CN105779427A - Preparation method of rhodococcus BH4-sodium alginate porous micro-spheres and application thereof - Google Patents
Preparation method of rhodococcus BH4-sodium alginate porous micro-spheres and application thereof Download PDFInfo
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- CN105779427A CN105779427A CN201610120977.6A CN201610120977A CN105779427A CN 105779427 A CN105779427 A CN 105779427A CN 201610120977 A CN201610120977 A CN 201610120977A CN 105779427 A CN105779427 A CN 105779427A
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- rhod
- sodium alginate
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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
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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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
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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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention discloses a preparation method of rhodococcus BH4-sodium alginate porous micro-spheres.The preparation method includes the following steps that separated rhodococcus BH4 is inoculated to an LB liquid culture medium for culture; the cultured rhodococcus BH4 is subjected to centrifugal treatment, and a rhodococcus BH4 suspension is prepared after centrifugation; the BH4 suspension and a sodium alginate suspension are mixed to obtain a BH4-sodium alginate suspension; a calcium chloride solution is added to the BH4-sodium alginate suspension to obtain the BH4-sodium alginate porous micro-spheres; finally cleaning, drying and soaking are performed.The invention further discloses application of the rhodococcus BH4-sodium alginate porous micro-spheres prepared by means of the preparation method on the aspect of MBR membrane pollution inhabitation.The rhodococcus BH4-sodium alginate porous micro-spheres prepared by means of the preparation method have high encapsulation rate, also perform biological membrane stripping on the two aspects of physics and biology and greatly improve the biological membrane stripping efficiency.
Description
Technical field
The present invention relates to the preparation method of a kind of Rhod BH4-sodium alginate porous microsphere, further relate to above-mentioned preparation side
The Rhod BH4-sodium alginate porous microsphere that method prepares application in terms of suppression MBR fouling membrane, belongs at sewage
Reason technical field.
Background technology
In more than 20 year of past, membrane technology quickly grows, and has become as a kind of new technique that wastewater efficient processes.Film
Bioreactor (MBR) is the combination of membrane separation technique and traditional active sludge, compared with traditional activated sludge process, and tool
Have that sludge yield is low, volumetric loading is high, organic removal rate is high, floor space is little and is prone to the advantages such as Automated condtrol,
But MBR membrane pollution problem (biomembrane on the microorganism flco+film surface of formation of deposits) remains obstruction membrane separation technique
One difficult problem of development, has had a strong impact on stability and the economy of this technique, has limited it and be widely used.
In terms of the domestic and international research about alleviation MBR membrane pollution problem is mainly around following 3: membrane material and membrane module
Structure optimization, adjustment reactor service condition, raising mud mixed liquid filtrability.Although these traditional measures achieve
Certain effect, but being formed of suppression film surface biological film is always the greatest problem that traditional method faces.
In recent years, the concept of bacterial community sensing cancellation is applied to control on membrane pollution problem, based on quorum sensing former
Reason, is added to AHL-QS digestive enzyme in MBR signaling molecule of degrading, thus suppresses MBR film surface biological film
Formed, but in the method, digestive enzyme easily decomposes, and easily runs off, so practice poor effect.Therefore there is this
Problem, therefore combines slow release method and is embedded in sodium alginate by strain, it is possible to effectively prevent loss and the inactivation of strain,
Strain plays and biofilm formation is risen inhibitory action.
Summary of the invention
Goal of the invention: it is micro-that the technical problem to be solved is to provide a kind of Rhod BH4-sodium alginate porous
The preparation method of ball.
The present invention also to solve the technical problem that the Rhod BH4-sodium alginate being to provide above-mentioned preparation method prepared is many
The application in terms of suppression MBR fouling membrane of the hole microsphere.
Summary of the invention: for solving above-mentioned technical problem, the technical solution adopted in the present invention is:
The preparation method of a kind of Rhod BH4-sodium alginate porous microsphere, specifically includes following operating procedure:
Step 1, is inoculated into Rhod BH4 isolated in MBR reaction tank on LB fluid medium, Yu Heng
Enrichment culture a period of time under the conditions of temperature；
Step 2, is centrifuged the Rhod BH4 after enrichment culture processing, is configured to Rhod BH4 after being centrifuged
Suspension；
Step 3, is mixed to get Rhod BH4-by the Rhod BH4 suspension of step 2 and sodium alginate suspension
Sodium alginate suspension, the quality-volumetric concentration of described Rhod BH4-sodium alginate suspension is 2～4%；
Step 4, adds the desired amount of calcium chloride solution in the Rhod BH4-sodium alginate suspension of step 3,
Obtain Rhod BH4-sodium alginate porous microsphere；Rhod BH4-sodium alginate suspension by with calcium chloride
Solution reaction forms porous microsphere shape；
Step 5, the Rhod BH4-sodium alginate porous microsphere that step 4 is obtained after over cleaning, dried again
It is placed in calcium chloride solution immersion a period of time.
