CN110129375A - A kind of preparation method of microbial flocculant - Google Patents
A kind of preparation method of microbial flocculant Download PDFInfo
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- CN110129375A CN110129375A CN201910402054.3A CN201910402054A CN110129375A CN 110129375 A CN110129375 A CN 110129375A CN 201910402054 A CN201910402054 A CN 201910402054A CN 110129375 A CN110129375 A CN 110129375A
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 230000003311 flocculating effect Effects 0.000 claims abstract description 64
- 238000000034 method Methods 0.000 claims abstract description 63
- 230000000694 effects Effects 0.000 claims abstract description 57
- 238000005189 flocculation Methods 0.000 claims abstract description 50
- 230000016615 flocculation Effects 0.000 claims abstract description 50
- 238000011534 incubation Methods 0.000 claims abstract description 31
- 239000011265 semifinished product Substances 0.000 claims abstract description 16
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims abstract description 15
- 238000000605 extraction Methods 0.000 claims abstract description 14
- 239000001963 growth medium Substances 0.000 claims abstract description 5
- 239000006228 supernatant Substances 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 25
- 239000002609 medium Substances 0.000 claims description 25
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 239000000243 solution Substances 0.000 claims description 19
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 16
- 230000001580 bacterial effect Effects 0.000 claims description 16
- 238000010790 dilution Methods 0.000 claims description 16
- 239000012895 dilution Substances 0.000 claims description 16
- 239000005995 Aluminium silicate Substances 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 235000012211 aluminium silicate Nutrition 0.000 claims description 14
- 239000002689 soil Substances 0.000 claims description 14
- 238000002835 absorbance Methods 0.000 claims description 12
- 239000010802 sludge Substances 0.000 claims description 12
- 238000012216 screening Methods 0.000 claims description 11
- 239000000725 suspension Substances 0.000 claims description 11
- 229910052622 kaolinite Inorganic materials 0.000 claims description 10
- 230000001376 precipitating effect Effects 0.000 claims description 10
- 238000000746 purification Methods 0.000 claims description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 8
- 238000005303 weighing Methods 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 5
- 239000013049 sediment Substances 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 2
- 239000002244 precipitate Substances 0.000 claims description 2
- 238000011218 seed culture Methods 0.000 claims description 2
- 238000001291 vacuum drying Methods 0.000 claims description 2
- 238000002242 deionisation method Methods 0.000 claims 1
- 150000002576 ketones Chemical class 0.000 claims 1
- 244000005700 microbiome Species 0.000 abstract description 14
- 239000000047 product Substances 0.000 abstract description 7
- 238000002481 ethanol extraction Methods 0.000 abstract description 6
- 125000005211 alkyl trimethyl ammonium group Chemical group 0.000 abstract description 4
- 238000003811 acetone extraction Methods 0.000 abstract description 3
- 238000009629 microbiological culture Methods 0.000 abstract description 3
- 238000000855 fermentation Methods 0.000 description 9
- 230000004151 fermentation Effects 0.000 description 9
- 241000894006 Bacteria Species 0.000 description 8
- 238000005119 centrifugation Methods 0.000 description 8
- 230000007423 decrease Effects 0.000 description 7
- 230000004060 metabolic process Effects 0.000 description 6
- 238000007747 plating Methods 0.000 description 6
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- 239000008394 flocculating agent Substances 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 4
- 229910052782 aluminium Inorganic materials 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000002351 wastewater Substances 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000012043 crude product Substances 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 229960004756 ethanol Drugs 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 241000186046 Actinomyces Species 0.000 description 2
- 208000014644 Brain disease Diseases 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 208000032274 Encephalopathy Diseases 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241000235342 Saccharomycetes Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000004411 aluminium Substances 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- -1 iron ion Chemical class 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000004065 wastewater treatment Methods 0.000 description 2
- 102100030310 5,6-dihydroxyindole-2-carboxylic acid oxidase Human genes 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 101000773083 Homo sapiens 5,6-dihydroxyindole-2-carboxylic acid oxidase Proteins 0.000 description 1
- 206010029350 Neurotoxicity Diseases 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 241000187561 Rhodococcus erythropolis Species 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 description 1
- 159000000013 aluminium salts Chemical class 0.000 description 1
- 229910000329 aluminium sulfate Inorganic materials 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- SWLVFNYSXGMGBS-UHFFFAOYSA-N ammonium bromide Chemical compound [NH4+].[Br-] SWLVFNYSXGMGBS-UHFFFAOYSA-N 0.000 description 1
- 239000011449 brick Substances 0.000 description 1
- 230000031709 bromination Effects 0.000 description 1
