CN109207464A - A kind of compound immobilization microorganism particles preparation and its application - Google Patents
A kind of compound immobilization microorganism particles preparation and its application Download PDFInfo
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- CN109207464A CN109207464A CN201710531184.8A CN201710531184A CN109207464A CN 109207464 A CN109207464 A CN 109207464A CN 201710531184 A CN201710531184 A CN 201710531184A CN 109207464 A CN109207464 A CN 109207464A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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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
- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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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
- C02F2101/30—Organic compounds
- C02F2101/308—Dyes; Colorants; Fluorescent 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
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
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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
- C02F2101/30—Organic compounds
- C02F2101/40—Organic compounds containing sulfur
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- Health & Medical Sciences (AREA)
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- General Health & Medical Sciences (AREA)
- Inorganic Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a kind of preparation methods of compound immobilization microorganism particles, belong to water treatment field.The technical solution of the present invention is to provide a kind of immobilization particles of capsule-type, it is not only easy to the growth of microorganism, and the immobilization particle of immobilization particle feature is had both, its specific method includes: (1) obtains efficient Photosynthetic bacterium strain by enrichment culture；(2) concentration of photosynthetic bacteria；(3) preparation of glass fibre ball；(4) photosynthetic bacteria Multiplying culture in glass fibre ball；(5) configuration of embedded material (polyvinyl alcohol, polyvinyl alcohol-sodium alginate, sodium alginate etc.)；(6) synthesis of compound Immobilized photosynthetic bacteria particle.Particle prepared by microorganism immobilization method of the present invention has stronger mechanical performance, and due to the porosity and translucency of glass fibre ball, the mass-transfer performance for overcoming traditional embedded particles is poor, the lower problem of light utilization efficiency.
Description
Technical field
The invention belongs to water-treatment technology fields, and in particular to a kind of preparation method of efficient fixed photosynthetic bacterium particle
And its application in water process.
Background technique
Photosynthetic bacteria (Photosynthetic bacteria, PSB) is one kind using light as the energy, can be in anaerobism illumination
Or photosynthesis is carried out as hydrogen donor and carbon source using organic matter, sulfide, the ammonia etc. in nature under aerobic dark condition
Microorganism.Since photosynthetic bacterial thallus is smaller, natural subsidence is difficult, there is thallus in practical applications and is lost and is separated by solid-liquid separation
Two problems.In order to solve both of these problems, it is necessary to constantly cultivate and add new fresh thalli, it is also necessary to be separated by solid-liquid separation
Processing work, thereby result in processing technological flow complexity, increase processing cost, seriously affected its popularization in production and answered
With.And immobilization technology can solve this problem, it is more that traditional process for fixation has investment and absorption method to apply, but with
Polyvinyl alcohol, sodium alginate, agar, gelatin etc. are the particle of embedding medium preparation, and bad mechanical property and embedded material will affect the
Five and product diffusivity, and then limit the application of this method, and absorption method makes bacterium mostly using porous material as carrier
It is attached to above, this process for fixation bacterium under hydraulic action is extremely easy to fall off from carrier.Therefore, the present invention utilizes
The advantages of investment and absorption method and the shortcomings that both overcoming is prepared for a kind of compound photosynthetic bacteria immobilization particle.
Summary of the invention
The purpose of the present invention is provide a kind of immobilized microorganism for main problem present in current bacteria adhension
Photosynthetic bacteria is adsorbed in glass fibre ball by technology by absorption method, then by investment in the outer surface of glass fibre ball
Thin film is coated, so that immobilization particle is made, the shortcomings that this immobilization particle not only overcomes traditional process for fixation,
And immobilization particle is significantly improved to the treatment effect of waste water.