Wherein, in step 1, described cultivation temperature is 25～35 DEG C, preferably 30 DEG C, and described incubation time is 19～30h,
Preferably 24h.
Wherein, in step 2, in described centrifugal treating, centrifugal force is 10000～15000g, preferably 12000g, centrifugation time
It is 12～20min, preferably 15min.
Wherein, in step 2, the quality-volume of Rhod BH4 suspension described in described Rhod BH4 suspension
Concentration is 180～210mg Rhod BH4/ml water, preferably 200mg Rhod BH4/ml water.
Wherein, in step 3, described Rhod BH4 suspension is 2～5 with the addition volume ratio of sodium alginate suspension:
95～102, preferably 3: 97.
Wherein, in step 3, the quality-volumetric concentration of described Rhod BH4-sodium alginate suspension is 2～4%,
Preferably 4%.
Wherein, in step 4, the quality-volumetric concentration of described calcium chloride solution is 3～5%, preferably 3%.
Wherein, in step 4, the addition of described calcium chloride solution is 10ml.
Wherein, in step 4, the addition speed of described calcium chloride solution is 1.2～1.8ml/min, preferably 1.6ml/min.
Wherein, in step 5, described soak time is 2～5h, preferably 3h.
The Rhod BH4-sodium alginate porous that above-mentioned Rhod BH4-sodium alginate porous microsphere preparation method prepares
Microsphere application in terms of suppression MBR fouling membrane.
In preparation method of the present invention, the formula of LB fluid medium used is: tryptone 10g/L+ yeast extract
5g/L+ sodium chloride 5g/L.
Rhod BH4 used in the present invention extracts from MBR reaction tank, on the one hand illustrates that it can
Surviving in MBR reaction tank, on the other hand, the signaling molecule amount that Rhod BH4 is discharged is the most, it is thus possible to
The enough preferably biomembranous formation of suppression.
The Rhod BH4-sodium alginate porous microsphere that preparation method of the present invention prepares is added in MBR reaction tank
The biomembrane on MBR film surface is carried out lift-off processing, by detection biomembrane charge stripping efficiency, the present invention side of preparation
The Rhod BH4-sodium alginate porous microsphere that method prepares can reach 86.5% to biomembranous charge stripping efficiency.It addition,
In Rhod BH4-sodium alginate porous microsphere, sodium alginate can reach 87.3% to the embedding rate of Rhod BH4,
And form uniformity.Rhod BH4 is wrapped up by sodium alginate, thus plays and delay strain to run off and inactivation
Effect.
Beneficial effect: the preparation method raw material of the present invention is easy to get, low cost, the Rhod BH4-sodium alginate obtained
Porous microsphere has high embedding rate, it is possible to effectively prevent loss and the inactivation of strain, it addition, this Rhod BH4-
Biomembrane is peeled off in terms of physics and biological two by sodium alginate porous microsphere, substantially increases it to biomembranous
Charge stripping efficiency, biomembranous charge stripping efficiency can reach 86.5%, finally this Rhod BH4-sodium alginate porous microsphere
Dependence to response system rate of circulating flow is little.
Detailed description of the invention
According to following embodiment, the present invention be may be better understood.But, as it will be easily appreciated by one skilled in the art that reality
Execute the content described by example and be merely to illustrate the present invention, and should be also without limitation on not described in detail in claims
The present invention.
Embodiment 1
The preparation method of a kind of Rhod BH4-sodium alginate porous microsphere, specifically includes following operating procedure:
Step 1, is inoculated into Rhod BH4 isolated in MBR reaction tank on LB fluid medium, in 25 DEG C
Enrichment culture 19h under constant temperature；
Step 2, is centrifuged 12 minutes under 10000g centrifugal force the Rhod BH4 after enrichment culture, washes with water,
Then it is resuspended in 2ml water being configured to Rhod BH4 suspension；
Step 3, by Rhod BH4 suspension (the mass body volume concentrations of Rhod BH4 suspension of step 2
For 180mg Rhod BH4/ml water) and 95ml sterile sodium alginate suspension be slowly mixed to get Rhod
BH4-sodium alginate suspension, the quality-volumetric concentration of Rhod BH4-sodium alginate suspension is 2% (w/v)；
Step 4, is suspended to the Rhod BH4-sodium alginate of step 3 with the speed of 1.2 ml/min by nozzle
Liquid adds the desired amount of calcium chloride solution (quality-volumetric concentration of calcium chloride solution is 3% (w/v)), obtains Rhodococcus fascians
Belong to BH4-sodium alginate porous microsphere；
Step 5, Rhod BH4-sodium alginate porous microsphere deionized water step 4 obtained cleans three times,
It is dried at room temperature for again, is finally placed in leaching in calcium chloride solution (quality-volumetric concentration of calcium chloride solution is 3% (w/v))
Steep 2 hours.