- 238000005893 bromination reaction Methods 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000701 coagulant Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000000909 electrodialysis Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000003317 industrial substance Substances 0.000 description 1
- 239000010842 industrial wastewater Substances 0.000 description 1
- 239000008235 industrial water Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000007269 microbial metabolism Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000007135 neurotoxicity Effects 0.000 description 1
- 231100000228 neurotoxicity Toxicity 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- 231100000419 toxicity Toxicity 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Mycology (AREA)
- Environmental & Geological Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Hydrology & Water Resources (AREA)
- Water Supply & Treatment (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Separation Of Suspended Particles By Flocculating Agents (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to the technical fields of microorganism, and disclose a kind of preparation method of microbial flocculant, by setting different incubation times, it is 0~22h in incubation time, culture flocculation activity increases with incubation time to be improved, the flocculating rate highest when cultivating 66h, the flocculating rate of culture solution starts to reduce after 66h, to make user's to obtain the culture flocculation activity most suitable time, the product quality of user is effectively promoted, by to acetone extraction, the comparison of 3 kinds of extracting methods of ethanol extraction method and cetyl trimethylammonium bromide extraction method shows, cetyl trimethylammonium bromide method yield with higher, the dried product of 1.33g can be obtained in every liter of culture solution, cetyl trimethylammonium bromide method is relatively high, it is 71.8%, effectively improve the activity of flocculant semifinished product, pass through ten The flocculant yield that six alkyl trimethyl ammonium bromide methods are extracted from microbial culture medium is high, and flocculation activity with higher has boundless application prospect.
Description
Technical field
The present invention relates to the technical field of microorganism, specially a kind of preparation method of microbial flocculant.
Background technique
With the growth of rapid development of economy and population, the annual water consumption in China and quantity of wastewater effluent are also increasing,
Sewage handling problem is increasingly serious, and Water purification problem also can not be ignored.Currently, the processing method of water and waste water mainly has life
Change, ion exchange, absorption, chemical oxidation, electrodialysis and flocculation sedimentation etc. are a variety of.Wherein apply most universal, lower-cost place
Reason method is flocculent precipitation.In flocculent precipitation processing waste water, the exploitation selection of flocculant is one of its key technology.Wadding
Solidifying agent is not only applicable to Industrial Wastewater Treatment, while being also widely used in fermenting as a kind of widely used water process reagent
Treatment of Industrial Water and chemical industry, food, medicine and other fields separation of solid and liquid process.Water supply and wastewater treatment are normal for a long time
Flocculant is mainly using aluminium salt, molysite and its polymer as the inorganic flocculating agent of representative and having by representative of polyacrylamide
Machine high polymer coagulant.Although these flocculants have preferable flocculation activity, but their residue is mostly harmful, and there are two
Secondary pollution problem.The health of the mankind is also endangered simultaneously.Since application aluminum salt flocculant method treated sludge is answered as fertilizer
After agricultural, or landfill, the aluminium content in soil is caused to increase, so that plant aluminium evil is occurred, normally given birth to influence plant
It is long or even dead；Simultaneous crops enter food chain, and the final health for influencing the mankind causes aluminum encephalopathy.It is senile
Dementia is exactly one kind of aluminum encephalopathy.Iron flocculant not only has strong corrosivity, the use of limiting device, Er Qierong
Easily residual iron ion, and it is processed after water have coloration.Most common Syn-Organic flocculants are exactly polypropylene
Amide.Although its no any toxicity itself, its refractory organics easily cause secondary pollution, and the residual of polymer monomer
Not only there is strong neurotoxicity, but also be strong carcinogen.Therefore the use of this kind of flocculant receives restriction.It opens simultaneously
It sends out nontoxic, is harmless, is without secondary pollution, can more and more be paid close attention to biodegradable efficient flocculant by researcher.Micro- life
Object flocculant (MBF, microbialflocculant) is a kind of metabolite generated by microorganism or its secretion, it is
Using microbial technique, by being with Biodegradable and peace obtained by the microbial fermentations such as bacterium, fungi, extraction, purification
The water treatment agent efficient, nontoxic, without secondary pollution of full property.Various polysaccharides, the egg that it is mainly generated by microbial metabolism
White matter or protein and carbohydrate participate in the high-molecular compound to be formed.The microbe species of microbial flocculant can be generated very
More, they are largely present in soil, activated sludge and deposit.
