CN105039415A - Method for synthesizing flocculant by utilizing Acinetobacter baumanii - Google Patents
Method for synthesizing flocculant by utilizing Acinetobacter baumanii Download PDFInfo
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- CN105039415A CN105039415A CN201510355798.6A CN201510355798A CN105039415A CN 105039415 A CN105039415 A CN 105039415A CN 201510355798 A CN201510355798 A CN 201510355798A CN 105039415 A CN105039415 A CN 105039415A
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- Prior art keywords
- flocculant
- congo red
- centrifugal
- acinetobacter bauamnnii
- bacterium liquid
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Links
- 238000000034 method Methods 0.000 title claims abstract description 22
- 241000588626 Acinetobacter baumannii Species 0.000 title abstract description 5
- 230000002194 synthesizing effect Effects 0.000 title abstract 2
- 238000005189 flocculation Methods 0.000 claims abstract description 25
- 230000016615 flocculation Effects 0.000 claims abstract description 24
- IQFVPQOLBLOTPF-HKXUKFGYSA-L congo red Chemical compound [Na+].[Na+].C1=CC=CC2=C(N)C(/N=N/C3=CC=C(C=C3)C3=CC=C(C=C3)/N=N/C3=C(C4=CC=CC=C4C(=C3)S([O-])(=O)=O)N)=CC(S([O-])(=O)=O)=C21 IQFVPQOLBLOTPF-HKXUKFGYSA-L 0.000 claims abstract description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000002351 wastewater Substances 0.000 claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 10
- 241000894006 Bacteria Species 0.000 claims abstract description 9
- 239000006228 supernatant Substances 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 6
- 239000000047 product Substances 0.000 claims abstract description 4
- 241000589291 Acinetobacter Species 0.000 claims description 19
- 239000003795 chemical substances by application Substances 0.000 claims description 19
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 230000004913 activation Effects 0.000 claims description 5
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 4
- 238000011081 inoculation Methods 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- 229920001817 Agar Polymers 0.000 claims description 2
- 239000001888 Peptone Substances 0.000 claims description 2
- 108010080698 Peptones Proteins 0.000 claims description 2
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- 239000008272 agar Substances 0.000 claims description 2
- 230000000721 bacterilogical effect Effects 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 claims description 2
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 2
- 235000019319 peptone Nutrition 0.000 claims description 2
- -1 polyoxyethylene Polymers 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 6
- 150000004676 glycans Chemical class 0.000 abstract description 6
- 229920001282 polysaccharide Polymers 0.000 abstract description 6
- 239000005017 polysaccharide Substances 0.000 abstract description 6
- 238000001556 precipitation Methods 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 2
- 238000005119 centrifugation Methods 0.000 abstract 3
- 238000004088 simulation Methods 0.000 abstract 2
- 239000002202 Polyethylene glycol Substances 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 abstract 1
- 238000004108 freeze drying Methods 0.000 abstract 1
- 239000002075 main ingredient Substances 0.000 abstract 1
- 229920001223 polyethylene glycol Polymers 0.000 abstract 1
- 230000003311 flocculating effect Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000002207 metabolite Substances 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000008394 flocculating agent Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229920002907 Guar gum Polymers 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229940045110 chitosan Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 229940014259 gelatin Drugs 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 238000004065 wastewater treatment Methods 0.000 description 1
Abstract
The invention discloses a method for synthesizing a flocculant by utilizing Acinetobacter baumanii. The method comprises the following steps: S1, strain activization: inoculating Acinetobacter baumanii (/i) YNWH 226 in a solid medium for activization; S2, cultivation: inoculating a single colony obtained through activization into Congo red simulation waste water for cultivation, so as to obtain a bacterium liquid; S3, centrifugation: conducting centrifugation on the bacterium liquid obtained in the S2, and taking the supernatant after centrifugation to be mixed with ethyl alcohol or poly ethylene glycol; S4, conducting freeze drying on the precipitation product obtained in the S3, so as to obtain the flocculant. The flocculant, of which the main ingredient is polysaccharides, is synthesized through cultivation of Acinetobacter baumanii (/i) YNWH 226 in Congo red simulation waste water. The method has the advantages that the flocculant preparation cost is relatively low; the efficient flocculation pH range is relatively wide; the flocculation effect is relatively good; the industrial application potentiality is high.
