CN1693444A - Process for screening of microorganism flocculator producing bacteria - Google Patents
Process for screening of microorganism flocculator producing bacteria Download PDFInfo
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- CN1693444A CN1693444A CN 200510010779 CN200510010779A CN1693444A CN 1693444 A CN1693444 A CN 1693444A CN 200510010779 CN200510010779 CN 200510010779 CN 200510010779 A CN200510010779 A CN 200510010779A CN 1693444 A CN1693444 A CN 1693444A
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- screening
- microbial flocculant
- congo red
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- flocculant
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Abstract
A method for screening the microbial flocculant generating bacteria features that the Congo red (C32H22N6Na2O6S2) and the Coomassie brilliant blue are simultaneously added to the culture medium for selectively screening the flocculant generating bacteria from different specimens.
Description
Technical field
The present invention relates to a kind of Screening for characteristics microbial flocculant method.
Background technology
Along with development of global economy, the progress of suitability for industrialized production, human standard of living improves greatly, but human environment of depending on for existence but suffers to destroy day by day the disorder of environment rule.The severe situation of water resources shortage and pollution aggravation has promoted the research and the application of water conditioner.On water conditioner flocculation technique development history, developed the flocculation agent (as Tai-Ace S 150, aluminum chloride, iron(ic) chloride etc.) of inorganic salts the sixties, its treatment effect is undesirable; And then the high molecular polymerization inorganic salt type flocculation agent (as polymerize aluminum chloride, bodied ferric sulfate etc.) of development received good effect in water treatment, but can produce certain secondary pollution, especially aluminum ion to environment in its use.Discover that aluminum ions environmental hazard comprises: 1. toxic to hydrobiont, salmon is caused death; 2. influence growth and development of plant, in addition dead; 3. can be by food chain and tap water, finally human health is produced harm, multiple at present senile dementia just be likely because of aluminium content in its brain higher due to.Therefore, various countries have stipulated limit value to aluminium content in the tap water, limited the large-scale application of aluminium salt type flocculation agent, but polyacrylamide is difficult to degraded in environment, easily cause secondary pollution, and monomer whose has intensive neurotoxicity and " three cause " effect (teratogenesis, mutagenesis, carcinogenic), makes its application also restricted greatly, has been used as regulation so it is made.The maximum acceptable concentration of the polyacrylamide SeparunNplo that uses as U.S. approval is 1mg/dm3, and Britain stipulates that the dosage of polyacrylamide on average must not surpass 0.5mg/dm3, and maximum dosage must not surpass 1mg/dm3.Therefore Water Treatment Chemicals new variety, the continuous appearance of new science and technology and the expansion of industrialized scale have greatly been promoted.(Microbial Flocculants MBF) belongs to the biological organic polymer coargulator of the third generation to microbial flocculant.Main component has glycoprotein, polysaccharide, protein, Mierocrystalline cellulose and DNA etc.Microorganism with secretion flocculation agent ability is called bacterium for producing flocculant, finds that so far the microorganism with this flocculence surpasses 17 kinds, comprises mould, bacterium, actinomycetes and yeast etc.Wherein the most representative MBF has following three kinds: the flocculating agent A J7002 that the Aspergillus sojae (Aspergillus sojae) that Nakamura J finds produces; The MBFPF101 that TakagiH produces with variotin (Paecilomyces sp.1-1), it has good flocculating effect to intestinal bacteria, cereuisiae fermentum, active sludge, diatomite etc.; The microbial flocculant NOC-1 that Kurane utilizes rhodococcus erythropolis (Rhodococcus erythropolis) to succeed in developing all has fabulous flocculation and decolorizing effect to slime water, river, water of coal ash, bulking sludge, paper pulp wastewater etc.MBF mainly contains three types: the flocculation agent that directly utilizes microorganism cells; Utilize the flocculation agent of microorganism wall extract; Utilize the flocculation agent of products of cellular metabolism.Characteristics such as MBF has high efficiency, safety non-toxic, non-secondary pollution, the mud generation is little, decolorizing effect is unique, injected volume is few relatively.The prerequisite that obtains highly active microbial flocculant is good production bacterial classification.Normally from different environmental sample separation, purifying microorganism, further the screening back obtains good production bacterial classification.The method of traditional screening bacterium for producing flocculant of microbe mainly is to pass through plate isolation, obtain pure growth, then each pure growth is carried out shake-flask culture, whether the mensuration fermented liquid has flocculation activity is determined bacterium for producing flocculant, exists shortcomings such as the screening target is uncertain, workload is big, the cycle is long, screening efficiency is low.
Summary of the invention
Purpose of the present invention just provides a kind of Screening for characteristics microbial flocculant method, this method can overcome the deficiency of traditional bacterium for producing flocculant of microbe screening method, can be efficiently, intuitively bacterium for producing flocculant and other microorganisms are made a distinction, technology is simple, workload is little, cycle is short, and screening efficiency is higher.
