance such as glycoprotein, polysaccharide, protein, Mierocrystalline cellulose, nucleic acid, because its specific structure and composition has formed good flocculation sediment performance, mainly by thalline adsorb, the outer polymer of born of the same parents flocculate and metabolism in the organic acid that produces and amino acid and the principle such as heavy metal ion toxicity, in plaing in the middle of the particle, the effect such as catch with electric charge, absorption, bridging, net, thereby make elimination of colloid stability, flocculation sediment.Compared with traditional flocculant, microbial flocculant has advantages of safe, nontoxic, biodegradable, non-secondary pollution.But the cultivation of general microorganism needs nitrogenous source, to cultivate cost high, and make fermented liquid color darker after adding nitrogenous source, impact is in the application of water treatment; In addition, the at present microbial flocculant of research or be microorganism fermentating metabolism product, or be thalline itself therefore all needs to adopt centrifugal or filtration to carry out solid-liquid separation after fermentation, has increased the cost of product; In addition the activity of Institute of Micro-biology's produce flocculant is not high, and above reason is restricted applying of microbial flocculant.
Summary of the invention
The object of this invention is to provide azotobacter chroococcum biological flocculant that a kind of culture cycle is shorter, production cost is lower and preparation method thereof.
The technical solution adopted in the present invention is:
The preparation method of azotobacter chroococcum biological flocculant, is characterized in that:
Realized by following steps:
Step 1: configuration fermention medium:
Fermentative medium formula is: N.F,USP MANNITOL 10.0g, KH
2pO
40.2 g, MgSO
47H
2o 0.2g, NaCl 0.2 g, CaCO
35.0g, CaSO
42H
2o 0.1g, adjusts pH to 6.8-7.0, and the 1000mL that adds water mixes, and is sub-packed in 250 mL triangular flasks, every bottled liquid 100 mL, 121 DEG C of sterilizings 30 minutes;
Step 2: the cultivation of fermentation seed liquid:
Get activated azotobacter chroococcum solid slant strains, be seeded to fermention medium, 28-30 DEG C of shaking speed 160rpm fermentation culture 30 hours, obtains fermentation seed liquid;
Step 3: the fermentation culture of flocculation agent:
Get fermentation seed liquid and be seeded in fermention medium by 10% volume ratio, 28-30 DEG C of shaking speed 160rpm fermentation culture 50-60 hour;
Step 4: the preparation of flocculation agent:
After fermentation ends, in fermented liquid, stir and add the dehydrated alcohol of 3 times of volumes, to final concentration be 75%, leave standstill and place 24 hours, wait to precipitate and separate out completely, centrifugal with 4000rpm, collecting precipitation, 50 DEG C of-60 DEG C of oven dry, the faint yellow solid obtaining is azotobacter chroococcum biological flocculant.
The prepared azotobacter chroococcum biological flocculant of preparation method of described azotobacter chroococcum biological flocculant.
The present invention has the following advantages:
The present invention adopts azotobacter chroococcum as flocculation material, its flocculation composition is mainly the outer polymer polysaccharide of born of the same parents, organic acid, amino acid etc. that viscosity capsular polysaccharide and fermenting process produce, these compositions are biodegradable, use safe, nontoxic, non-secondary pollution, flocculating effect and security are all apparently higher than traditional inorganic flocculating agent and organic polymer coargulator.
The azotobacter chroococcum that the present invention uses can be from growing nitrogen-fixing, and culturing process does not need nitrogenous source; Fermented liquid and thalline are flocculation agent composition, do not need solid-liquid separation after fermentation, reduce separation costs, and flocculation agent productive rate is high; Fermentation culture only needs 50-60 hour, and culture cycle is short,, cost-saving.
Flocculation agent main component prepared by the present invention is polysaccharide, Heat stability is good, and 100 DEG C of heating are after 30 minutes, and flocculation agent activity does not reduce.
