CN100334012C - Method for preparing biological flocculant from arthrobacter - Google Patents
Method for preparing biological flocculant from arthrobacter Download PDFInfo
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- CN100334012C CN100334012C CNB200310105515XA CN200310105515A CN100334012C CN 100334012 C CN100334012 C CN 100334012C CN B200310105515X A CNB200310105515X A CN B200310105515XA CN 200310105515 A CN200310105515 A CN 200310105515A CN 100334012 C CN100334012 C CN 100334012C
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- arthrobacter
- biological flocculant
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- hpo
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Abstract
The present invention provides a method for preparing biological flocculating agents by using arthrobacteria, which comprises fermentation and cultivation of strains and preparation of biological flocculating agents. Arthrobacteria are adopted as strains of biological flocculating agents to obtain biological flocculating agents for water treatment through fermentation, cultivation and extraction. Compared with the prior art, the method has the characteristics of short production period, high flocculation efficiency, no secondary pollution of use, suitability for mass production and industrial application, etc., thereby having good popularization value.
Description
One, technical field
The present invention relates to the preparation method of a kind of water treatment, specifically a kind of method of utilizing Arthrobacter to prepare biological flocculant with flocculation agent.
Two, technical background
It is the inorganic polymer class of representative that the present flocculation agent that uses in feedwater and sewage disposal has with aluminium, with the polyacrylamide is the synthetic organic polymer class of representative, studies show that, among the aluminum ions crowd of tap water Excessive Intake, the ratio of senile dementia is higher, and the polyacrylamide amine monomers has intensive neurotoxin and carcinogenesis.Though the chitosan that extracts from chitin can be used as flocculation agent, complex manufacturing, resource are very limited.Microbial flocculant is to utilize biotechnology to pass through novel, the efficient water conditioner that strain fermentation is produced.Except that the effect that possesses the traditional chemical synthetic flocculant, that microbial flocculant also has is nontoxic, safe in utilization, have biodegradability, raw materials for production such as are easy to get, can be mass-produced at advantage.Just development in science and technology trend is seen at present, and microbial flocculant has the trend of the synthetic like product of instead of chemical.The bacterium for producing flocculant of microbe of having reported relates to bacterium, actinomycetes, mould song and yeast, and fermentative production all exists raw materials cost height, fermentation period is long, flocculation efficiency is low deficiency.
Three, summary of the invention
The objective of the invention is to overcome above flocculation agent preparation method's deficiency, provide a kind of with short production cycle, flocculation efficiency is high, can adapt to scale operation and industrial application prepare the method for biological flocculant by Arthrobacter.
Four, embodiment
The objective of the invention is to realize in the following manner, selecting Arthrobacter for use is the biological flocculant bacterial classification, makes the water-treatment biological flocculation agent through fermentation culture and extraction, and following steps are adopted in the preparation of this biological flocculant:
A) preparation of bacterial classification
Arthrobacter Arthrobacter Sp.LF-Tou2 is obtained by soil sampling, separation.
Optionally substratum weight is formed g/L:
Hexanaphthene 5-15 (NH
4)
2SO
42-7 K
2HPO
41-5 KH
2PO
41-4
MgSO
40.1-0.4 NaCl 0.05-0.3 agar powder 10-20 pH 7.0-7.4.
The weight of final election substratum is formed g/L:
Glucose 7-12 (NH
4)
2SO
42-5 K
2HPO
41-5
KH
2PO
4 1-4 MgSO
4 0.1-0.5 NaCl 0.1-0.3
Add agar powder 10-20 pH 7.0-7.4 during as solid culture
Inoculation back culture condition is 170 rev/mins, and 30 ℃ of shaking tables were cultivated 1-2 days, measured the flocculation activity of fermented liquid.
First choosing method: it is dull and stereotyped to select substratum to make with selectivity, with soil extract coating inoculation.30 ℃ of incubators were cultivated 1-7 days, and picking growth bacterium colony fast, that transparent circle is big changes in the solid inclined-plane, 30 ℃ of incubators were cultivated 4 ℃ of preservations 1-3 days.
The physiological and biochemical property of Arthrobacter is:
Can clear up polychrom, reduction nitrate has glucosidase activity, can utilize nutritive ingredient growth and breedings such as inorganic nitrogen, citric acid, glucose, wood sugar, sucrose, cellobiose, fructose, semi-lactosi.Can not hydrolyzed starch, Mierocrystalline cellulose.
