CN106630063A - Microbial flocculating agent generated by paenibacillus and applications thereof - Google Patents
Microbial flocculating agent generated by paenibacillus and applications thereof Download PDFInfo
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- CN106630063A CN106630063A CN201611186231.1A CN201611186231A CN106630063A CN 106630063 A CN106630063 A CN 106630063A CN 201611186231 A CN201611186231 A CN 201611186231A CN 106630063 A CN106630063 A CN 106630063A
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
- C02F1/5236—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using inorganic agents
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
- C02F1/5236—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using inorganic agents
- C02F1/5245—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using inorganic agents using basic salts, e.g. of aluminium and iron
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
- C02F1/54—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using organic material
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- 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
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Separation Of Suspended Particles By Flocculating Agents (AREA)
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- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention discloses a microbial flocculating agent generated by paenibacillus and applications thereof. The microbial flocculating agent is generated by paenibacillus sp. LX03, and is prepared by the following steps: inoculating paenibacillus sp. LX03 into a fermentation culture medium, carrying out cultivation on a shaking table for 2 to 3 days, subjecting the fermentation broth to centrifugation to remove bacterial cells, adding ethanol, which is 2 to 3 times the volume of the fermentation broth, to precipitate extracellular products, drying the extracellular products, and adding water to dissolve extracellular products to obtain the microbial flocculating agent. When the microbial flocculating agent is compounded with iron ions and aluminum ions, a good flocculating effect can be achieved. When the microbial flocculating agent is used with an inorganic flocculating agent namely polymeric aluminum chloride, the using amount of polymeric aluminum chloride is largely reduced, and a better flocculating effect is formed in a short time. The microbial flocculating agent can be used as a good water treatment flocculating agent and flocculating aid, and can be widely used in the fields such as household wastewater treatment, industrial wastewater treatment, and the like.
Description
Technical field
The invention belongs to microbial flocculant water-treatment technology field, and in particular to micro- life that a Bacillus species are produced
Thing flocculant and its application.
Background technology
A large amount of discharges of sanitary sewage and trade effluent produce severe contamination to environment, affect the ecological balance.Flocculant with
The pollutants such as pigment, solid suspended particle, heavy metal ion in sewage can be combined, and form larger floccule body further
Sedimentation, convenient that various pollutants are focused on, Jing after flocculant process, the water quality of sewage is purified.At present, flocculate
Agent substantially can be divided three classes：Inorganic flocculating agent, including molysite, aluminium salt, aluminium polychloride, poly-ferric chloride etc.；It is artificial synthesized
Organic polymer coargulator, such as polyacrylamide and its derivative；Natural biological flocculant, such as microbial flocculant.
Natural microbial flocculant is that the metabolism that the class that microorganism produces during the fermentation has flocculating function is produced
Thing.Microbial flocculant is made up of polysaccharide, protein, glycoprotein and nucleic acid etc., wherein again in the majority with polysaccharide and play main work
Not degradable solid suspended particle and colloidal particle aggegation sedimentation can be made with, microbial flocculant, is imitated with good flocculation
Really.Microbial flocculant has the advantages that nontoxic, safe and efficient, biodegradable, but such as document 1 (Gan Li, Meng Zhaoping, micro- life
The progress of thing flocculant, water technology, 2006,32 (4):It is 5-9) described, consumption is big, high cost the problems such as seriously restrict
The production of microbial flocculant and application.At present, microbial flocculant and other flocculants are carried out into compounding use, is microorganism
One direction of flocculant research, many studies have shown that, the compounding use of microbial flocculant can not only improve flocculating effect, also
Can be greatly reduced the dosage of flocculant, such as document 2 (Huang Jing, Yang Chaohui, grandson's Pei stones, microbial flocculant be polymerized
The response surface optimization of aluminium chloride compounding, China Environmental Science, 2008,28 (11):1014-1019) by microbial flocculant and chlorine
Change calcium, aluminium polychloride compounding use, flocculating effect is greatly improved, but the aluminium polychloride dosage for using still compares
Greatly.
The content of the invention
It is an object of the invention to provide the microbial flocculant that a Bacillus species are produced, and it is micro- to further provide for this
Application of the biological flocculant in metal ion waste water is processed.