The purpose of the present invention is what is be achieved through the following technical solutions:
The present invention provides a kind of compound photosynthetic bacteria immobilization filler (abbreviation immobilization particle), described compound photosynthetic
Bacteria adhension filler is prepared as follows:
(1) photosynthetic bacteria is cultivated under illumination anaerobic condition to logarithmic growth phase in photosynthetic bacteria liquid culture medium, Gu
Liquid separation discards supernatant liquid, collects photosynthetic bacterial thallus, then thallus is resuspended in sterile saline and is obtained with dry mycelium
After quality meter concentration is the photosynthetic bacteria concentrate of 1-6g/L, glass fibre ball is added, passes through the suction-operated of glass fibre ball
It is adsorbed on photosynthetic bacteria in the cellular structure of glass fibre ball, obtains the glass fibre ball of absorption photosynthetic bacteria；
(2) preparation of reagent is embedded: polyvinyl alcohol, sodium alginate, polyvinyl alcohol or sodium alginate is soluble in water, it prepares
At the embedding reagent of mass fraction 7%~15%；
(3) by the glass fibre ball table of step (2) embedding coated with agents obtained absorption photosynthetic bacteria obtained by step (1)
Face, be then totally submerged coated glass fibre ball keeps boric acid water-soluble in the boric acid aqueous solution of mass fraction 3%~6%
In liquid, at 4~18 DEG C crosslinking fix 12~for 24 hours, being in neutrality wash with distilled water to solution after taking-up can be obtained described answer
Mould assembly photosynthetic bacteria immobilization filler.
Further, photosynthetic bacteria liquid culture medium final concentration composition of the present invention are as follows: yeast extract 0.5g/L, peptone
0.5g/L、CH3COONa 3g/L、NH4Cl 0.1g/L、NaCl 0.5g/L、NaHCO3 0.5g/L、K2HPO4 0.2g/L、
MgSO4·7H2O 0.1g/L、CaCl20.1g/L, solvent are water, initial pH 7~8.
Further, glass fibre ball of the present invention the preparation method comprises the following steps: first glass fibre is sterilized, then by glass fibre
Mechanical heat ligation is carried out, is prepared into uniform sphere, cleaning, drying is to get the glass fibre ball.
Further, the mass ratio of photosynthetic bacteria concentrate of the present invention and glass fibre ball is 10~50:1.
Further, described be separated by solid-liquid separation under the revolving speed by 10000~12000rpm of step (1) of the present invention is centrifuged realization.
Further, photosynthetic bacteria of the present invention is Rhodopseudomonas (Rhodopseudomonas palustris) light
Close bacterium.
In addition, the answering in processing waste water from dyestuff the present invention also provides the compound photosynthetic bacteria immobilization filler
With.The dyestuff is methylene blue or reactive brilliant red.
The invention has the benefit that particle prepared by microorganism immobilization method of the present invention has stronger mechanicalness
Can, and due to the porosity and translucency of glass fibre ball, the mass-transfer performance for overcoming traditional embedded particles is poor, light utilization efficiency compared with
Low problem.
Detailed description of the invention
Fig. 1: glass fibre ball.
Fig. 2: compound Immobilized photosynthetic bacteria particle prepared by embodiment 1.
Fig. 3: decolorizing effect of the immobilization particle to dyestuff.In figure, abscissa E1 represents embodiment 1, and E2 represents embodiment
2, C1 represent comparative example 1, and C2 represents comparative example 2.
Specific embodiment
The present invention is described further combined with specific embodiments below, but protection scope of the present invention is not limited in
This.
The preferred Rhodopseudomonas of photosynthetic bacteria (Rhodopseudomonas palustris) photosynthetic bacteria of the present invention, light
It closes bacterium seed liquor and is purchased from Zhejiang Ding Long group.
For the sterile water that the present invention uses for the deionized water handled by high-pressure steam sterilizing pan, treatment conditions are temperature
It 121 DEG C, maintains 30 minutes under conditions of pressure 0.105Mpa.
Embodiment 1:
One, the preparation of glass fibre ball
It takes 3g glass fibre to sterilize in high-pressure steam sterilizing pan, then carries out the glass fibre ball of different packed densities
Mechanical heat ligation, is prepared into uniform glass fibre ball, cleans in sterile water, dry to get glass fibre ball.
Two, the preparation of photosynthetic bacteria concentrate
1. configuring photosynthetic bacteria liquid culture medium
Weigh yeast extract 0.5g, peptone 0.5g, CH3COONa 3g、NH4Cl 0.1g、NaCl 0.5g、NaHCO3
0.5g、K2HPO4 0.2g、MgSO4·7H2O 0.1g、CaCl20.1g is added in 1000mL distilled water, is with mass fraction
The HCI of 10% Na0H and 10% adjusts pH 7, stirs evenly, as photosynthetic bacteria liquid culture medium.