B) strain fermentation is cultivated
The Arthrobacter bacterial classification is inoculated in the ratio of the 1-2% of nutrient solution gross weight stirs fermentation in the fermentor tank of being furnished with liquid fermentation medium, leavening temperature is 24-32 ℃, blasting air capacity is 0.25-1.5L/L. minute, pH=7.0-7.4, and stir culture 1-2 days are standby;
Liquid fermentation medium is made up of following weight proportion raw material:
Glucose 8-15 part, (NH
4)
2SO
41-5 part, K
2HPO
40.1-1 part, KH
2PO
40.05-1 part, MgSO
40.1-1 part, NaCl 0.2-1.5 part, CaCl
20.2-1.5 part, 1000 parts in water.
C) preparation of biological flocculant:
(1) fermented liquid is removed precipitation by whizzer centrifugation 10-40 minute, and it is standby to get supernatant liquor;
(2) dehydrated alcohol of adding 0.05-0.15% calcium ion and 2.5 times of volumes in supernatant liquor, room temperature left standstill 30-60 minute, used the whizzer centrifugation again 10-20 minute, and it is standby to get flocculation agent crude product throw out;
(3) throw out 0.5-3.0%, the sodium chloride solution dissolving of pH=5-6, with the cetyl trimethylammonium bromide absorption of 0.3-1 times of throw out volume, it is standby to get throw out through centrifugation again;
(4) will precipitate with the dissolving of the sodium-chlor of 5-15% and stir 6-18 hour, add 2.5-3.5 times of volume of ethanol, stir, leave standstill 20-30 minute, centrifugal must throw out, throw out washed with vacuum-drying with dehydrated alcohol alcohol gets the biological flocculant product.
The preferred weight proportioning that the liquid fermentation medium raw material is formed is:
Glucose 10-12 part, (NH
4)
2SO
42-3 part, K
2HPO
40.3-0.8 part, KH
2PO
40.1-0.6 part, MgSO
40.3-0.6 part, NaCl 0.5-1 part, CaCl
20.5-1 part, 1000 parts in water.
The optimum weight proportioning that the liquid fermentation medium raw material is formed is:
11 parts of glucose, (NH
4)
2SO
42.5 part, K
2HPO
40.6 part, KH
2PO
40.4 part, MgSO
40.5 part, 0.7 part of NaCl, CaCl
20.8 part, 1000 parts in water.
Rotating speed is controlled at 5000-10000 rev/min when using whizzer to separate.
The method for preparing biological flocculant by Arthrobacter of the present invention compare with the preparation method of various flocculation agents in the prior art flocculation agent flocculation efficiency height that has that raw materials cost is low, technology is simple, fermentation period is short, generate, nontoxic, use non-secondary pollution, adapt to characteristics such as scale operation and industrial application, have good value for applications.
Four, embodiment:
The preparation of bacterial classification
Obtain Arthrobacter Arthrobacter Sp.LF-Tou2 by soil sampling, separation
Bacterial classification choosing choosing:
Selectivity spawn culture based formulas g/L:
Hexanaphthene 10 (NH
4)
2SO
45 K
2HPO
43 KH
2PO
42
MgSO
40.2 NaCl 0.1 agar powder 15 pH 7.0-7.4.
Final election spawn culture based formulas g/L:
Glucose 10 (NH
4)
2SO
43 K
2HPO
43
KH
2PO
4 2 MgSO
4 0.2 NaCl 0.1
Add agar powder 15 pH 7.0-7.4 when cultivating as solid spawn
Inoculation back culture of strains condition is 170 rev/mins, and 30 ℃ of shaking tables were cultivated 1-2 days, measured the fermented liquid flocculation activity.
The primary election of bacterial classification:
It is dull and stereotyped to select substratum to make with selectivity, with soil extract coating inoculation.30 ℃ of incubators were cultivated 1-7 days, and picking growth bacterium colony fast, that transparent circle is big changes in the solid inclined-plane, 30 ℃ of incubators were cultivated 4 ℃ of preservations 1-3 days.
The final election of bacterial classification:
The triangular flask that the final election substratum is arranged in the picking lawn is inoculated in from the inclined-plane, 30 ℃, 170 rev/mins of shaking tables were cultivated I-2 days, detected the activity of fermented liquid, selected the high bacterial strain that produces of biological flocculant.