Brief description of the drawings
Fig. 1 is flocculation agent heat stability test result, and transverse axis is the time, and the longitudinal axis is flocculating rate.
Embodiment
Below in conjunction with embodiment, the present invention will be described in detail.
The preparation method of azotobacter chroococcum biological flocculant involved in the present invention, is realized by following steps:
Step 1: configuration fermention medium:
Fermentative medium formula adopts Ah Xu shellfish fixed nitrogen culture medium prescription, and concrete formula is: N.F,USP MANNITOL 10.0g, KH
2pO
40.2 g, MgSO
47H
2o 0.2g, NaCl 0.2 g, CaCO
35.0g, CaSO
42H
2o 0.1g, adjusts pH to 6.8-7.0, and the 1000mL that adds water mixes, and is sub-packed in 250 mL triangular flasks, every bottled liquid 100 mL, 121 DEG C of sterilizings 30 minutes.
While needing solid medium to cultivate, aforesaid liquid substratum is added to agar by 1.8% volume ratio.
Step 2: the cultivation of fermentation seed liquid:
The solid slant strains of getting activated azotobacter chroococcum (being purchased from Chinese common micro-organisms culture presevation administrative center (CGMCC)), is seeded to fermention medium, and 28-30 DEG C of shaking speed 160rpm fermentation culture 30 hours, obtains fermentation seed liquid.
Step 3: the fermentation culture of flocculation agent:
Get fermentation seed liquid and be seeded in fermention medium by 10% volume ratio, 28-30 DEG C of shaking speed 160rpm fermentation culture 50-60 hour.
Step 4: the preparation of flocculation agent:
After fermentation ends, in fermented liquid, stir and add the dehydrated alcohol of 3 times of volumes, to final concentration be 75%, leave standstill and place 24 hours, wait to precipitate and separate out completely, centrifugal with 4000rpm, collecting precipitation, 50 DEG C of-60 DEG C of oven dry, the faint yellow solid obtaining is azotobacter chroococcum biological flocculant.
Embodiment 1:
Step 1: configuration fermention medium:
Fermentative medium formula adopts Ah Xu shellfish fixed nitrogen culture medium prescription, and concrete formula is: N.F,USP MANNITOL 10.0g, KH
2pO
40.2 g, MgSO
47H
2o 0.2g, NaCl 0.2 g, CaCO
35.0g, CaSO
42H
2o 0.1g, adjusts pH to 6.8, and the 1000mL that adds water mixes, and is sub-packed in 250 mL triangular flasks, every bottled liquid 100 mL, 121 DEG C of sterilizings 30 minutes.
While needing solid medium to cultivate, aforesaid liquid substratum is added to agar by 1.8% volume ratio.
Step 2: the cultivation of fermentation seed liquid:
The solid slant strains of getting activated azotobacter chroococcum, is seeded to fermention medium, and 28 DEG C of shaking speed 160rpm fermentation culture 30 hours, obtain fermentation seed liquid.
Step 3: the fermentation culture of flocculation agent:
Get fermentation seed liquid and be seeded in fermention medium by 10% volume ratio, 28 DEG C of shaking speed 160rpm fermentation culture 50 hours.
Step 4: the preparation of flocculation agent:
After fermentation ends, in 1L fermented liquid, stir and add 3L dehydrated alcohol, making ethanol final concentration is 75%, leaves standstill and places 24 hours, waits to precipitate and separates out completely, centrifugal with 4000 rpm, collecting precipitation, and 50 DEG C of oven dry, obtain 6.5 grams of azotobacter chroococcum biological flocculants.
Embodiment 2:
Step 1: configuration fermention medium:
Fermentative medium formula adopts Ah Xu shellfish fixed nitrogen culture medium prescription, and concrete formula is: N.F,USP MANNITOL 10.0g, KH
2pO
40.2 g, MgSO
47H
2o 0.2g, NaCl 0.2 g, CaCO
35.0g, CaSO
42H
2o 0.1g, adjusts pH to 6.9, and the 1000mL that adds water mixes, and is sub-packed in 250 mL triangular flasks, every bottled liquid 100 mL, 121 DEG C of sterilizings 30 minutes.