The object of the present invention is achieved like this: this Screening for characteristics microbial flocculant method is to add Congo red (English name: Congo red, molecular formula: C in substratum simultaneously
32H
22N
6Na
2O
6S
2) and Xylene Brilliant Cyanine G, optionally never with screening bacterium for producing flocculant in the sample material.
This Screening for characteristics microbial flocculant method, described Xylene Brilliant Cyanine G are G250 (English name: Coomassie brilliant blue G250, molecular formula: C
47H
48N
3NaO
7S
2).
This Screening for characteristics microbial flocculant method, described Xylene Brilliant Cyanine G are R-250 (English name: Coomassie brilliant blue R-250, molecular formula: C
45H
44N
3NaO
7S
2).
This Screening for characteristics microbial flocculant method, Congo red concentration is 0.0005-0.005% in the substratum., Xylene Brilliant Cyanine G concentration be 0.0005-0.005%.
This Screening for characteristics microbial flocculant method, during medium preparation, Congo red and Xylene Brilliant Cyanine G can be that the corresponding mass fraction of direct weighing adds in the substratum, makes its final concentration reach 0.0005-0.005%.
This Screening for characteristics microbial flocculant method, during medium preparation, Congo red and Xylene Brilliant Cyanine G can be that the form that is mixed with solution adds in the substratum and makes its final concentration reach 0.0005-0.005%.
This Screening for characteristics microbial flocculant method, suitable microorganism comprises bacterium, actinomycetes, yeast and filamentous fungus.
This Screening for characteristics microbial flocculant method is meant and prepares substratum according to a conventional method, adopts dilution spread plate method, or the diluted mixture flat band method, or plate streak carries out separation and the purifying of microorganism, behind single bacterium colony to be formed, and the red bacterium colony of picking.
Screening for characteristics microbial flocculant method of the present invention is owing to be to add Xylene Brilliant Cyanine G and Congo red in substratum, can be efficiently, intuitively bacterium for producing flocculant and other microorganisms are made a distinction, technology is simple, workload is little, cycle is short, and screening efficiency is higher.
The present invention is described in detail below in conjunction with specific embodiment, but the present invention is not limited to by these embodiment again.
Embodiment
Embodiment 1: the screening of flocculation activity bacterium
Get 1000ml bacteria culture medium (g/L: sucrose 10, K
2HPO
41.5, FeCl
30.002, CaCO
31.5, yeast extract paste 0.5, agar 20, water 1000ml, pH7.0～7.5) in a container, add 10mg Xylene Brilliant Cyanine G and Congo red respectively, shake up 121 ℃ of sterilization 20min., make flat board, be coated with different dilution soil supensions, 25-30 ℃ is cultured to dull and stereotyped going up and forms obvious single bacterium colony.Red bacterium colony under the picking blue background inserts 50ml liquid nutrient medium (g/L: sucrose 10, K
2HPO
41.5, FeCl
30.002, CaCO
31.5, yeast extract paste 0.5, pH7.0～7.5) in, cultivate the flocculation activity of measuring fermented liquid after 4 days with the kaolin suspension for 25-30 ℃, 99.6% bacterial strain has flocculation activity as a result.
Embodiment 2: the actinomycetic screening of flocculation activity
Get 1000ml actinomycetes substratum (g/L: Zulkovsky starch 20, KNO
31, NaCl 0.5, K
2HPO
40.5, MgSO
40.5, FeSO
40.01, agar 20, water 1000ml pH7.2-7.4) in a container, adds 10mg Xylene Brilliant Cyanine G and Congo red respectively, shakes up, and 121 ℃ of sterilization 30min. make flat board, are coated with different dilution soil supensions, and 25 ℃ are cultured to dull and stereotyped going up and form obvious single bacterium colony.Red bacterium colony under the picking blue background inserts 50ml liquid nutrient medium (g/L: Zulkovsky starch 20, KNO
31, NaCl 0.5, K
2HPO
40.5, MgSO
40.5, FeSO
40.01, water 1000ml, pH7.2-7.4) in, cultivate the flocculation activity of measuring fermented liquid after 10 days with the kaolin suspension for 25 ℃, 87.9% bacterial strain has flocculation activity as a result.
Embodiment 3: the saccharomycetic screening of flocculation activity
Get 1000ml microzyme culture medium [g/L: yeast extract paste 3, malt extract 3, peptone 5, glucose 20, agar 20, pH5.3 (regulate with hydrochloric acid or phosphoric acid behind the autoclaving, add Broad spectrum antibiotics) to suppress bacterium] in a container, add 10mg Xylene Brilliant Cyanine G and Congo red respectively, shake up, 110 ℃ of sterilization 30min. make flat board, be coated with different dilution soil supensions, 25 ℃ are cultured to dull and stereotyped going up and form obvious single bacterium colony.Red bacterium colony under the picking blue background, insert 50ml liquid nutrient medium [g/L: yeast extract paste 3, malt extract 3, peptone 5, glucose 20, agar 20, pH5.3 (regulating with hydrochloric acid or phosphoric acid behind the autoclaving)] in, cultivate the flocculation activity of measuring fermented liquid after 4 days with the kaolin suspension for 25 ℃, 90.5% bacterial strain has flocculation activity as a result.