Research to microbial flocculant, what is appeared in the newspapers earliest is that nineteen thirty-five Butterfield is screened from activated sludge
To one plant of bacterium for producing flocculant.Subsequent researcher has continued research in this respect.The seventies J.Nakamura et al. from
Filter out 19 plants of bacterium for producing flocculant strains in 214 plants of bacterium such as mould, bacterium, actinomyces, saccharomycete, including 8 plants of moulds, 5 plants thin
Bacterium, 5 plants of actinomyces and 1 saccharomycete.The flocculating agent A J7002 effect that wherein Aspergzllussoda is generated is best；TakagiH
Deng being separated to from soil one plant of fungi to multiple-microorganism cell and suspension colloid particle with good flocculation
Paecilomycessp.；Late nineteen eighties Kurane et al. one plant of R.erythropolis of separation screening from soil, is made
It is named as the microbial flocculant of NOC-1.NOC 1 is used for livestock products waste water, bulking sludge and the production of brick field and given up by Kurane et al.
The processing of water etc. achieves good effect, it is considered to be presently found best microbial flocculant.Currently, South Korea
KwonG.S., SeoHyunHyo, KimYoung-Jun et al., German P.Reinhard specification page 1/4
3CN105731657B3 and the N.Levy of Israel et al. have also carried out more comprehensive research to microbial flocculant.[3]
So far researcher has been obtained for the more plants of bacterium for producing flocculant of microbe with different flocculation abilities, these microbial flocculants
Have the advantages that in use it is efficient, safe, free from environmental pollution, the fields such as food, chemistry and pharmacy have it is huge
Potential using value, the problem of existing microbial flocculant maximum is that the activity of microbial flocculant is universal low, thus
Need a kind of microbial flocculant preparation method that flocculant activity can be improved.
Summary of the invention
(1) the technical issues of solving
In view of the deficiencies of the prior art, the present invention provides a kind of preparation method of microbial flocculant, has flocculant
The advantages that activity is high solves the problems, such as that flocculant activity is universal low.
(2) technical solution
To achieve the above object, the invention provides the following technical scheme: a kind of preparation method of microbial flocculant, including
Following steps:
(1) strain samples: after activated sludge dilution, placing it in 30C constant-temperature table, is then coated with using dilution plate
Method obtains single colonie, then will access shaking flask culture in screening and culturing medium by bacterial strain after purification respectively, and culture solution carries out flocculation work later
Property Preliminary Determination, flocculation activity the higher person be produce flocculant bacterial strain.
(2) flocculation activity measures: weighing kaolin, distilled water is therefrom added to kaolinite soil suspension is made；It then takes to be measured
Liquid is added in kaolinite soil suspension, is mixed slowly, and is stood, and supernatant absorbance is measured at 550nm, is replaced with deionized water
Prepare liquid surveys absorbance as control, with method.
(4) Spawn incubation: will carry out again shaking flask culture after strain seed culture, respectively when pH initial to culture medium and culture
Between influence to flocculating rate be measured, the strain after screening is subjected to shaking flask culture, incubation time is respectively 22h, 44h,
66h, 88h survey its flocculating rate result.
(5) extraction of flocculant: medium centrifugal is separated, to remove precipitating, 1.5 times of volumes of people are added in supernatant
Acetone, removes supernatant at this time, and ether washing precipitating is then dried in vacuo, obtains flocculant semifinished product.
(6) flocculant adds: measurement flocculant dosage volume fraction is 0.75%, 1%, 1.25%
Flocculating rate when with 1.5%.
Preferably, the culture solution is centrifuged 10min under 12000r/min revolving speed.
Preferably, the sediment of the broth out is thallus and sundries.
Preferably, supernatant dosage flocculating effect at 1.00%~1.25% is best, reaches 59.5%. and is less than
Or when being greater than this range, flocculating effect decreases.