While needing solid medium to cultivate, aforesaid liquid substratum is added to agar by 1.8% volume ratio.
Step 2: the cultivation of fermentation seed liquid:
The solid slant strains of getting activated azotobacter chroococcum, is seeded to fermention medium, and 29 DEG C of shaking speed 160rpm fermentation culture 30 hours, obtain fermentation seed liquid.
Step 3: the fermentation culture of flocculation agent:
Get fermentation seed liquid and be seeded in fermention medium by 10% volume ratio, 29 DEG C of shaking speed 160rpm fermentation culture 55 hours.
Step 4: the preparation of flocculation agent:
After fermentation ends, in 1L fermented liquid, stir and add 3L dehydrated alcohol, to final concentration be 75%, leave standstill and place 24 hours, treat that precipitation separates out completely, centrifugal with 4000rpm, collecting precipitation, 55 DEG C of oven dry, obtain 5.6 grams of azotobacter chroococcum biological flocculants.
Embodiment 3:
Step 1: configuration fermention medium:
Fermentative medium formula adopts Ah Xu shellfish fixed nitrogen culture medium prescription, and concrete formula is: N.F,USP MANNITOL 10.0g, KH
2pO
40.2 g, MgSO
47H
2o 0.2g, NaCl 0.2 g, CaCO
35.0g, CaSO
42H
2o 0.1g, adjusts pH to 7.0, and the 1000mL that adds water mixes, and is sub-packed in 250 mL triangular flasks, every bottled liquid 100 mL, 121 DEG C of sterilizings 30 minutes.
While needing solid medium to cultivate, aforesaid liquid substratum is added to agar by 1.8% volume ratio.
Step 2: the cultivation of fermentation seed liquid:
The solid slant strains of getting activated azotobacter chroococcum, is seeded to fermention medium, and 30 DEG C of shaking speed 160rpm fermentation culture 30 hours, obtain fermentation seed liquid.
Step 3: the fermentation culture of flocculation agent:
Get fermentation seed liquid and be seeded in fermention medium by 10% volume ratio, 30 DEG C of shaking speed 160rpm fermentation culture 60 hours.
Step 4: the preparation of flocculation agent:
After fermentation ends, in 1L fermented liquid, stir and add 3L dehydrated alcohol, to final concentration be 75%, leave standstill and place 24 hours, treat that precipitation separates out completely, centrifugal with 4000rpm, collecting precipitation, 60 DEG C of oven dry, obtain 6.2 grams of azotobacter chroococcum biological flocculants.
Flocculation activity is measured the kaolin method that adopts.Get the Kaolin clay suspension of 95 mL 0.3% in 250mL triangular flask, adding distil water is to 95mL, then adds 3 mL1%CaCl
2solution and 2mL 0.5% flocculant solution, shake up, and adjusts pH to 7.0, leaves standstill 3min after jog 1min, gets its supernatant liquor, the OD value with 752 spectrophotometric determination supernatants when the wavelength 550nm, and the while tests as blank using distilled water replacement nutrient solution.
Flocculating rate calculation formula: flocculating rate (%)=(A-B)/A × 100%
In formula: the A-contrast supernatant liquor 550nm OD of place value, the B-sample supernatant liquor 550nm OD of place value.
The flocculation activity of the prepared azotobacter chroococcum biological flocculant of the present invention is higher, its flocculating effect, apparently higher than traditional inorganic flocculating agent and organic polymer coargulator, sees table middle azotobacter chroococcum biological flocculant of the present invention and the comparison of traditional flocculant flocculating effect.
It is cited that content of the present invention is not limited to embodiment, and the conversion of any equivalence that those of ordinary skill in the art take technical solution of the present invention by reading specification sheets of the present invention, is claim of the present invention and contains.