The Rhod BH4-sodium alginate porous microsphere that embodiment 1 prepares is added in MBR reaction tank MBR film
The biomembrane on surface carries out lift-off processing, by detection biomembrane charge stripping efficiency, the Rhod that embodiment 1 prepares
BH4-sodium alginate porous microsphere is 82.5% to biomembranous charge stripping efficiency.It addition, this Rhod BH4-alginic acid
In sodium porous microsphere, sodium alginate is 82.3% to the embedding rate of Rhod BH4, and form uniformity.
Embodiment 2
The preparation method of a kind of Rhod BH4-sodium alginate porous microsphere, specifically includes following operating procedure:
Step 1, is inoculated into Rhod BH4 isolated in MBR reaction tank on LB fluid medium, in 35 DEG C
Enrichment culture 30h under constant temperature；
Step 2, is centrifuged 20 minutes under 15000g centrifugal force the Rhod BH4 after enrichment culture, washes with water,
Then it is resuspended in 5ml water being configured to Rhod BH4 suspension；
Step 3, by Rhod BH4 suspension (the mass body volume concentrations of Rhod BH4 suspension of step 2
For 210mg Rhod BH4/ml water) and 102ml sterile sodium alginate suspension be slowly mixed to get Rhod
BH4-sodium alginate suspension, the quality-volumetric concentration of Rhod BH4-sodium alginate suspension is 3% (w/v)；
Step 4, is suspended to the Rhod BH4-sodium alginate of step 3 with the speed of 1.7 ml/min by nozzle
Liquid adds the desired amount of calcium chloride solution (quality-volumetric concentration of calcium chloride solution is 5% (w/v)), obtains Rhodococcus fascians
Belong to BH4-sodium alginate porous microsphere；
Step 5, Rhod BH4-sodium alginate porous microsphere deionized water step 4 obtained cleans three times,
It is dried at room temperature for again, is finally placed in leaching in calcium chloride solution (quality-volumetric concentration of calcium chloride solution is 5% (w/v))
Steep 5 hours.
The Rhod BH4-sodium alginate porous microsphere that embodiment 2 prepares is added in MBR reaction tank MBR film
The biomembrane on surface carries out lift-off processing, by detection biomembrane charge stripping efficiency, the Rhod that embodiment 2 prepares
BH4-sodium alginate porous microsphere is 85.0% to biomembranous charge stripping efficiency.It addition, this Rhod BH4-alginic acid
In sodium porous microsphere, sodium alginate is 86.8% to the embedding rate of Rhod BH4, and form uniformity.
Embodiment 3
The preparation method of a kind of Rhod BH4-sodium alginate porous microsphere, specifically includes following operating procedure:
Step 1, is inoculated into Rhod BH4 isolated in MBR reaction tank on LB fluid medium, in 30 DEG C
Enrichment culture 24h under constant temperature；
Step 2, is centrifuged 15 minutes under 12000g centrifugal force the Rhod BH4 after enrichment culture, washes with water,
Then it is resuspended in 3ml water being configured to Rhod BH4 suspension；
Step 3, by Rhod BH4 suspension (the mass body volume concentrations of Rhod BH4 suspension of step 2
For 200mg Rhod BH4/ml water) and 97ml sterile sodium alginate suspension be slowly mixed to get Rhod
BH4-sodium alginate suspension, the quality-volumetric concentration of Rhod BH4-sodium alginate suspension is 4% (w/v)；
Step 4, is suspended to the Rhod BH4-sodium alginate of step 3 with the speed of 1.6 ml/min by nozzle
Liquid adds the desired amount of calcium chloride solution (quality-volumetric concentration of calcium chloride solution is 3% (w/v)), obtains Rhodococcus fascians
Belong to BH4-sodium alginate porous microsphere；
Step 5, Rhod BH4-sodium alginate porous microsphere deionized water step 4 obtained cleans three times,
It is dried at room temperature for again, is finally placed in leaching in calcium chloride solution (quality-volumetric concentration of calcium chloride solution is 3% (w/v))
Steep 3 hours.