Preferably, the method that the flocculant extracts is three kinds, respectively acetone method, Ethanol Method and cetyl trimethyl
Ammonium bromide method.
(3) beneficial effect
Compared with prior art, the present invention provides a kind of preparation methods of microbial flocculant, have following beneficial to effect
Fruit:
1, the preparation method of the microbial flocculant, by acetone extraction, ethanol extraction method and cetyl front three
The comparison of base ammonium bromide 3 kinds of extracting methods of method shows that cetyl trimethylammonium bromide method yield with higher, every liter is cultivated
The dried product of 1.33g can be obtained in liquid, and the semifinished product flocculation activity that 3 kinds of methods obtain is not much different, cetyl trimethyl bromination
Ammonium method is relatively high, is 71.8%, effectively improves the activity of flocculant semifinished product, pass through cetyl trimethylammonium bromide
The flocculant yield that method is extracted from microbial culture medium is high, flocculation activity with higher, and not will cause to environment
Secondary pollution has boundless application prospect.
2, the preparation method of the microbial flocculant, by setting different incubation time, measure its flocculating rate as a result,
It is 0~22h in incubation time, rapidly, metabolism is vigorous for microorganism growth, and culture flocculation activity increases with incubation time to be improved,
22~66h flocculating rate slightly increases, but changes less, the flocculating rate highest when cultivating 66h, since microorganism grows generation after 66h
The consumption thanked, the flocculating rate of culture solution start to reduce, to make user's to obtain the culture flocculation activity most suitable time, effectively
Ground improves the product quality of user.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical solution in the embodiment of the present invention is clearly and completely retouched
It states, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the present invention
In embodiment, every other implementation obtained by those of ordinary skill in the art without making creative efforts
Example, shall fall within the protection scope of the present invention.
Embodiment one:
A kind of preparation method of microbial flocculant, comprising the following steps:
(1) strain samples: after activated sludge dilution, placing it in 30C constant-temperature table, 160r/min enrichment culture three
It, then obtains single colonie using dilution plate rubbing method, then bacterial strain will connect in people's 100mL screening and culturing medium after purification respectively,
With 30 DEG C, 160r/min shaking flask culture 3 days, culture solution carries out the Preliminary Determination of flocculation activity later, flocculation activity the higher person
To produce flocculant bacterial strain.
(2) flocculant activity measures: weighing 0.1g kaolin, distilled water is therefrom added to 200mL, it is outstanding that kaolin is made
Liquid takes 2mL prepare liquid to be added in 200mL kaolinite soil suspension, mixes slowly 10min, then stand 5min, surveys at 550nm
Measure supernatant absorbance B, using deionized water replace prepare liquid as control, with method survey absorbance A, flocculating rate=(A-B)/
A.100%.
(3) Spawn incubation: strain is inoculated in after being activated one day in plating medium, strain is inoculated into dress with oese
There is 50mI, the fermented and cultured in the conical flask of fermentation medium, cultivation temperature is 30 DEG C, shaking speed 150r/min, culture
Time is 22h, and supernatant is taken to measure its flocculating rate after being centrifuged, and when incubation time is 0~22h, microorganism growth is rapid,
Be metabolized it is vigorous, culture flocculation activity with incubation time increase improve.
(4) extraction of flocculant: wherein the method for acetone method is medium centrifugal separation (5000r/min, centrifugation
25min), removal precipitates thallus and sundries, and the acetone of 1.5 times of volumes is added in supernatant, and 4 DEG C stand overnight, 12000r/min,
It is centrifuged 10min, supernatant, subsequent ether washing precipitating is removed at this time, then will be dried in vacuo, obtains flocculant semifinished product.
(5) flocculant adds: adding flocculation when flocculant dosage volume is 0.75%, 1%, 1.25% and 1.5%
Rate, supernatant dosage flocculating effect at 1.00%~1.25% is best, reaches 59.5%, when being less than or greater than this range,
Flocculating effect decreases.
Embodiment two:
A kind of preparation method of microbial flocculant, comprising the following steps:
(1) strain samples: after activated sludge dilution, sets and place it in 30C constant-temperature table, 160r/min enrichment culture four
It, then obtains single colonie using dilution plate rubbing method, then bacterial strain will connect in people's 100mL screening and culturing medium after purification respectively,
With 30 DEG C, 160r/min shaking flask culture 4 days, culture solution carries out the Preliminary Determination of flocculation activity later, flocculation activity the higher person
To produce flocculant bacterial strain.