Realize that the technical scheme of the object of the invention is as follows：
The microbial flocculant that one Bacillus species are produced, is CCTCC NO by deposit number：The class bud of M 2016615
Spore bacillus Paenibacillus sp.LX03 are produced.
Described series bacillus Paenibacillus sp.LX03 are preserved in Chinese Typical Representative on November 3rd, 2016
Culture collection (CCTCC), preservation address is Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University collection.
The present invention also provides the preparation method of the microbial flocculant that above-mentioned series bacillus are produced, and comprises the following steps that：
Series bacillus Paenibacillus sp.LX03 are seeded in fermentation medium, cultivation temperature is 30 DEG C, and shaking table cultivates 2
～3 days, Jing after centrifugation removes thalline, the ethanol for adding 2～3 times of volumes separated out extracellular products to zymotic fluid, after extracellular products drying
It is dissolved in water, obtains described microbial flocculant.
The composition of described fermentation medium is as follows：Starch 20g/L, sodium dihydrogen phosphate 1g/L, potassium nitrate 3g/L, calcium chloride
0.07g/L, magnesium chloride 0.2g/L, green vitriol 0.0125g/L, manganese sulfate 0.003g/L, zinc chloride 0.0075g/L,
Solvent is distilled water, and medium pH is 7.0～7.5.
The present invention also provides a kind of built-up flocculant, and described built-up flocculant is by above-mentioned microbial flocculant and containing gold
Category ion or polymeric aluminum chlorides solution composition.
Preferably, described microbial flocculant and the mass ratio of metal ion compound are 1:1～5.
Preferably, described metal ion is iron ion or aluminium ion.
Preferably, the concentration of described microbial flocculant is 4mg/L, and the concentration of metal ion solution is 20mg/L.
Preferably, described microbial flocculant and the mass ratio of aluminium polychloride are 1:0.5～3.5, more preferably 1:
1.5。
Preferably, the concentration of described microbial flocculant is 4mg/L, and the concentration of polymeric aluminum chlorides solution is 6mg/L.
Further, the microbial flocculant that the above-mentioned series bacillus of present invention offer are produced gives up in process metal ion
Application in water.
The present invention further provides application of the above-mentioned built-up flocculant in metal ion waste water is processed.
Compared with prior art, microbial flocculant of the invention is nontoxic, biodegradable, and metal ion or is polymerized
Liquor alumini chloridi is carried out after compounding use, forms Synergistic, can improve flocculating effect, and the use agent of flocculant is greatly reduced
Amount, flocculation rate is fast, and floccule body is big and closely knit, and sinking speed is fast, can be applied to the process field of metal ion waste water.
Description of the drawings
Fig. 1 is series bacillus Paenibacillus sp.LX03 colonial morphology figures.
Fig. 2 is microbial flocculant of the present invention in flocculation aid (2.7mM CaCl2·2H2O the flocculating effect figure in the presence of).
Fig. 3 is that microbial flocculant of the present invention processes the flocculating effect figure containing metal ion waste water.
Fig. 4 is the AlCl of microbial flocculant of the present invention and variable concentrations3·6H2O carries out the flocculating effect of compounding use
Figure.
Fig. 5 is the flocculation that microbial flocculant of the present invention and the aluminium polychloride (PAC) of variable concentrations carry out compounding use
Design sketch.
Specific embodiment
With reference to embodiment and accompanying drawing, the invention will be further described.
Embodiment 1
The screening of Paenibacillus sp.LX03 is separated and identified.
(1) screening and separation of bacterial strain
By the pedotheque gathered by Jiangsu Province's Yancheng City distillation water dissolves, supernatant dilution 10 is taken after 1h2～106Times,
Htm solid mediums are prepared, the sample of 100 μ L previously dilutions is respectively coated, in being placed in biochemical cultivation case, under conditions of 30 DEG C
2～3d of culture.The bacterial strain that colony characteristicses on culture dish have notable difference is selected, is equally purified using method of dilution butteron on plate,
Until obtaining single bacterial strain, its colonial morphology is as shown in Figure 1.
The composition of Htm fluid nutrient mediums is as follows：Starch 20g/L, sodium dihydrogen phosphate 1g/L, potassium nitrate 3g/L, calcium chloride
0.07g/L, magnesium chloride 0.2g/L, green vitriol 0.0125g/L, manganese sulfate 0.003g/L, zinc chloride 0.0075g/L,
Solvent is distilled water, and medium pH is 7.0～7.5.
