CN106636057A - Nutrient source immobilized sulfate reducing bacterium globules as well as preparation method and application thereof - Google Patents

Nutrient source immobilized sulfate reducing bacterium globules as well as preparation method and application thereof Download PDF

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Publication number
CN106636057A
CN106636057A CN201611081224.5A CN201611081224A CN106636057A CN 106636057 A CN106636057 A CN 106636057A CN 201611081224 A CN201611081224 A CN 201611081224A CN 106636057 A CN106636057 A CN 106636057A
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nutrient source
sulfate reducing
bead
globules
reducing bacteria
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CN201611081224.5A
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Chinese (zh)
Inventor
代丽花
李欣
张长
曾光明
刘云国
吴幼娥
田亚灵
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Hunan University
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Hunan University
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    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/04Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/10Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/14Enzymes or microbial cells immobilised on or in an inorganic carrier
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/10Inorganic compounds
    • C02F2101/20Heavy metals or heavy metal compounds
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/10Inorganic compounds
    • C02F2101/20Heavy metals or heavy metal compounds
    • C02F2101/22Chromium or chromium compounds, e.g. chromates

Abstract

The invention relates to a preparation method and application of nutrient source immobilized sulfate reducing bacterium globules. Immobilized globules take polyvinyl alcohol and sodium alginate as carriers, a saturated boric acid calcium chloride solution as a cross-linking agent and sodium lactate as a nutrient source. The preparation method comprises the specific steps: firstly, dissolving the polyvinyl alcohol and the sodium alginate in a water bath at a constant temperature; then, adding active carbon, calcium carbonate, silicon dioxide and free bacterium liquid; sucking a mixed solution by an injector and dropwise adding the mixed solution into the saturated boric acid calcium chloride solution; crossly linking for 24h to form the globules; washing the globules with sterile physiological saline and putting the globules into a sodium lactate solution for 24h; after washing, freezing and drying to obtain the nutrient source immobilized sulfate reducing bacterium globules. In the application of the nutrient source immobilized sulfate reducing bacterium globules prepared by the invention, the nutrient source immobilized sulfate reducing bacterium globules have a good removing effect on four types of heavy metal including Cu, Zn, Pb and Cd in sediment of Xiang River; the treatment is stable and the leaching rate is low. The material is convenient to separate, can be repeatedly utilized and does not cause secondary pollution; the nutrient source immobilized sulfate reducing bacterium globules has good economic benefits and a wide application prospect.