(2) flocculant activity measures: weighing 0.2g kaolin, distilled water is therefrom added to 400mL, it is outstanding that kaolin is made
Liquid takes 2mL prepare liquid to be added in 200mL kaolinite soil suspension, mixes slowly 10min, stands 5min, measures at 550nm
Clear liquid absorbance B replaces prepare liquid as control, with method survey absorbance A, flocculating rate=(A-B)/A.100% using deionized water.
(3) Spawn incubation: strain being inoculated in plating medium and is activated two days later, and strain is inoculated into dress with oese
There is a 50mI, fermented and cultured in the conical flask of fermentation medium, cultivation temperature is 30 DEG C, shaking speed 150r/min, when culture
Between be 44h, take supernatant to measure its flocculating rate after centrifugation, incubation time be 22~66h, flocculating rate slightly increase, but change
Less.
(4) extraction of flocculant: Ethanol Method, medium centrifugal separation (5000r/min is centrifuged 25min), removal precipitating bacterium
Body and sundries in supernatant plus the dehydrated alcohol of 1.5 times of volumes of people pre-cooling, mix centrifugation, collect precipitating, 70% ethanol washing is heavy
It forms sediment, vacuum drying obtains flocculant semifinished product.
(5) flocculant adds: adding flocculation when flocculant dosage volume is 0.75%, 1%, 1.25% and 1.5%
Rate, supernatant dosage flocculating effect at 1.00%~1.25% is best, reaches 59.5%, when being less than or greater than this range,
Flocculating effect decreases.
Embodiment three:
A kind of preparation method of microbial flocculant, comprising the following steps:
(1) strain samples: after activated sludge dilution, sets and place it in 30C constant-temperature table, 160r/min enrichment culture five
It, then obtains single colonie using dilution plate rubbing method, then bacterial strain will connect in people's 100mL screening and culturing medium after purification respectively,
With 30 DEG C, 160r/min shaking flask culture 5 days, culture solution carries out the Preliminary Determination of flocculation activity later, flocculation activity the higher person
To produce flocculant bacterial strain.
(2) flocculant activity measures: weighing 0.3g kaolin, distilled water is therefrom added to 700mL, it is outstanding that kaolin is made
Liquid takes 2mL prepare liquid to be added in 200mL kaolinite soil suspension, mixes slowly 10min, stands 5min, measures at 550nm
Clear liquid absorbance B replaces prepare liquid as control, with method survey absorbance A, flocculating rate=(A-B)/A.100% using deionized water.
(3) Spawn incubation: strain being inoculated in plating medium and is activated two days later, and strain is inoculated into dress with oese
There is a 50mI, fermented and cultured in the conical flask of fermentation medium, cultivation temperature is 30 DEG C, shaking speed 150r/min, when culture
Between be 66h, take supernatant to measure its flocculating rate after centrifugation, incubation time be 0~22h, microorganism growth rapidly, be metabolized it is prosperous
It contains, culture flocculation activity increases with incubation time to be improved, and 22~66h flocculating rate slightly increases, but changes less, is being cultivated
Flocculating rate highest when 66h.
(4) extraction of flocculant: cetyl trimethylammonium bromide method, medium centrifugal separation, removal precipitating thallus and
Sundries, in supernatant slowly plus people's concentration be 10% cetyl trimethylammonium bromide, received after no Precipitation, centrifugation
Collection precipitating, precipitating is dissolved in the NaCl solution of 2mol/L, the dehydrated alcohol of 2 times of volumes is added to be precipitated, and is analysed until without precipitating
Out, it is washed respectively with 75% ethyl alcohol, dehydrated alcohol, ether, is dried in vacuo, obtains flocculant semifinished product.
(5) flocculant adds: adding flocculation when flocculant dosage volume is 0.75%, 1%, 1.25% and 1.5%
Rate, supernatant dosage flocculating effect at 1.00%~1.25% is best, reaches 59.5%, when being less than or greater than this range,
Flocculating effect decreases.