(2) identification of bacterial strain
To screen the core DNA of the one plant of bacterial classification for obtaining as template, it is with the universal primer that the PCR of 16S rRNA genes is expanded
Primer, enters performing PCR amplification, determines its complete sequence.The 16S rDNA gene orders of bacterial strain are committed into GenBank databases
(the GenBank numbers of logging in are KX503263), with the sequence in GenBank databases online tetraploid rice is carried out, and is as a result shown,
The sequence similarity of LX03 and Paenibacillus alkaliterrae strain KSL-134 reaches 97%, with
Paenibacillus harenae strain B519 and Paenibacillus agarexedens strain DSM 1327
Sequence similarity reach 96%.Can determine that the bacterial classification belongs to bacillus genus, be named as Paenibacillus
sp.LX03。
Embodiment 2
The preparation technology of the microbial flocculant that Paenibacillus sp.LX03 bacterial strains are produced
Series bacillus Paenibacillus sp.LX03 are seeded to fermentation medium, 30 DEG C of cultivation temperature, and 250rpm shakes
Bed is cultivated 2～3 days, obtains the zymotic fluid containing the microbial flocculant, after 7000rpm centrifugation 10min remove thalline,
The ethanol that 2～3 times of volumes are added in zymotic fluid separates out extracellular products, and distillation water dissolves are added after extracellular products drying, obtains
To described microbial flocculant.Fermentation medium is above-mentioned Htm fluid nutrient mediums.
Embodiment 3
The flocculating property research of the microbial flocculant that Paenibacillus sp.LX03 bacterial strains are produced.
Aqueous suspension ofkaolin (4g/L) is prepared as the flocculating property of simulated wastewater test microbes flocculant.Concrete grammar
For：50mL aqueous suspension ofkaolins (4g/L) are placed in 50mL beakers, a certain amount of microbial flocculant are added, under normal temperature
100rpm magnetic agitations 1min, after static 5min, take the liquid sample of intermediate altitude, under spectrophotometric 550nm wavelength
Its absorbance is determined, microbial flocculant is replaced as a control group with distilled water.
Flocculating rate computing formula is：
Flocculating rate=(A ﹣ B)/A × 100%
Wherein, A is absorbance of the control group at 550nm, and B is absorbance of the experimental group at 550nm.
(1) flocculating properties of microbial flocculant
Calcium chloride is the conventional flocculation aid of microbial flocculant, using 2.7mM CaCl2·2H2O is and micro- as flocculation aid
Biological flocculant together adds aqueous suspension ofkaolin, determines the flocculating effect of variable concentrations microbial flocculant.As shown in Fig. 2
Flocculating rate reaches 83.2% when the concentration of microbial flocculant is 15mg/L.
(2) different metal ions and flocculating effect during microbial flocculant compounding use
Using Fe2(SO4)3, AlCl3·6H2O, ZnCl2, CaCl2·2H2O, MgSO4, MnSO4, KNO3, the solution such as NaCl
(final concentration of 20mg/L), with microbial flocculant (10mg/L) compounding use, microbial flocculant and metal ionization are carried out
The compound proportion (mass ratio) of compound is 1：2, microbial flocculant is determined with flocculation effect during different metal ions compounding use
Really.As shown in figure 3, and Fe3+And Al3+When carrying out compounding use, flocculating rate respectively reaches 92% and 97.3%.
(3) variable concentrations aluminium ion and flocculating effect during microbial flocculant compounding use
By the AlCl of final concentration of 4mg/L, 8mg/L, 12mg/L, 16mg/L, 20mg/L3·6H2O solution is wadded a quilt with cotton with microorganism
Solidifying agent (4mg/L) compounding use, microbial flocculant and AlCl3·6H2The compound proportion (mass ratio) of O is 1：1～5, determine not
With flocculating effect when aluminium ion concentration and microbial flocculant compounding use.As shown in figure 4, working as AlCl3·6H2O dosages
More than 8mg/L, compounding use is carried out with microbial flocculant, than using AlCl merely3·6H2The flocculating rate raising 10 of O～
20%, work as AlCl3·6H2When O is 20mg/L, microbial flocculant is 1 with the compound proportion of aluminium ion solution:5, flocculating rate is most
Height, reaches 97.6%.