The detection of flocculation activity:
In the 100ml measuring graduates, add 0.4% kaolin suspension 97ml, 5%CaCl
22ml, transferring pH with 5%NaOH is 7.5, again with fermented liquid 0.3ml, supplies with distilled water and to be 100ml, back and forth 30 seconds of mixing, staticly settles 5 minutes, getting scale is the supernatant mensuration optical density(OD) at 50ml place.
The preparation of biological flocculant
Get glucose 14kg, (NH
4)
2SO
42kg, K
2HPO
40.4kg, KH
2PO
40.3kg, MgSO
40.4kg, NaCl 0.3kg, CaCl
20.5kg place water 1000kg water to be made into liquid fermentation medium.Ratio according to nutrient solution gross weight 2% down stirs fermentation at 28 ℃ with the Arthrobacter bacterial classification inoculation in the fermentor tank that fills the aforesaid liquid fermention medium, cultivated 1-2 days.Control blasts air capacity at 0.75L/L.min, pH=7.0 in the fermenting process.
The fermented liquid that obtains previously by whizzer centrifugation 30 minutes, is got supernatant liquor and add the dehydrated alcohol of 0.10% calcium ion and 2.5 times of volumes in clear liquid, and room temperature left standstill 30 minutes, used the whizzer centrifugation again 15 minutes, flocculation agent crude product throw out.Throw out is with 1.5%, after the sodium chloride solution dissolving of pH=5.5, cetyl trimethylammonium bromide absorption with 0.6 times of throw out volume, get throw out through centrifugation again, again this throw out was stirred 12 hours with 10% sodium-chlor dissolving, add 3 times of volume of ethanol, stir, left standstill 20 minutes, centrifugal throw out is washed throw out with vacuum-drying with dehydrated alcohol alcohol and to be got the biological flocculant product.
Centrifuge speed all is controlled at 5000-7000 rev/min in the centrifugal separation processes.
The biological flocculant of being produced by method of the present invention is applicable to the various sewage disposals that contain particulate contamination or pollutent, for example sanitary sewage, the sewage of foodstuffs industry discharging, the purifying treatment of surface water especially have treatment effect preferably to the sewage of pH value in the 6-9 scope.Using method is identical with the using method of other water treatment flocculant.
Claims (3)
1, the method for preparing biological flocculant by Arthrobacter, it is characterized in that selecting for use Arthrobacter is the biological flocculant bacterial classification, makes the water-treatment biological flocculation agent through fermentation culture and extraction, following steps are adopted in the preparation of this biological flocculant:
A) preparation of bacterial classification
Obtain Arthrobacter (Arthrobacter Sp.LF-Tou2) by soil sampling, separation;
Optionally substratum weight is formed g/L:
Hexanaphthene 5-15 (NH
4)
2SO
42-7 K
2HPO
41-5 KH
2PO
41-4
MgSO
40.1-0.4 NaCl 0.05-0.3 agar powder 10-20 pH 7.0-7.4
The weight of final election substratum is formed g/L:
Glucose 7-12 (NH
4)
2SO
42-5 K
2HPO
41-5
KH
2PO
4?1-4 MgSO
4 0.1-0.5 NaCl 0.1-0.3
Add agar powder 10-20 pH 7.0-7.4 during as solid culture
Inoculation back culture condition is 170 rev/mins, and 30 ℃ of shaking tables were cultivated 1-2 days, measured the flocculation activity of fermented liquid;
First choosing method: it is dull and stereotyped to select substratum to make with selectivity, and with soil extract coating inoculation, 30 ℃ of incubators were cultivated 1-7 days, and picking growth bacterium colony fast, that transparent circle is big changes in the solid inclined-plane, 30 ℃ of incubators cultivations 1-3 days, 4 ℃ of preservations;
B) strain fermentation is cultivated
Be inoculated in stirring fermentation in the fermentor tank of being furnished with liquid fermentation medium with just selecting the ratio of Arthrobacter bacterial classification in the 1-2% of nutrient solution gross weight, leavening temperature is 24-32 ℃, blasting air capacity is 0.25-1.5L/L. minute, pH=7.0-7.4, and stir culture 1-2 days are standby;
C) preparation of biological flocculant:
Liquid fermentation medium is made up of following weight proportion raw material:
Glucose 8-15 part, (NH
4)
2SO
41-5 part, K
2HPO
40.1-1 part, KH
2PO
40.05-1 part, MgSO
40.1-1 part, NaCl 0.2-1.5 part, CaCl
20.2-1.5 part, 1000 parts in water;
(1) fermented liquid is removed precipitation by whizzer centrifugation 10-40 minute, and it is standby to get supernatant liquor;
(2) dehydrated alcohol of adding 0.05-0.15% calcium ion and 2.5 times of volumes in supernatant liquor, room temperature left standstill 30-60 minute, used the whizzer centrifugation again 10-20 minute, and it is standby to get flocculation agent crude product throw out;
(3) throw out 0.5-3.0%, the sodium chloride solution dissolving of pH=5-6, with the cetyl trimethylammonium bromide absorption of 0.3-1 times of throw out volume, it is standby to get throw out through centrifugation again;
(4) will precipitate with the dissolving of the sodium-chlor of 5-15% and stir 6-18 hour, add 2.5-3.5 times of volume of ethanol, stir, leave standstill 20-30 minute, centrifugal must throw out, throw out washed with vacuum-drying with dehydrated alcohol alcohol gets the biological flocculant product.
