CN101407802A - Immobilized preparation of flocculating microbial cell - Google Patents
Immobilized preparation of flocculating microbial cell Download PDFInfo
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- CN101407802A CN101407802A CNA2008100739268A CN200810073926A CN101407802A CN 101407802 A CN101407802 A CN 101407802A CN A2008100739268 A CNA2008100739268 A CN A2008100739268A CN 200810073926 A CN200810073926 A CN 200810073926A CN 101407802 A CN101407802 A CN 101407802A
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- bacterial classification
- immobilized
- calcium chloride
- flocculating
- chloride solution
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Abstract
The invention discloses an immobilized preparation method of a flocculent microbe cell. An effective flocculent microbe is selected as a strain; sodium alginate and calcium chloride are selected as immobilized vectors for obtaining the immobilized flocculent microbe cell grains; and the flocculent activity preservation rate of the immobilized flocculent microbe cell grains achieves 88 percent to 94 percent. The immobilized preparation method is simple and convenient to be operated, has low cost and less harm to the cell; and the flocculent rate of the obtained immobilized grains is high and the effective period is long.
Description
Technical field
The present invention relates to a kind of immobilized cell preparation method, particularly relate to a kind of immobilized preparation of flocculating microbial cell.
Background technology
Microbial flocculant Microbial flocculant is called for short MBF, be a class by the microorganisms justacrine has flocculation activity to the extracellular meta-bolites, have good flocculation sediment performance, safe, nontoxic, be easy to biological degradation.Microbe species that can produce flocculant is many, and growth is fast, is easy to take the biotechnology means to realize industrialization, thereby its research is just being become the important topic of current flocculation agent research.A lot of to biological flocculant research both at home and abroad at present, but mostly research range all is confined to Screening for characteristics microbial flocculant, culture condition to the research of purification, flocculation agent chemical constitution, physico-chemical property and the flocculation mechanism of the influence of its productive rate, microbial flocculant etc., and less for its actual industrialized producing technology research.Though people have found the plurality of advantages of biological flocculant, problems such as its output is little, cost height are used for the practical large-scale of biological flocculant and have been caused huge obstacle.Immobilized biotechnology is a new technology that develops rapidly since the sixties in 20th century.Immobilized cell technology is meant by means chemistry or physics, with the area of space that free cell is positioned to limit, makes it to become and is not suspended in water but still keeps biological activity, and the method that can recycle.Have after cell is immobilized that density height, fast, the anti-murder by poisoning ability of speed of response are strong, product separates easily, can realize operate continuously, can improve advantage such as throughput greatly, therefore obtained developing rapidly and widespread use at immobilized cell technology in recent decades.Therefore, the present invention is directed to immobilized preparation is carried out in a strain from the efficient flocculating microorganism cells of row filter research.
Summary of the invention
The purpose of this invention is to provide and a kind of flocculating microbial cell is carried out immobilized preparation method.
It is bacterial classification that the present invention selects for use a plant height to imitate flocculating microbial, cultivates 24 hours at 30 ℃ of 150r/min, and the culture of strains base consists of glucose 20g, KH
2PO
42g, K
2HPO
45g, (NH
4)
2SO
40.2g, NaCl 0.1g, urea 0.5g, yeast extract paste 0.5g, MgSO
47H
2O 0.2g, distilled water 1000ml, pH 7.2-7.6; With liquid medium centrifugal 30min under 3000r/m, use the stroke-physiological saline solution dissolution precipitation, constant volume is to former scale, and is centrifugal under the same conditions, gets wet thallus.The sodium alginate of getting the 100-150ml mass percent concentration and be 2%-6% mixes with the 0.4-0.8g wet thallus and stirs.The preparation mass percent concentration is the calcium chloride solution 100ml-200ml of 7%-13%, place 0 ℃ mixture of ice and water, with syringe sodium alginate-bacterial classification mixed solution drop is splashed in the calcium chloride solution, form spherical particle, keep 1h-2h, make the abundant immobilization of bacterial classification at 10 ℃-25 ℃, remove supernatant liquor, spherical gel bacterial classification after the water flushing immobilization places calcium chloride solution balance 24h, i.e. the flocculating microbial cell particle of being fixed then again.Bacterial classification after the immobilization is carried out flocculating rate measure, active storage rate reaches 88%-94%.