(4) variable concentrations aluminium polychloride and flocculating effect during microbial flocculant compounding use
By final concentration of 2mg/L, 4mg/L, 6mg/L, 8mg/L, 12mg/L, 14mg/L aluminium polychloride (active ingredient
30%) with microbial flocculant (4mg/L), microbial flocculant is 1 with the compound proportion (mass ratio) of aluminium polychloride：0.5
～3.5, variable concentrations aluminium polychloride is determined with flocculating effect during microbial flocculant compounding use.As shown in figure 5, compounding
Flocculating rate when using is high more than the flocculating rate for using aluminium polychloride merely, particularly carries in aluminium polychloride low concentration
Rise larger, when the concentration of aluminium polychloride is 2mg/L, compounding use can bring up to 81% flocculating rate from 50.3%, and when poly-
When the concentration for closing aluminium chloride is 6mg/L, microbial flocculant is 1 with the compound proportion of aluminium polychloride:1.5, flocculating rate just reaches
To 94.3%, hereafter flocculating effect tends towards stability.
In sum, the microbial flocculant that series bacillus (Paenibacillus sp.) LX03 of the invention is produced,
During with iron ion, aluminium ion compounding use, with good flocculating effect, when processing containing iron ion, aluminum ions sewage,
The microbial flocculant effect is obvious, especially, when the iron ion, aluminium ion in sewage only has relatively low concentration, micro- life
Thing flocculant just can play a role.The microbial flocculant of the present invention processes dirty with inorganic flocculating agent aluminium polychloride compounding use
During water, flocculating effect is very notable, and the dosage of aluminium polychloride can be greatly reduced, particularly with low dosage polyaluminium
During aluminium compounding use, effect is lifted larger.The microbial flocculant that the present invention is provided is a kind of water process flocculation of good performance
Agent and flocculant aid, have potential using value in terms of the waste water such as life, industry are processed.
SEQUENCE LISTING
<110>Institutes Of Technology Of Nanjing
<120>Microbial flocculant and its application that one Bacillus species are produced
<130> 2016
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1518
<212> DNA
<213>Series bacillus（Paenibacillus sp.）
<400> 1
gagtttgatc ctggctcagg acgaacgctg gcggcgtgcc taatacatgc aagtcgagcg 60
gatcaagtat ttcccttggg aaatacgttg gttagcggcg gacgggtgag taacacgtag 120
gtaacctgcc tgtaagaccg ggataacatt cggaaacgaa tgctaatacc ggatacgcga 180
atgggtcgca tgacccgttc gggaaagacg gagcaatctg tcacttacag atggacctgc 240
ggcgcattag ctagttggtg aggtaacggc tcaccaaggc gacgatgcgt agccgacctg 300
agagggtgat cggccacact gggactgaga cacggcccag actcctacgg gaggcagcag 360
tagggaatct tccgcaatgg gcgaaagcct gacggagcaa cgccgcgtga gtgatgaagg 420
ttttcggatc gtaaagctct gttgccaggg aagaatgctt gggagagtaa ctgctcccaa 480
ggtgacggta cctgagaaga aagccccggc taactacgtg ccagcagccg cggtaatacg 540
tagggggcaa gcgttgtccg gaattattgg gcgtaaagcg cgcgcaggcg gctgattaag 600
tctggtgttt aaggctgggg ctcaaccccg gttcgcactg gaaactggtt ggcttgagtg 660
cagaagagga aagtggaatt ccacgtgtag cggtgaaatg cgtagagatg tggaggaaca 720
ccagtggcga aggcgacttt ctgggctgta actgacgctg aggcgcgaaa gcgtggggga 780
gcaaacagga ttagataccc tggtagtcca cgccgtaaac gatgaatgct aggtgttagg 840
ggtttcgata cccttggtgc cgaagttaac acattaagca ttccgcctgg ggagtacggt 900
cgcaagactg aaactcaaag gaattgacgg ggacccgcac aagcagtgga gtatgtggtt 960
taattcgaag caacgcgaag aaccttacca ggtcttgaca tccctctgac cgctctagag 1020
atagggcttc ccttcggggc agaggagaca ggtggtgcat ggttgtcgtc agctcgtgtc 1080
gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt gatcttagtt gccagcaggt 1140
aaggctgggc actctaggat gactgccggt gacaaaccgg aggaaggtgg ggatgacgtc 1200
aaatcatcat gccccttatg acctgggcta cacacgtact acaatggccg atacaacggg 1260
aagcgaagcc gcgaggcgga gccaatccta ccaaagtcgg tctcagttcg gattgcaggc 1320
tgcaactcgc ctgcatgaag tcggaattgc tagtaatcgc ggatcagcat gccgcggtga 1380
atacgttccc gggtcttgta cacaccgccc gtcacaccac gagagtttac aacacccgaa 1440
gccggtgggg taacccgcaa gggagctagc cgtcgaaggt ggggtagatg attggggtga 1500
agtcgtaaca aggtaacc 1518
Claims (10)
1. a Bacillus species produce microbial flocculant, it is characterised in that by deposit number be CCTCC NO：M
2016615 series bacillus Paenibacillus sp.LX03 are produced.