Example IV:
A kind of preparation method of microbial flocculant, comprising the following steps:
(1) strain samples: after activated sludge dilution, placing it in 30C constant-temperature table, 160r/min enrichment culture six
It, then obtains single colonie using dilution plate rubbing method, then bacterial strain will connect in people's 100mL screening and culturing medium after purification respectively,
With 30 DEG C, 160r/min shaking flask culture six days, culture solution carries out the Preliminary Determination of flocculation activity later, flocculation activity the higher person
To produce flocculant bacterial strain.
(2) flocculant activity measures: weighing 0.3g kaolin, distilled water is therefrom added to 800mL, it is outstanding that kaolin is made
Liquid；It takes 2mL prepare liquid to be added in 200mL kaolinite soil suspension, mixes slowly 10min, stand 5min, measured at 550nm
Clear liquid absorbance B replaces prepare liquid as control, with method survey absorbance A, flocculating rate=(A-B)/A.100% using deionized water.
(3) Spawn incubation: strain is inoculated in after being activated four days in plating medium, strain is inoculated into dress with oese
There is a 50mI, fermented and cultured in the conical flask of fermentation medium, cultivation temperature is 30 DEG C, shaking speed 150r/min, when culture
Between be 88h, take supernatant to measure its flocculating rate after centrifugation, incubation time be 0~22h, microorganism growth rapidly, be metabolized it is prosperous
It contains, culture flocculation activity increases with incubation time to be improved, and 22~66h flocculating rate slightly increases, but changes less, is being cultivated
Flocculating rate highest when 66h, due to the consumption of microorganism growth metabolism after 66h, the flocculating rate of culture solution starts to reduce.
(4) extraction of flocculant: in the case where the dosage of flocculant semifinished product is 3mg/L, acetone is mentioned respectively
It follows the example of, the flocculant crude product that ethanol extraction method and cetyl trimethylammonium bromide extraction method obtain carries out the measurement of flocculating rate,
Flocculating rate result is followed successively by 66.0%, 67.4% and 71.8%, and the semifinished product flocculation activity that 3 kinds of methods obtain is not much different, and ten
The flocculation activity that six alkyl trimethyl ammonium bromide methods obtain flocculant is relatively high.
(5) flocculant adds: adding flocculation when flocculant dosage volume is 0.75%, 1%, 1.25% and 1.5%
Rate, supernatant dosage flocculating effect at 1.00%~1.25% is best, reaches 59.5%, when being less than or greater than this range,
Flocculating effect decreases.
Embodiment five:
A kind of preparation method of microbial flocculant, comprising the following steps:
(1) strain samples: after activated sludge dilution, placing it in 30C constant-temperature table, 160r/min enrichment culture six
It, then obtains single colonie using dilution plate rubbing method, then bacterial strain will connect in people's 100mL screening and culturing medium after purification respectively,
With 30 DEG C, 160r/min shaking flask culture six days, culture solution carries out the Preliminary Determination of flocculation activity later, flocculation activity the higher person
To produce flocculant bacterial strain.
(2) flocculant activity measures: weighing 0.4g kaolin, distilled water is therefrom added to 1000mL, it is outstanding that kaolin is made
Liquid takes kaolinite soil suspension, calcium chloride water each 10ml, 0.5ml respectively, is uniformly mixed, therefrom takes 9.8ml, then 0.2ml is taken to send out
Zymotic fluid is added in small beaker, and fermentation liquid is not added in control, and pH value is adjusted in small beaker to 7.5-8.0, in the process with PH meter
It mixes well, pours into test tube, stand half an hour, first observe its Flocculating Effect of Flocculant with observation method, then drawn with dropper
Supernatant surveys its light absorption value into cuvette in ultraviolet specrophotometer at 550nm, and calculates flocculating rate: flocculating rate=(A-
B)/A.100%.
(3) Spawn incubation: strain is inoculated in after being activated four days in plating medium, strain is inoculated into dress with oese
There is a 50mI, fermented and cultured in the conical flask of fermentation medium, cultivation temperature is 30 DEG C, shaking speed 150r/min, when culture
Between be 88h, take supernatant to measure its flocculating rate after centrifugation, incubation time be 0~22h, microorganism growth rapidly, be metabolized it is prosperous
It contains, culture flocculation activity increases with incubation time to be improved, and 22~66h flocculating rate slightly increases, but changes less, is being cultivated
Flocculating rate highest when 66h, due to the consumption of microorganism growth metabolism after 66h, the flocculating rate of culture solution starts to reduce.