Description

A kind of nutrient source fixes sulfate reducing bacteria bead and preparation method thereof and purposes
Technical field
The invention belongs to heavy metal pollution processing technology field, and in particular to one kind is using in immobilization sulfate reducing bacteria The deposit of the heavy metal species of bead stabilization processes copper zinc lead chromium four pollution made by poly- nutrient source, and the repeatable profit of bead The property used.
Background technology
Life of the Heavy Metal Pollution in Sediment to the ambient environment ecosystem and people has huge harm.With close The industrialization and urbanization process of collection, these heavy metals Jing food chain into aquatic environment after can not be biodegradable, in biology Cylinder accumulation, destroys its metabolic activity, is detrimental to health and ecosystem balance.And the molecule in deposit affects The migration of heavy metal forms stable compound so that metal longer-term persistence, and the big pollution of toxicity is increasingly severe.Therefore one is sought Plant thorough and the technology of effective process Heavy Metal Pollution in Sediments is most important.The method of process heavy metal pollution traditional at present There are ion exchange, chemical precipitation, electrochemical process, absorption and biological treatment etc..But these method processing costs are high, can produce Secondary pollution, to subsequent treatment difficulty is increased.And microorganism treatment, low cost, efficient and non-secondary pollution is processed, it is to plant Environmentally friendly processing method.
Sulfate reducing bacteria, a kind of anaerobe can be by SO4 2-It is reduced to S2-, further give birth to metal ion reaction Into settlement thing.But especially bacterium is in free state to the metal concentration of high concentration meeting inhibiting sulfate reduction bacterium activity Under be restricted this biogenic sediment process.Therefore immobilization technology can effectively solve this problem.Polyvinyl alcohol and alginic acid Sodium is conventional carrier, and bead made by PVA-sodium boric methods has been widely used in the process of metallic pollution.Profit After will be microbial immobilized with immobilized microorganism technique, the ability to bear and degradation capability of toxic heavy metal be carried It is high.The selection of carbon source has important function for Growth of Sulfate Reducing Bacteria, and sodium lactate, can be in bead used as cohesion nutrient source Being made full use of by sulfate reducing bacteria can avoid organic contamination.By nutrient source fixation of bacteria, treatment effeciency is not only improved, may be used also Secondary pollution is avoided, while additionally aiding the separation and recycling of immobilized spherule particle.
The content of the invention
The technical problem to be solved is high cost, the low problem for the treatment of effeciency existed for prior art, Exploitation is a kind of efficiently, the repeatable immobilized spherule particle for processing Heavy Metal Pollution in Sediment for utilizing.
A kind of nutrient source proposed by the present invention fixes the preparation method of sulfate reducing bacteria bead, is by sulfate reducing bacteria It is embedded in the bead that polyvinyl alcohol, sodium alginate and calcium chloride are cross-linked to form, and is immersed in nutrient source, is prepared Immobilized spherule can make full use of nutrient source, it is to avoid produce organic contamination, comprise the following steps that:
(1) the sulfate reduction bacterial classification of purchase is carried out into enrichment culture, the composition of fluid nutrient medium is:The anhydrous sulphur of 0.5g/L Sour sodium, 1.0g/L ammonium chlorides, 0.1g/L calcium chloride, 2g/L magnesium sulfate, 0.5g/L dipotassium hydrogen phosphates, 3.5g/L sodium lactates, 1.0g/ L yeast extracts, 0.5g/L iron ammonium sulfates, ascorbic acid 0.5g/L, seal culture 7 days, like this step under the conditions of 37 DEG C of anaerobism Carry out continuous Amplification Culture;
(2) 9g polyvinyl alcohol and 3g sodium alginates are mixed to join in the deionized water of 100mL, are placed in constant temperature magnetic force and stir Mix on device stirring and heating water bath obtains mixed liquor A until being completely dissolved to 90 DEG C.
(3) 3g activated carbons, 3g silica, 0.5g calcium carbonate are slowly added to when stirring is cooled to room temperature to A mixed liquors, And be stirred continuously to be allowed to be well mixed and obtain mixed liquid B;By mixed liquid B, glass bar, syringe, saturation boric acid calcium chloride solution It is placed in sterilizing a period of time on ultraviolet sterilization platform.
(4) turn off uviol lamp, the sulfate reducing bacteria of enrichment culture is taken into 20mL and is added to stirring in sterilized solution B Mixed liquor C is uniformly obtained, then mixed liquor C is drawn with syringe and is added dropwise in 2% saturation boric acid calcium chloride solution, in room temperature Lower stirring solidification 24h;
(5) bead for having solidified is cleaned with SPSS and removes after boric acid that to be placed in aseptic 6g sodium lactates molten several times 24h in constant incubator is placed in liquid, during 4 DEG C of refrigerators are stored in after then being cleaned up with SPSS again;To make Bead freeze-drying and dehydrating after preserve.
In above-mentioned preparation method, every operation is aseptically carried out.
The nutrient source immobilization sulfate reducing bacteria bead prepared using the inventive method can be processed in deposit Tetra- kinds of metals of Cu, Zn, Pb, Cd.