The measuring method of flocculating rate is, in the 100ml graduated cylinder, add people 80ml distilled water, 0.4g kaolin, the CaCl solution of 5ml 1%, 2ml supernatant liquor testing sample, adding distil water is to 100ml adjusting pH to 7.0 again, pour into then in the 150ml beaker, be placed on and stir 1min on the magnetic stirring apparatus fast, stir 5min at a slow speed, leave standstill 10min, draw the supernatant liquor of certain depth with suction pipe and measure absorbancy in spectrophotometer 550nm place, simultaneously compare experiment, and calculate its flocculating rate with following formula with the supernatant liquor that does not add flocculation agent:
Flocculating rate=(A-B)/A * 100%
In the formula: A-contrast supernatant liquor is in the absorbancy at 550nm place
B-sample supernatant liquor is in the absorbancy at 550nm place
The present invention is easy and simple to handle, and cost is cheap, and the infringement of pair cell is little, the immobilization particle flocculating rate height that obtains, and validity period is long.
Embodiment
Embodiment 1:
Selecting for use a plant height to imitate flocculating microbial is bacterial classification, cultivates 24 hours at 30 ℃ of 150r/min, and the culture of strains base consists of glucose 20g, KH
2PO
42g, K
2HPO
45g, (NH
4)
2SO
40.2g, NaCl 0.1g, urea 0.5g, yeast extract paste 0.5g, MgSO
47H
2O 0.2g, distilled water 1000ml, pH 7.2-7.6; With liquid medium centrifugal 30min under 3000r/m, use the stroke-physiological saline solution dissolution precipitation, constant volume is to former scale, and is centrifugal under the same conditions, gets wet thallus.Getting mass percent concentration is 3% sodium alginate 40ml, mixes and adds sterilized water with the 0.13g bacterial classification and stir to 50ml.The preparation mass percent concentration is 10% calcium chloride solution 100ml, place 0 ℃ mixture of ice and water, with syringe sodium alginate-bacterial classification mixed solution drop is splashed in the calcium chloride solution, form spherical particle, keep 1h, make the abundant immobilization of bacterial classification at 15 ℃, remove supernatant liquor, spherical gel bacterial classification after the water flushing immobilization places calcium chloride solution balance 24h, i.e. the flocculating microbial cell particle of being fixed then again.Bacterial classification after the immobilization is carried out flocculating rate measure, active storage rate reaches 92.4%.
Embodiment 2:
Selecting for use a plant height to imitate flocculating microbial is bacterial classification, cultivates 24 hours at 30 ℃ of 150r/min, with liquid medium centrifugal 30min under 3000r/m, gets thalline, and the culture of strains base consists of glucose 20g, KH
2PO
42g, K
2HPO
45g, (NH
4)
2SO
40.2g, NaCl 0.1g, urea 0.5g, yeast extract paste 0.5g, MgSO
47H
2O0.2g, distilled water 1000ml, pH 7.2-7.6; Get 100g thalline stroke-physiological saline solution dissolution precipitation.Getting the 450g sodium alginate mixes and adds sterilized water with sterilized water dissolving back and stir to 15L with bacterial classification.The preparation mass percent concentration is 10% calcium chloride solution 10L, place 0 ℃ mixture of ice and water, with syringe sodium alginate-bacterial classification mixed solution drop is splashed in the calcium chloride solution, form spherical particle, keep 1h, make the abundant immobilization of bacterial classification at 25 ℃, remove supernatant liquor, spherical gel bacterial classification after the water flushing immobilization places calcium chloride solution balance 24h, i.e. the flocculating microbial cell particle of being fixed then again.Bacterial classification after the immobilization is carried out flocculating rate measure, active storage rate reaches 90%.