(4) extraction of flocculant: in the case where the dosage of flocculant semifinished product is 3mg/L, acetone is mentioned respectively
It follows the example of, the flocculant crude product that ethanol extraction method and cetyl trimethylammonium bromide extraction method obtain carries out the measurement of flocculating rate,
Flocculating rate result is followed successively by 66.0%, 67.4% and 71.8%, and the semifinished product flocculation activity that 3 kinds of methods obtain is not much different, and ten
The flocculation activity that six alkyl trimethyl ammonium bromide methods obtain flocculant is relatively high.
(5) flocculant adds: adding flocculation when flocculant dosage volume is 0.75%, 1%, 1.25% and 1.5%
Rate, supernatant dosage flocculating effect at 1.00%~1.25% is best, when reaching 59.5%. less than or greater than this range,
Flocculating effect decreases.
Embodiment six:
A kind of preparation method of microbial flocculant, comprising the following steps:
(1) strain samples: after activated sludge dilution, placing it in 30C constant-temperature table, 160r/min enrichment culture six
It, then obtains single colonie using dilution plate rubbing method, then bacterial strain will connect in people's 100mL screening and culturing medium after purification respectively,
With 30 DEG C, 160r/min shaking flask culture seven days, culture solution carries out the Preliminary Determination of flocculation activity later, flocculation activity the higher person
To produce flocculant bacterial strain.
(2) flocculant activity measures: weighing 0.4g kaolin, distilled water is therefrom added to 1000mL, it is outstanding that kaolin is made
Liquid takes kaolinite soil suspension, calcium chloride water each 10ml, 0.5ml respectively, is uniformly mixed, therefrom takes 9.8ml, then 0.2ml is taken to send out
Zymotic fluid is added in small beaker, and fermentation liquid is not added in control, and pH value is adjusted in small beaker to 7.5-8.0, in the process with PH meter
It mixes well, pours into test tube, stand half an hour, first observe its Flocculating Effect of Flocculant with observation method, then drawn with dropper
Supernatant surveys its light absorption value into cuvette in ultraviolet specrophotometer at 550nm, and calculates flocculating rate: flocculating rate=(A-
B)/A.100%.
(3) Spawn incubation: strain is inoculated in after being activated five days in plating medium, strain is inoculated into dress with oese
There is a 50mI, fermented and cultured in the conical flask of fermentation medium, cultivation temperature is 30 DEG C, shaking speed 150r/min, when culture
Between be 88h, take supernatant to measure its flocculating rate after centrifugation, incubation time be 0~22h, microorganism growth rapidly, be metabolized it is prosperous
It contains, culture flocculation activity increases with incubation time to be improved, and 22~66h flocculating rate slightly increases, but changes less, is being cultivated
Flocculating rate highest when 66h, due to the consumption of microorganism growth metabolism after 66h, the flocculating rate of culture solution starts to reduce.
(4) extraction of flocculant: in the case where the dosage of flocculant semifinished product is 4mg/L, acetone is mentioned respectively
It follows the example of, the flocculant crude product that ethanol extraction method and cetyl trimethylammonium bromide extraction method obtain carries out the measurement of flocculating rate,
Flocculating rate result is followed successively by 66.0%, 67.4% and 71.8%, and the semifinished product flocculation activity that 3 kinds of methods obtain is not much different, and ten
The flocculation activity that six alkyl trimethyl ammonium bromide methods obtain flocculant is relatively high.
(5) flocculant adds: adding flocculation when flocculant dosage volume is 0.75%, 1%, 1.25% and 1.5%
Rate, supernatant dosage flocculating effect at 1.00%~1.25% is best, when reaching 59.5%. less than or greater than this range,
Flocculating effect decreases.
Judgment criteria: being qualification when flocculant activity is detected as percent 60 or more.