Compared with prior art, it is an advantage of the current invention that:
1. the chemicals polyvinyl alcohol for using and sodium alginate, are common fixation supports, activated carbon specific surface area Greatly, the material price for producing bead is cheap, low cost.
2. the immobilized spherule preparation technology and simple to operate for making, prepares quick, repeats and utilizes, and is easy to reclaim, It is easy to
Realize industrialized production.
3. the addition of sodium lactate can make full use of by sulfate reducing bacteria, it is to avoid organic contamination.
4. product is nontoxic, environmentally friendly.
5. the clearance of the immobilized spherule heavy metal of the present invention is high, and Leaching is low, and stability is high.
Description of the drawings
Fig. 1 is that the ESEM of obtained nutrient source immobilization sulfate reducing bacteria bead in the embodiment of the present invention 1 is illustrated Figure.
Fig. 2 is the energy spectrum analysis figure that nutrient source fixes sulfate reducing bacteria bead after the embodiment of the present invention 1.
Fig. 3 is the fractions distribution figure of four heavy metal species in the embodiment of the present invention 1.
Fig. 4 is the Leaching design sketch of four kinds of metals in the embodiment of the present invention 1.
Specific embodiment
The present invention is described in further details below with reference to Figure of description and specific embodiment.
Embodiment 1:
A kind of nutrient source of embodiment of the present invention of the present invention 1 fixes sulfate reducing bacteria bead and processes Xiang River deposition Tetra- kinds of metals of Cu, Zn, Pb, Cd in thing, comprise the following steps:
A, by collection Jing 100 mesh sieve filter after Xiangjiang River sediments sample and immobilized spherule with 5:1 ratio adds Enter in the serum bottle for containing bacteria culture media to 150mL and vibrate 5 with 150rpm under 37 DEG C of environmental conditions in constant incubator After it, bead is collected by filtration, suspension is centrifuged 20min with 3000rpm, collects sediment sample and freeze-drying is used for BCR It is continuous to extract;
B, take cryodesiccated sample 0.2g after above-mentioned centrifugation and be placed in the polytetrafluoroethylene (PTFE) centrifuge tube of 50mL, add 20mL0.11mol/L acetic acid constant temperature oscillation 16h, add 10mL deionized waters and 20min are centrifuged in 3000r/min, separate supernatant Liquid is diluted to scale and shakes up in the volumetric flask of 50mL, uses flame atomic absorption spectrophotometer detectable concentration;
C, the hydroxylamine hydrochloride of 20mL0.1mol/L is added in b in residue, constant temperature oscillation 16h adds 10mL deionizations Water is centrifuged 20min in 3000r/min, and separation supernatant is diluted to scale and shakes up, inhaled with NITRATE BY FLAME ATOMIC in the volumetric flask of 50mL Receive spectrophotometer detectable concentration;
D, in c 10mL8.8mol/L hydrogen peroxide is added, stand 1h under room temperature, heating water bath is treated to 85 DEG C of digestion 1h Steam to closely dry and add 6mL8.8mol/L hydrogen peroxide, repetition aforesaid operations are cooled to after room temperature and add 20mL1mol/L vinegar Sour ammonium constant temperature oscillation 16h, adds 10mL deionized waters and 20min is centrifuged in 3000r/min, separates supernatant in the capacity of 50mL Scale is diluted in bottle to shake up, use flame atomic absorption spectrophotometer detectable concentration;
E, 10mLHCl is added toward residue in d, be heated to removing cooling after sample is remaining a small amount of at 100 DEG C, addition 5mLHNO3, 5mLHF, 3mLHClO4Cool down after 180 DEG C of intensification 1h, continue to be heated to 200 DEG C, emit to removing after a small amount of after white cigarette Cooling, liquid is transparent, pours out and is diluted to scale into the volumetric flask of 50mL and shakes up, and is examined with flame atomic absorption spectrophotometer Survey concentration.
It is in black that nutrient source obtained above fixes sulfate reducing bacteria bead outward appearance, is placed under ESEM and sees Examine, its surface texture is as shown in Figure 1, it can be seen that bead outside closely knit (a), heavy metal toxicity infringement can be resisted, be internally formed Network structure (b), there is provided easily matter transportation passage, zoogloea is attached to above, there is substantial amounts of rod bacterium (c).To bead Energy spectrum analysis is done, as shown in Figure 2, it can be seen that process outer surface after sample with bead and contain Cu, Zn, Pb, Cd, S, can speculate anti- Should after generate stable sulfide.
Fig. 3 is the various fractions distribution feelings that immobilized spherule processes Cu, Zn, Pb, Cd after sediment sample in the present embodiment Condition.As seen from the figure, compared with sample background values, four kinds of metals are big with content that sour solvent form is present after immobilized spherule process Big to reduce, oxidable state content is significantly improved, as a result show immobilized spherule use so that in deposit four heavy metal species with More stabilized form is present, and environmental hazard is reduced.
Fig. 4 is the steadiness that immobilized spherule processes effect after sediment sample in the present embodiment.It is computed, Cu, The clearance of tetra- kinds of metals of Zn, Pb, Cd is respectively 76.3%, 95.6%, 100% and 91.2%.Treatment effect is stablized, and leaches dense Degree is relatively low.