2. the enrichment culture of photosynthetic bacteria
200 milliliters of photosynthetic bacteria liquid culture mediums are prepared, pours into 250 milliliters of conical flasks and carries out high pressure steam sterilization,
In, sterilising temp is 115 DEG C, and sterilization time is 30 minutes, takes out conical flask and is cooled to room temperature.7 ring solids are pipetted with oese
In the photosynthetic bacteria to conical flask saved in culture medium, mixes, will be wrapped, be placed in incubator around taper bottleneck with gauze
Light intensity is illumination Anaerobic culturel 48h under the conditions of 2000lux, and cultivation temperature is 30 DEG C to get photosynthetic bacteria enrichment culture liquid.
3. the preparation of photosynthetic bacteria concentrate
Take photosynthetic bacteria enrichment culture liquid 20mL in centrifuge tube, revolving speed be 10000 revs/min under conditions of high speed from
The heart discards supernatant liquid, collects thallus, somatic cells are resuspended in sterile saline, obtains the light that concentration is 5g/L
Close bacteria concentrates.
Three, the preparation of photosynthetic bacteria immobilization particle
1. embedding the preparation of reagent
Polyvinyl alcohol is added to the water, heats, stir, dissolve, obtains embedding reagent；Wherein, the mass fraction of polyvinyl alcohol
It is 7%.
2. the configuration of immobilized reagent
Boric acid is added to the water, stirs, dissolve, obtain fixating reagent；Wherein, the mass fraction of immobilized reagent boric acid is
3%.
3. absorption of the glass fibre ball to bacterium
The glass fibre ball wash clean of 0.5g is weighed, is dried, is added to the photosynthetic bacteria that 100mL concentration is 5g/L and is inhaled
It is attached.
4. the preparation of immobilization particle
By the embedding coated with agents prepared in the glass fibre ball surface for being adsorbed with photosynthetic bacteria, then put it into
In immobilized reagent, 12h is fixed in 4 DEG C of refrigerators, takes out, with sterile water wash, that is, obtains compound Immobilized photosynthetic bacteria.
Four, the waste water index of methylene blue: 20mg/L.
Five, the Activity determination of immobilization particle
It takes 0.5g immobilization particle to be placed in the methylene blue waste water of 200mL, is 30 DEG C in temperature, intensity of illumination is
2000lux, under conditions of handle 72h.
Percent of decolourization 82% of the compound Immobilized photosynthetic bacteria to methylene blue known to Fig. 3 (E1).
Comparative example 1:
One, the preparation of photosynthetic bacteria concentrate
1. configuring photosynthetic bacteria liquid culture medium
Weigh yeast extract 0.5g, peptone 0.5g, CH3COONa 3g、NH4Cl 0.1g、NaCl 0.5g、NaHCO3
0.5g、K2HPO4 0.2g、MgSO4·7H2O 0.1g、CaCl20.1g is added in 1000mL distilled water, is with mass fraction
The HCI of 10% Na0H and 10% adjusts pH 7, stirs evenly, as photosynthetic bacteria liquid culture medium.
2. the enrichment culture of photosynthetic bacteria
200 milliliters of photosynthetic bacteria liquid culture mediums are prepared, pours into 250 milliliters of conical flasks and carries out high pressure steam sterilization,
In, sterilising temp is 115 DEG C, and sterilization time is 30 minutes, takes out conical flask and is cooled to room temperature.7 ring solids are pipetted with oese
In the photosynthetic bacteria to conical flask saved in culture medium, mixes, will be wrapped, be placed in incubator around taper bottleneck with gauze
Light intensity is illumination Anaerobic culturel 48h under conditions of 2000lux, and cultivation temperature is 30 DEG C to get photosynthetic bacteria enrichment culture liquid.
3. the preparation of photosynthetic bacteria concentrate
Take photosynthetic bacteria enrichment culture liquid 20mL in centrifuge tube, revolving speed be 10000 revs/min under conditions of high speed from
The heart discards supernatant liquid, collects thallus, somatic cells are resuspended in sterile saline, obtains the light that concentration is 5g/L
Close bacteria concentrates.
Two, the preparation of photosynthetic bacteria immobilization particle
1. embedding the preparation of reagent
Polyvinyl alcohol is added to the water, heats, stir, dissolve, obtains embedding reagent；Wherein, the mass fraction of polyvinyl alcohol
It is 7%.