2, according to claim 1ly prepare the method for biological flocculant, it is characterized in that liquid fermentation medium is made up of following weight proportion raw material by Arthrobacter:
Glucose 10-12 part, (NH
4)
2SO
42-3 part, K
2HPO
40.3-0.8 part, KH
2PO
40.1-0.6 part, MgSO
40.3-0.6 part, CaCl
20.5-1 part, 1000 parts in water.
3, according to claim 1ly prepare the method for biological flocculant, it is characterized in that the rotating speed of whizzer is controlled at 5000-10000 rev/min by Arthrobacter.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200310105515XA CN100334012C (en) | 2003-11-13 | 2003-11-13 | Method for preparing biological flocculant from arthrobacter |
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|---|---|---|---|
| CNB200310105515XA CN100334012C (en) | 2003-11-13 | 2003-11-13 | Method for preparing biological flocculant from arthrobacter |
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|---|---|
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| CN100334012C true CN100334012C (en) | 2007-08-29 |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100532288C (en) * | 2007-06-18 | 2009-08-26 | 同济大学 | Method for preparing biological flocculant using Proteus mirabilis |
| CN100549159C (en) * | 2007-08-01 | 2009-10-14 | 陕西省科学院酶工程研究所 | A kind of preparation method of biological flocculant |
| CN103146797B (en) * | 2013-02-26 | 2015-04-15 | 中国科学院重庆绿色智能技术研究院 | Compound biological flocculant and preparation method thereof |
| CN104528952B (en) * | 2014-07-21 | 2016-07-20 | 济南大学 | A kind of effective microorganisms processing not calcium ions strong basicity waste water and application thereof |
| CN105176877B (en) | 2015-10-12 | 2017-05-17 | 青岛耀东生物工程有限公司 | Klebsiella and method for preparing microbial flocculant by using same |
| CN105712494B (en) * | 2016-04-14 | 2018-03-13 | 太原理工大学 | A kind of method of microbial flocculant and surfactant coagulating treatment mining industry waste water |
| CN106520595B (en) * | 2016-10-14 | 2019-10-08 | 安徽省农业科学院园艺研究所 | A kind of arthrobacterium and its application in terms of biological control bacterial wilt of tomato |
| CN106365279A (en) * | 2016-10-16 | 2017-02-01 | 谭淞文 | Microbiological product capable of flocculation and coagulation enhancement and preparation method, use method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4146706A (en) * | 1976-01-19 | 1979-03-27 | Idemitsu Kosan Company Limited | Polysaccharide and process for the production thereof |
| CN1322681A (en) * | 2001-01-11 | 2001-11-21 | 无锡轻工大学 | Biological flocculant prepared with Nocardia bacteria |
-
2003
- 2003-11-13 CN CNB200310105515XA patent/CN100334012C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4146706A (en) * | 1976-01-19 | 1979-03-27 | Idemitsu Kosan Company Limited | Polysaccharide and process for the production thereof |
| CN1322681A (en) * | 2001-01-11 | 2001-11-21 | 无锡轻工大学 | Biological flocculant prepared with Nocardia bacteria |
Non-Patent Citations (1)
| Title |
|---|
| 耐盐降解苯乙酸类菌株A1（Arthrobacter sp.A1）的分离和特性研究 何健 崔中利 刘智 李顺鹏，环境科学学报，第22卷第3期 2002 * |
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