Claims (9)

1. a kind of preparation method and purposes of immobilization sulfate reducing bacteria bead, it is characterised in that the immobilized spherule shape Into cellular structure, zoogloea is supported on hole wall.
2. a kind of nutrient source as claimed in claim 1 fixes the preparation method of sulfate reducing bacteria bead, comprises the following steps:
(1) polyvinyl alcohol and sodium alginate are added in the deionized water of 100mL and are stirred, then water bath with thermostatic control is heated to 90 degree And 30min is kept, it is stirred continuously to being completely dissolved.
(2) by activated carbon, silica, calcium carbonate slowly adds in above-mentioned solution and is constantly stirred until homogeneous mixing, waits to mix Conjunction liquid is cooled to after room temperature and is placed in ultraviolet sterilizing a period of time, subsequently adds in mixed liquor bacterial suspension and constantly stirs Mix.
(3) during the mixed liquor syringe containing bacterial suspension dropwise to be instilled 2% saturation boric acid calcium chloride solution, in 24h is crosslinked under the conditions of being stirred at room temperature.Then it is immersed in sodium lactate solution, is kept 24h.
(4) subsequently bead saturated nacl aqueous solution is cleaned up to neutrality, freeze-drying obtains nutrient source immobilization sulfuric acid Salt reducing bacteria bead.Bead is stored in 4 DEG C of environment.
3. the preparation method of nutrient source immobilization sulfate reducing bacteria bead according to claim 2, it is characterised in that institute In stating the preparation of immobilized spherule, with polyvinyl alcohol and sodium alginate as carrier, saturation boric acid calcium chloride solution is used as crosslinking Agent, sodium lactate is cohesion nutrient source so that sulfate reducing bacteria is successfully embedded in inside bead, is conducive to the abundant of nutrient source Utilize, prevent secondary pollution.
4. a kind of nutrient source as claimed in claim 2 fixes the preparation method of sulfate reducing bacteria bead, it is characterised in that institute In stating the preparation of immobilized spherule, the mass fraction point of polyvinyl alcohol, sodium alginate, saturation boric acid calcium chloride and free bacterium solution Wei 9%, 3%, 2% and 20mL.
5. a kind of nutrient source as claimed in claim 2 fixes the preparation method of sulfate reducing bacteria bead, it is characterised in that institute In stating the preparation of immobilized spherule, activated carbon is added, specific surface area is big, increased adsorption area, and then is conducive to nutrient source Absorption.
6. a kind of nutrient source as claimed in claim 2 fixes the preparation method of sulfate reducing bacteria bead, it is characterised in that institute In stating the preparation of nutrient source immobilized spherule, bead can be placed in constant incubator, keep the activity of sulfate reducing bacteria.
7. a kind of nutrient source as claimed in claim 2 fixes the purposes of sulfate reducing bacteria bead, it is characterised in that with described The heavy metal species of Cu, Zn, Pb, Cd tetra- that immobilized spherule comes in stabilization processes deposit.
8. a kind of nutrient source as claimed in claim 2 fixes the purposes of sulfate reducing bacteria bead, it is characterised in that with described Immobilized spherule can make tetra- kinds of metals of Cu, Zn, Pb, Cd form stable sulfide, and leaching rate is low.
9. the purposes according to claim 7 and 8, it is characterised in that the ratio 1 of the immobilized spherule and deposit consumption: 5。
CN201611081224.5A 2016-11-30 2016-11-30 Nutrient source immobilized sulfate reducing bacterium globules as well as preparation method and application thereof Pending CN106636057A (en)

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CN109136215A (en) * 2018-08-22 2019-01-04 广州大学 A kind of preparation method and applications of immobilization sulfate reducing bacteria particle
CN109912044A (en) * 2019-03-19 2019-06-21 中国地质大学(北京) A kind of Waste water treatment medicament and its application method
CN109928516A (en) * 2019-04-20 2019-06-25 无锡吉致电子科技有限公司 A kind of efficient application for soaking cleanser and its depollution of environment
CN109928517A (en) * 2019-04-20 2019-06-25 无锡吉致电子科技有限公司 A kind of various dimensions water purifying device and its application
CN110452851A (en) * 2019-08-26 2019-11-15 天津科技大学 It is a kind of screening efficient degradation phenol, pyridine, quinoline flora method and application
CN110452900A (en) * 2019-08-26 2019-11-15 天津科技大学 A kind of preparation method for the compound fixation support of PVA-SA embedding degrading microorganism
CN111517477A (en) * 2020-05-14 2020-08-11 江西省科学院 Sulfate reducing bacteria embedded particles for treating ionic rare earth mine wastewater, and preparation method and application thereof

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109136215A (en) * 2018-08-22 2019-01-04 广州大学 A kind of preparation method and applications of immobilization sulfate reducing bacteria particle
CN109912044A (en) * 2019-03-19 2019-06-21 中国地质大学(北京) A kind of Waste water treatment medicament and its application method
CN109928516A (en) * 2019-04-20 2019-06-25 无锡吉致电子科技有限公司 A kind of efficient application for soaking cleanser and its depollution of environment
CN109928517A (en) * 2019-04-20 2019-06-25 无锡吉致电子科技有限公司 A kind of various dimensions water purifying device and its application
CN110452851A (en) * 2019-08-26 2019-11-15 天津科技大学 It is a kind of screening efficient degradation phenol, pyridine, quinoline flora method and application
CN110452900A (en) * 2019-08-26 2019-11-15 天津科技大学 A kind of preparation method for the compound fixation support of PVA-SA embedding degrading microorganism
CN111517477A (en) * 2020-05-14 2020-08-11 江西省科学院 Sulfate reducing bacteria embedded particles for treating ionic rare earth mine wastewater, and preparation method and application thereof
CN111517477B (en) * 2020-05-14 2021-01-22 江西省科学院 Sulfate reducing bacteria embedded particles for treating ionic rare earth mine wastewater, and preparation method and application thereof

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Application publication date: 20170510