Claims (1)
1. the immobilized preparation of a flocculating microbial cell is characterized in that step is: selecting for use a plant height to imitate flocculating microbial is bacterial classification, cultivates 24 hours at 30 ℃ of 150r/min, and the culture of strains base consists of glucose 20g, KH
2PO
42g, K
2HPO
45g, (NH
4)
2SO
40.2g, NaCl 0.1g, urea 0.5g, yeast extract paste 0.5g, MgSO
47H
2O 0.2g, distilled water 1000ml, pH 7.2-7.6; With liquid medium centrifugal 30min under 3000r/m, use the stroke-physiological saline solution dissolution precipitation, constant volume is to former scale, and is centrifugal under the same conditions, gets wet thallus; The sodium alginate of getting the 100-150ml mass percent concentration and be 2%-6% mixes with the 0.4-0.8g wet thallus and stirs; The preparation mass percent concentration is the calcium chloride solution 100ml-200ml of 7%-13%, place 0 ℃ mixture of ice and water, with syringe sodium alginate-bacterial classification mixed solution drop is splashed in the calcium chloride solution, form spherical particle, keep 1h-2h, make the abundant immobilization of bacterial classification at 10 ℃-25 ℃, remove supernatant liquor, spherical gel bacterial classification after the water flushing immobilization places calcium chloride solution balance 24h, i.e. the flocculating microbial cell particle of being fixed then again.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100739268A CN101407802A (en) | 2008-11-18 | 2008-11-18 | Immobilized preparation of flocculating microbial cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100739268A CN101407802A (en) | 2008-11-18 | 2008-11-18 | Immobilized preparation of flocculating microbial cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101407802A true CN101407802A (en) | 2009-04-15 |
Family
ID=40571014
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|---|---|---|---|
| CNA2008100739268A Pending CN101407802A (en) | 2008-11-18 | 2008-11-18 | Immobilized preparation of flocculating microbial cell |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104277997A (en) * | 2014-08-15 | 2015-01-14 | 安徽省农业科学院烟草研究所 | Fast separation and screening and storage method for tobacco stem hollow germs |
| CN109321558A (en) * | 2018-10-31 | 2019-02-12 | 武汉工程大学 | The method for preparing phosphate solubilizing microorganism sustained release sodium alginate micro ball using emulsion process |
| CN110734905A (en) * | 2019-11-20 | 2020-01-31 | 广东希普生物科技股份有限公司 | Immobilized microorganism particles, preparation method thereof and water quality purification method |
-
2008
- 2008-11-18 CN CNA2008100739268A patent/CN101407802A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104277997A (en) * | 2014-08-15 | 2015-01-14 | 安徽省农业科学院烟草研究所 | Fast separation and screening and storage method for tobacco stem hollow germs |
| CN104277997B (en) * | 2014-08-15 | 2018-02-23 | 安徽省农业科学院烟草研究所 | One quick separating for growing tobacco hollw stalk bacterium screens and store method |
| CN109321558A (en) * | 2018-10-31 | 2019-02-12 | 武汉工程大学 | The method for preparing phosphate solubilizing microorganism sustained release sodium alginate micro ball using emulsion process |
| CN109321558B (en) * | 2018-10-31 | 2022-12-06 | 武汉工程大学 | Method for preparing phosphate-solubilizing microorganism slow-release sodium alginate microspheres by using emulsification method |
| CN110734905A (en) * | 2019-11-20 | 2020-01-31 | 广东希普生物科技股份有限公司 | Immobilized microorganism particles, preparation method thereof and water quality purification method |
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| PB01 | Publication | ||
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Open date: 20090415 |