Claims (2)
1. the preparation method of azotobacter chroococcum biological flocculant, is characterized in that:
Realized by following steps:
Step 1: configuration fermention medium:
Fermentative medium formula is: N.F,USP MANNITOL 10.0g, KH
2pO
40.2 g, MgSO
47H
2o 0.2g, NaCl 0.2 g, CaCO
35.0g, CaSO
42H
2o 0.1g, adjusts pH to 6.8-7.0, and the 1000mL that adds water mixes, and is sub-packed in 250 mL triangular flasks, every bottled liquid 100 mL, 121 DEG C of sterilizings 30 minutes;
Step 2: the cultivation of fermentation seed liquid:
Get activated azotobacter chroococcum solid slant strains, be seeded to fermention medium, 28-30 DEG C of shaking speed 160rpm fermentation culture 30 hours, obtains fermentation seed liquid;
Step 3: the fermentation culture of flocculation agent:
Get fermentation seed liquid and be seeded in fermention medium by 10% volume ratio, 28-30 DEG C of shaking speed 160rpm fermentation culture 50-60 hour;
Step 4: the preparation of flocculation agent:
After fermentation ends, in fermented liquid, stir and add the dehydrated alcohol of 3 times of volumes, to final concentration be 75%, leave standstill and place 24 hours, wait to precipitate and separate out completely, centrifugal with 4000rpm, collecting precipitation, 50 DEG C of-60 DEG C of oven dry, the faint yellow solid obtaining is azotobacter chroococcum biological flocculant.
2. the prepared azotobacter chroococcum biological flocculant of the preparation method of azotobacter chroococcum biological flocculant according to claim 1.
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| CN104176803B CN104176803B (en) | 2016-01-13 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104628149A (en) * | 2014-12-23 | 2015-05-20 | 张琰 | Composite flocculant for treating slaughter wastewater |
| CN109423456A (en) * | 2017-08-30 | 2019-03-05 | 中国石油化工股份有限公司 | A kind of azotobacter chroococcum and its identification method and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101298599A (en) * | 2008-06-23 | 2008-11-05 | 中国科学院南海海洋研究所 | Mangrove rhizosphere azotobacter (DZY-N33) and use thereof |
| CN102061313A (en) * | 2010-11-11 | 2011-05-18 | 王紫 | Production method of bioflocculant fermentation liquor and flocculant special for drinking water during flood fighting and disaster relieving and application thereof |
| CN103232951A (en) * | 2013-02-27 | 2013-08-07 | 重庆绿色智能技术研究院 | Enterobacter gergoviae and its use in bioflocculation |
-
2014
- 2014-08-21 CN CN201410412875.2A patent/CN104176803B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101298599A (en) * | 2008-06-23 | 2008-11-05 | 中国科学院南海海洋研究所 | Mangrove rhizosphere azotobacter (DZY-N33) and use thereof |
| CN102061313A (en) * | 2010-11-11 | 2011-05-18 | 王紫 | Production method of bioflocculant fermentation liquor and flocculant special for drinking water during flood fighting and disaster relieving and application thereof |
| CN103232951A (en) * | 2013-02-27 | 2013-08-07 | 重庆绿色智能技术研究院 | Enterobacter gergoviae and its use in bioflocculation |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104628149A (en) * | 2014-12-23 | 2015-05-20 | 张琰 | Composite flocculant for treating slaughter wastewater |
| CN109423456A (en) * | 2017-08-30 | 2019-03-05 | 中国石油化工股份有限公司 | A kind of azotobacter chroococcum and its identification method and application |
| CN109423456B (en) * | 2017-08-30 | 2022-08-19 | 中国石油化工股份有限公司 | Azotobacter chroococcum as well as identification method and application thereof |
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| Publication number | Publication date |
|---|---|
| CN104176803B (en) | 2016-01-13 |
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