Embodiment 4: the screening of flocculation activity filamentous fungus
Get 1000ml fungi culture medium [g/L: glucose 20, peptone 1, yeast extract paste 0.8, KH
2PO
41, MgSO
4.7H
2O 0.5, agar 20, the pH nature, water 1000ml (adding Broad spectrum antibiotics behind the autoclaving) to suppress bacterium] in a container, add 10mg Xylene Brilliant Cyanine G and Congo red respectively, shake up, 110 ℃ of sterilization 30min., make flat board, be coated with different dilution soil supensions, 28 ℃ are cultured to dull and stereotyped going up and form obvious single bacterium colony.Red bacterium colony under the picking blue background inserts 50ml liquid nutrient medium (g/L: glucose 20, peptone 1, yeast extract paste 0.8, KH
2PO
41, MgSO
4.7H
2O 0.5, pH nature, water 1000ml) in, cultivate the flocculation activity of measuring fermented liquid after 4 days with the kaolin suspension for 28 ℃, 82.3% bacterial strain has flocculation activity as a result.
Claims (7)
1, a kind of Screening for characteristics microbial flocculant method is characterized in that adding simultaneously Congo red (English name: Congo red, molecular formula: C in substratum
32H
22N
6Na
2O
6S
2) and Xylene Brilliant Cyanine G, optionally never with screening bacterium for producing flocculant in the sample material.
2, a kind of Screening for characteristics microbial flocculant method as claimed in claim 1 is characterized in that: described Xylene Brilliant Cyanine G is G a 250 (English name: Coomassie brilliant blue G 250, molecular formula: C
47H
48N
3NaO
7S
2).
3, a kind of Screening for characteristics microbial flocculant method as claimed in claim 1 is characterized in that: described Xylene Brilliant Cyanine G is a R-250 (English name: Coomassie brilliant blue R-250, molecular formula: C
45H
44N
3NaO
7S
2).
4, as claim 1,2 or 3 described a kind of Screening for characteristics microbial flocculant methods, it is characterized in that: Congo red concentration is 0.0005-0.005% in the substratum., Xylene Brilliant Cyanine G concentration be 0.0005-0.005%.
5, as claim 1,2 or 3 described a kind of Screening for characteristics microbial flocculant methods, it is characterized in that: during medium preparation, Congo red and Xylene Brilliant Cyanine G can be that the corresponding mass fraction of direct weighing adds in the substratum, makes its final concentration reach 0.0005-0.005%.
6, as claim 1,2 or 3 described a kind of Screening for characteristics microbial flocculant methods, it is characterized in that: during medium preparation, Congo red and Xylene Brilliant Cyanine G can be that the form that is mixed with solution adds in the substratum, makes its final concentration reach 0.0005-0.005%.
7, as claim 1,2 or 3 described a kind of Screening for characteristics microbial flocculant methods, it is characterized in that: the microorganism that is suitable for comprises bacterium, actinomycetes, yeast and filamentous fungus.
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|---|---|---|---|
| CN 200510010779 CN1274805C (en) | 2005-04-29 | 2005-04-29 | Process for screening of microorganism flocculator producing bacteria |
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|---|---|---|---|
| CN 200510010779 CN1274805C (en) | 2005-04-29 | 2005-04-29 | Process for screening of microorganism flocculator producing bacteria |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146797A (en) * | 2013-02-26 | 2013-06-12 | 重庆绿色智能技术研究院 | Compound biological flocculant and preparation method thereof |
| CN103184239A (en) * | 2013-03-11 | 2013-07-03 | 宁波飞日水产实业有限公司 | Method for producing microbial flocculant by using surimi processing waste water |
| CN107043718A (en) * | 2017-03-01 | 2017-08-15 | 沃邦环保有限公司 | It is a kind of for complex micro organism fungicide of river regulation and its preparation method and application |
-
2005
- 2005-04-29 CN CN 200510010779 patent/CN1274805C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146797A (en) * | 2013-02-26 | 2013-06-12 | 重庆绿色智能技术研究院 | Compound biological flocculant and preparation method thereof |
| CN103184239A (en) * | 2013-03-11 | 2013-07-03 | 宁波飞日水产实业有限公司 | Method for producing microbial flocculant by using surimi processing waste water |
| CN107043718A (en) * | 2017-03-01 | 2017-08-15 | 沃邦环保有限公司 | It is a kind of for complex micro organism fungicide of river regulation and its preparation method and application |
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| Publication number | Publication date |
|---|---|
| CN1274805C (en) | 2006-09-13 |
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