The Rhod BH4-sodium alginate porous microsphere that embodiment 3 prepares is added in MBR reaction tank MBR film
The biomembrane on surface carries out lift-off processing, by detection biomembrane charge stripping efficiency, the Rhod that embodiment 3 prepares
BH4-sodium alginate porous microsphere is 86.5% to biomembranous charge stripping efficiency.It addition, this Rhod BH4-alginic acid
In sodium porous microsphere, sodium alginate is 87.3% to the embedding rate of Rhod BH4, and form uniformity.
In the Rhod BH4-sodium alginate porous microsphere that the present invention obtains, sodium alginate is a kind of non-poisonous material to antibacterial
Matter, is used as the material that cell is fixing, and it can reach more than 80% to the embedding rate of strain；Simultaneously because Rhod
(Rhod BH4 persistently produces quorum sensing inhibitor in MBR reaction tank and stops raw in the impact of BH4 colony cancellation
The formation of thing film), decrease the generation of microbial cell biomembrane extracellular polymeric (EPS), thus define a pine
The biomembrane dissipated, makes biomembrane be easier to come off from MBR film surface, it addition, free Rhod BH4-Sargassum
Collision between acid sodium porous microsphere and MBR film produces physical friction effect, further promotes biomembrane from MBR
Film surface comes off, thus substantially increases it to biomembranous charge stripping efficiency.
Claims (10)
1. the preparation method of a Rhod BH4-sodium alginate porous microsphere, it is characterised in that specifically include as follows
Operating procedure:
Step 1, is inoculated into Rhod BH4 isolated in MBR reaction tank on LB fluid medium, Yu Heng
Enrichment culture a period of time under the conditions of temperature；
Step 2, is centrifuged the Rhod BH4 after enrichment culture processing, is configured to Rhod BH4 after being centrifuged
Suspension；
Step 3, is mixed to get Rhod BH4-by the Rhod BH4 suspension of step 2 and sodium alginate suspension
Sodium alginate suspension, the quality-volumetric concentration of described Rhod BH4-sodium alginate suspension is 2～4%；
Step 4, adds the desired amount of calcium chloride solution in the Rhod BH4-sodium alginate suspension of step 3,
Obtain Rhod BH4-sodium alginate porous microsphere；
Step 5, the Rhod BH4-sodium alginate porous microsphere that step 4 is obtained after over cleaning, dried again
It is placed in calcium chloride solution immersion a period of time.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 1, described cultivation temperature is 25～35 DEG C, and described incubation time is 19～30h.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 2, in described centrifugal treating, centrifugal force is 10000～15000g, and centrifugation time is 12～20min.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 2, the quality-volumetric concentration of described Rhod BH4 suspension is 180～210mg Rhod BH4/ml
Water.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 3, described Rhod BH4 suspension is 2～5: 95～102 with the addition volume ratio of sodium alginate suspension.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 4, the quality-volumetric concentration of described calcium chloride solution is 3～5%.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 4, the addition of described calcium chloride solution is 10ml.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 4, the addition speed of described calcium chloride solution is 1.2～1.8ml/min.
The preparation method of Rhod BH4-sodium alginate porous microsphere the most according to claim 1, it is characterised in that:
In step 5, described soak time is 2～5h.
10. the Rhod that Rhod BH4-sodium alginate porous microsphere preparation method described in claim 1 prepares
The application in terms of suppression MBR fouling membrane of the BH4-sodium alginate porous microsphere.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106242076A (en) * | 2016-08-23 | 2016-12-21 | 河海大学 | A kind of microbial film tube bank and its preparation method and application |
| CN106430626A (en) * | 2016-11-11 | 2017-02-22 | 上海海洋大学 | Method for promoting construction of aquaculture bioflocculation process |
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| CN101240270B (en) * | 2007-02-09 | 2010-06-02 | 中国科学院化学研究所 | Alginate capsule immobilized with cell and preparation method thereof |
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2016
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101240270B (en) * | 2007-02-09 | 2010-06-02 | 中国科学院化学研究所 | Alginate capsule immobilized with cell and preparation method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106242076A (en) * | 2016-08-23 | 2016-12-21 | 河海大学 | A kind of microbial film tube bank and its preparation method and application |
| CN106242076B (en) * | 2016-08-23 | 2019-11-12 | 河海大学 | A kind of microbial film tube bank and its preparation method and application |
| CN106430626A (en) * | 2016-11-11 | 2017-02-22 | 上海海洋大学 | Method for promoting construction of aquaculture bioflocculation process |
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