The beneficial effects of the present invention are: by acetone extraction, ethanol extraction method and cetyl trimethylammonium bromide
The comparison of 3 kinds of extracting methods of extraction method shows that cetyl trimethylammonium bromide method yield with higher, every liter of culture solution can
The dried product of 1.33g is obtained, the semifinished product flocculation activity that 3 kinds of methods obtain is not much different, cetyl trimethylammonium bromide method
It is relatively high, it is 71.8%, effectively improves the activity of flocculant semifinished product, mentioned from microbial culture medium by CATB method
The flocculant yield obtained is high, flocculation activity with higher, and not will cause secondary pollution to environment, has boundless
Application prospect, by setting different incubation time, measure its flocculating rate as a result, being 0~22h, micro- life in incubation time
Rapidly, metabolism is vigorous for object growth, and culture flocculation activity increases with incubation time to be improved, and 22~66h flocculating rate slightly increases, but
Variation is little, the flocculating rate highest when cultivating 66h, and due to the consumption of microorganism growth metabolism after 66h, the flocculating rate of culture solution is opened
Begin to reduce, to make user's to obtain the culture flocculation activity most suitable time, the product quality of user is effectively promoted.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (5)
1. a kind of preparation method of microbial flocculant, which comprises the following steps:
(1) strain samples: after activated sludge dilution, placing it in 30C constant-temperature table, is then obtained using dilution plate rubbing method
Single colonie is obtained, then shaking flask culture in screening and culturing medium will be accessed by bacterial strain after purification respectively, culture solution carries out flocculation activity later
Preliminary Determination, flocculation activity the higher person are to produce flocculant bacterial strain.
(2) flocculation activity measures: weighing the kaolin of 0.1~0.5g, distilled water is therefrom added to kaolinite soil suspension is made, then
It takes prepare liquid to be added in kaolinite soil suspension, mixes slowly, stand, and measure supernatant absorbance at 550nm, with deionization
Water replaces prepare liquid as control, surveys absorbance with method.
(3) Spawn incubation: shaking flask culture will be carried out again after strain seed culture, respectively pH initial to culture medium and incubation time pair
The influence of flocculating rate is measured, and the strain after screening is carried out shaking flask culture, and incubation time is respectively 22h, 44h, 66h,
88h surveys its flocculating rate result.
(4) extraction of flocculant: medium centrifugal is separated, to remove precipitating, the third of 1.5 times of volumes of people is added in supernatant
Ketone removes supernatant at this time, then washs ether and precipitates, and vacuum drying obtains flocculant semifinished product.
(5) flocculant adds: wadding when measurement flocculant dosage volume fraction is 0.75%, 1%, 1.25% and 1.5%
Solidifying rate.
2. a kind of preparation method of microbial flocculant according to claim 1, it is characterised in that: the culture solution in
10min is centrifuged under 12000r/min revolving speed.
3. a kind of preparation method of microbial flocculant according to claim 1, it is characterised in that: the broth out
Sediment be thallus and sundries.
4. a kind of preparation method of microbial flocculant according to claim 1, it is characterised in that: the supernatant adds
Amount flocculating effect at 1.00%~1.25% is best, reaches 59.5%.
5. a kind of preparation method of microbial flocculant according to claim 1, it is characterised in that: the flocculant extracts
Method be three kinds, respectively acetone method, Ethanol Method and cetyl trimethylammonium bromide method.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050282202A1 (en) * | 2004-05-21 | 2005-12-22 | Brolaski Mark N | Kits and processes for removing contaminants from nucleic acids in environmental and biological samples |
| CN102399823A (en) * | 2010-09-09 | 2012-04-04 | 上海海帝园艺有限公司 | Method for extracting bioflocculant |
| CN104726518A (en) * | 2015-04-09 | 2015-06-24 | 哈尔滨工业大学 | Production method for microbial flocculants |
| CN110054361A (en) * | 2019-05-13 | 2019-07-26 | 辽宁石油化工大学 | A kind of electric field-enhanced MBR sewage treatment process |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050282202A1 (en) * | 2004-05-21 | 2005-12-22 | Brolaski Mark N | Kits and processes for removing contaminants from nucleic acids in environmental and biological samples |
| CN102399823A (en) * | 2010-09-09 | 2012-04-04 | 上海海帝园艺有限公司 | Method for extracting bioflocculant |
| CN104726518A (en) * | 2015-04-09 | 2015-06-24 | 哈尔滨工业大学 | Production method for microbial flocculants |
| CN110054361A (en) * | 2019-05-13 | 2019-07-26 | 辽宁石油化工大学 | A kind of electric field-enhanced MBR sewage treatment process |
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Application publication date: 20190816 |