2. the configuration of immobilized reagent
Boric acid is added to the water, stirs, dissolve, obtain fixating reagent；Wherein, the mass fraction of immobilized reagent boric acid is
3%.
3. the preparation of immobilization particle
Bacteria concentrates are placed in embedding reagent, are stirred evenly, are instilled in immobilized reagent with syringe, in 4 DEG C of ice
8-12h is fixed in case, is taken out, with sterile water wash, that is, is obtained compound Immobilized photosynthetic bacteria.
Three, the waste water index of methylene blue: 20mg/L.
Four, the Activity determination of immobilization particle
It takes 0.5g immobilization particle to be placed in the methylene blue waste water of 200mL, is 30 DEG C in temperature, intensity of illumination is
2000lux, under conditions of handle 72h.
Immobilized photosynthetic bacteria is to the percent of decolourization of methylene blue up to 53% known to Fig. 3 (C1), hence it is evident that lower than compound
Percent of decolourization of the immobilization particle to methylene blue.
Embodiment 2:
One, the preparation of glass fibre ball: consistent with embodiment 1
Two, the preparation of photosynthetic bacteria concentrate: consistent with embodiment 1
Three, the preparation of photosynthetic bacteria immobilization particle: consistent with embodiment 1
Four, reactive brilliant red waste strength index: 100mg/L.
Five, the Activity determination of immobilization particle
It takes 0.5g immobilization particle to be placed in the reactive brilliant red waste water of 200mL, is 30 DEG C in temperature, intensity of illumination is
2000lux, under conditions of handle 72h.
Compound Immobilized photosynthetic bacteria is to the percent of decolourization of reactive brilliant red up to 91% known to Fig. 3 (E2).
Comparative example 2:
One, the preparation of glass fibre ball: consistent with comparative example 1
Two, the preparation of photosynthetic bacteria concentrate: consistent with comparative example 1
Three, the preparation of photosynthetic bacteria immobilization particle: consistent with comparative example 1
Four, reactive brilliant red waste strength index: 100mg/L.
Five, the Activity determination of immobilization particle
It takes 0.5g immobilization particle to be placed in the reactive brilliant red waste water of 200mL, is 30 DEG C in temperature, intensity of illumination is
2000lux, under conditions of handle 72h.
Compound Immobilized photosynthetic bacteria is to the percent of decolourization of reactive brilliant red up to 75% known to Fig. 3 (C2).
Embodiment 3:
One, the preparation of glass fibre ball: consistent with embodiment 1
Two, the preparation of photosynthetic bacteria concentrate: consistent with embodiment 1
Three, the preparation of photosynthetic bacteria immobilization particle
3. embedding the preparation of reagent
Sodium alginate is added to the water, heats, stir, dissolve, obtains embedding reagent；Wherein, the mass fraction of sodium alginate
It is 15%.
4. the configuration of immobilized reagent
Boric acid is added to the water, stirs, dissolve, obtain fixating reagent；Wherein, the mass fraction of immobilized reagent boric acid is
6%.
3. absorption of the glass fibre ball to bacterium
The glass fibre ball wash clean of 0.5g is weighed, is dried, is added to the photosynthetic bacteria that 100mL concentration is 1g/L and is inhaled
It is attached.
4. the preparation of immobilization particle
By the embedding coated with agents prepared in the glass fibre ball surface for being adsorbed with photosynthetic bacteria, then put it into
In immobilized reagent, 12h is fixed in 4 DEG C of refrigerators, takes out, with sterile water wash, that is, obtains compound Immobilized photosynthetic bacteria.
Four, reactive brilliant red waste strength index: 100mg/L.
Four, reactive brilliant red waste strength index: 100mg/L.
Five, the Activity determination of immobilization particle
It takes 0.5g immobilization particle to be placed in the reactive brilliant red waste water of 200mL, is 30 DEG C in temperature, intensity of illumination is
2000lux, under conditions of handle 72h.
Compound Immobilized photosynthetic bacteria is to the percent of decolourization of reactive brilliant red up to 83%.