2. the preparation method of the microbial flocculant that series bacillus according to claim 1 are produced, it is characterised in that tool
Body step is as follows：Series bacillus Paenibacillus sp.LX03 are seeded in fermentation medium, cultivation temperature is 30
DEG C, shaking table is cultivated 2～3 days, and Jing after centrifugation removes thalline, the ethanol for adding 2～3 times of volumes separates out extracellular products, born of the same parents to zymotic fluid
It is dissolved in water after outer product drying, obtains described microbial flocculant.
3. the preparation method of the microbial flocculant that series bacillus according to claim 2 are produced, it is characterised in that institute
The composition of the fermentation medium stated is as follows：Starch 20g/L, sodium dihydrogen phosphate 1g/L, potassium nitrate 3g/L, calcium chloride 0.07g/L, chlorine
Change magnesium 0.2g/L, green vitriol 0.0125g/L, manganese sulfate 0.003g/L, zinc chloride 0.0075g/L, solvent is distillation
Water, medium pH is 7.0～7.5.
4. the built-up flocculant that microbial flocculant according to claim 1 is constituted, it is characterised in that by microbial flocculation
Agent is constituted with metal ion or polymeric aluminum chlorides solution.
5. built-up flocculant according to claim 4, it is characterised in that described microbial flocculant and metal ionization
The mass ratio of compound is 1:1～5.
6. built-up flocculant according to claim 4, it is characterised in that described metal ion be iron ion or aluminium from
Son.
7. built-up flocculant according to claim 4, it is characterised in that described microbial flocculant and aluminium polychloride
Mass ratio be 1:0.5～3.5.
8. built-up flocculant according to claim 7, it is characterised in that described microbial flocculant and aluminium polychloride
Mass ratio be 1:1.5.
9. application of the microbial flocculant according to claim 1 in metal ion waste water is processed.
10. the application according to the arbitrary described built-up flocculant of claim 4 to 8 in metal ion waste water is processed.
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| CN109574294A (en) * | 2018-11-12 | 2019-04-05 | 武汉华喻燃能工程技术有限公司 | A method of heavy metal ion is removed using pulullan polysaccharide and aluminium chloride |
| CN112553188A (en) * | 2020-10-27 | 2021-03-26 | 杭州秀川科技有限公司 | Carbon source slow-release wax loaded with microorganisms as well as preparation method and application thereof |
| CN114874941A (en) * | 2022-05-11 | 2022-08-09 | 淮阴师范学院 | Paenibacillus phyllophilus with raw starch hydrolysis capacity and application thereof |
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| 姜彬慧等: "类芽孢杆菌产絮凝多糖发酵条件优化及成分分析", 《环境科学研究》 * |
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| CN109574294A (en) * | 2018-11-12 | 2019-04-05 | 武汉华喻燃能工程技术有限公司 | A method of heavy metal ion is removed using pulullan polysaccharide and aluminium chloride |
| CN112553188A (en) * | 2020-10-27 | 2021-03-26 | 杭州秀川科技有限公司 | Carbon source slow-release wax loaded with microorganisms as well as preparation method and application thereof |
| CN114874941A (en) * | 2022-05-11 | 2022-08-09 | 淮阴师范学院 | Paenibacillus phyllophilus with raw starch hydrolysis capacity and application thereof |
| CN114874941B (en) * | 2022-05-11 | 2023-08-11 | 淮阴师范学院 | Paenibacillus phylloides with starch hydrolysis capability and application thereof |
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