Claims (8)
1. a kind of compound photosynthetic bacteria immobilization filler, it is characterised in that the compound photosynthetic bacteria immobilization filler press with
The preparation of lower section method:
(1) by photosynthetic bacteria, culture to logarithmic growth phase, solid-liquid divides under illumination anaerobic condition in photosynthetic bacteria liquid culture medium
Supernatant is abandoned, collects photosynthetic bacterial thallus, then thallus is resuspended in sterile saline and is obtained with dry mycelium quality
After counting the photosynthetic bacteria concentrate that concentration is 1-6g/L, glass fibre ball is added, light is made by the suction-operated of glass fibre ball
Bacterial adsorption is closed in the cellular structure of glass fibre ball, obtains the glass fibre ball of absorption photosynthetic bacteria；
(2) preparation of reagent is embedded: polyvinyl alcohol, sodium alginate, polyvinyl alcohol or sodium alginate is soluble in water, it is configured to matter
Measure the embedding reagent of score 7%~15%；
(3) step (2) embedding coated with agents obtained is adsorbed to the glass fibre ball surface of photosynthetic bacteria in step (1) gained,
Then coated glass fibre ball is totally submerged makes boric acid aqueous solution in the boric acid aqueous solution of mass fraction 3%~6%
In, at 4~18 DEG C crosslinking fix 12~be in neutrality wash with distilled water to solution for 24 hours, after taking-up can be obtained it is described compound
Type photosynthetic bacteria immobilization filler.
2. compound photosynthetic bacteria immobilization filler as described in claim 1, it is characterised in that: photosynthetic bacteria is red false unit cell
Pseudomonas (Rhodopseudomonas palustris) photosynthetic bacteria.
3. compound photosynthetic bacteria immobilization filler as described in claim 1, it is characterised in that: the photosynthetic bacteria liquid training
Support base final concentration composition are as follows: yeast extract 0.5g/L, peptone 0.5g/L, CH3COONa 3g/L、NH4Cl 0.1g/L、NaCl
0.5g/L、NaHCO3 0.5g/L、K2HPO4 0.2g/L、MgSO4·7H2O 0.1g/L、CaCl20.1g/L, solvent are water, just
Beginning pH 7~8.
4. compound photosynthetic bacteria immobilization filler as described in claim 1, it is characterised in that the system of the glass fibre ball
Preparation Method are as follows: first glass fibre sterilizes, then glass fibre is subjected to mechanical heat ligation, is prepared into uniform sphere, clearly
Drying is washed to get the glass fibre ball.
5. compound photosynthetic bacteria immobilization filler as described in claim 1, it is characterised in that: the photosynthetic bacteria concentrate
Mass ratio with glass fibre ball is 10~50:1.
6. compound photosynthetic bacteria immobilization filler as described in claim 1, it is characterised in that: step (1) solid-liquid point
It is realized from by being centrifuged under the revolving speed of 10000~12000rpm.
7. application of the compound photosynthetic bacteria immobilization filler as described in claim 1 in processing waste water from dyestuff.
8. it is the use as claimed in claim 7, it is characterized in that: the dyestuff is methylene blue or reactive brilliant red.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110195052A (en) * | 2019-05-31 | 2019-09-03 | 浙江工业大学 | A kind of photosynthetic bacteria immobilization particle and the preparation method and application thereof |
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| CN104743677A (en) * | 2015-03-06 | 2015-07-01 | 浙江工业大学 | Method for treating wastewater by utilizing immobilized photosynthetic bacterium coupled film reaction system |
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2017
- 2017-07-03 CN CN201710531184.8A patent/CN109207464A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104743677A (en) * | 2015-03-06 | 2015-07-01 | 浙江工业大学 | Method for treating wastewater by utilizing immobilized photosynthetic bacterium coupled film reaction system |
Non-Patent Citations (3)
| Title |
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| ANATOLY A.TSYGANKOV 等: "Photobioreactor with photosynthetic bacteria immobilized on porous glass for hydrogen photoproduction", 《JOURNAL OF FERMENTATION AND BIOENGINEERING》 * |
| 许志强: "固定化光合细菌在暗纹东方鲀养殖中的应用", 《中国优秀博硕士学位论文全文数据库(硕士)农业科技辑》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110195052A (en) * | 2019-05-31 | 2019-09-03 | 浙江工业大学 | A kind of photosynthetic bacteria immobilization particle and the preparation method and application thereof |
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