Description
Technical field
The invention belongs to biotechnology, field of environment engineering technology, particularly, relate to a kind of method utilizing Acinetobacter bauamnnii synthetic flocculant.
Background technology
Flocculation agent is divided into organic floculant, inorganic flocculating agent, biological flocculant.Traditional organic floculant and inorganic flocculating agent are widely used in the flocculation process of wastewater treatment, although it has higher cost performance, itself are not easy maybe to be harmful to biology in nature degraded.In recent years, the high efficiency because of biological flocculant and the innocuousness to environment, receive the concern of numerous investigator.Common biological flocculant has chitosan, guar gum, sodium alginate, gelatin and microbial flocculant.But the shortcomings such as traditional biological flocculant also also exists, and production cost is high, the production time is long.
Microbial flocculant is the meta-bolites with flocculation activity that a class is produced by microorganism, and main component is polysaccharide, protein, lipid etc.Extracellular polymeric is main microbial flocculant is the special meta-bolites produced by microorganism, the water treatment agent that the class obtained by fermentable, separation, extraction is new and effective.The preparation method of existing microbial flocculant often exists that production cost is high, the production cycle is long, and the flocculation agent prepared effectively flocculates pH narrow range, and the problems such as flocculating effect is poor, are difficult to apply.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, provide a kind of method utilizing Acinetobacter bauamnnii synthetic flocculant.The present invention chooses Acinetobacter bauamnnii, and utilize cheap Congo red waste water replace expensive culture medium culturing and collect its meta-bolites, produce the flocculation agent cost obtained lower, effectively flocculation pH scope is comparatively wide, and flocculating effect is better.
Above-mentioned purpose of the present invention is achieved by the following technical programs.
Utilize a method for Acinetobacter bauamnnii synthetic flocculant, comprise the steps:
S1. actication of culture: by Acinetobacter bauamnnii
acinetobacterbaumanniiyNWH226 is inoculated in solid medium and activates;
S2. cultivate: single colony inoculation activation obtained, in Congo red simulated wastewater, is cultivated and obtained bacterium liquid;
S3. the bacterium liquid obtained by S2 is centrifugal, get centrifugal after supernatant liquor mix with ethanol or polyoxyethylene glycol;
S4. get the precipitated product that S3 obtains and carry out lyophilize, obtain flocculation agent;
Wherein, Acinetobacter bauamnnii described in S1
acinetobacterbaumanniithe preserving number of YNWH226 is CGMCC7922;
Described in S2, Congo red simulated wastewater comprises: K
2hPO
41.6g/L, KH
2pO
40.2g/L, (NH
4)
2sO
41.0g/L, FeSO
47H
2o0.01g/L, NaCl0.1g/L, CaCl
26H
2o0.02g/L, MgSO
40.1g/L, glucose 1g/L, pH7.0, Congo red to 100mg/L after adding filtration sterilization after sterilizing in above-mentioned substratum.
The present invention select a strain Acinetobacter bauamnnii (
acinetobacterbaumannii), by name
acinetobacterbaumanniiyNWH226, be preserved in Beijing China General Microbiological culture presevation administrative center on July 15th, 2013, preserving number is CGMCC7922.It is sole carbon source and energy growth and breeding that this bacterial strain can utilize azoic dyestuff Congo red, can decolour completely under the condition of purebred cultivation by Congo red in minimal medium.The present invention selects cheap Congo red simulated wastewater to cultivate Acinetobacter bauamnnii
acinetobacterbaumanniiyNWH226, be not because it can make Acinetobacter bauamnnii YNWH226 growth and breeding thus obtain more bacterium liquid cell better relative to other substratum (as LB liquid nutrient medium), but because contriver finds that the culture condition of Congo red simulated wastewater can stimulate the Acinetobacter bauamnnii YNWH226 of Board Lot to secrete more extracellular polymeric polysaccharide, and using when extracting the extracellular polymeric polysaccharide that obtains as flocculation agent, find that its flocculation agent effect is better than other substratum.The Acinetobacter bauamnnii that the present invention chooses, not yet has bibliographical information using its meta-bolites as the research of flocculation agent.
Preferably, described in S1, solid medium comprises: bacteriological peptone 10g/L, yeast powder 5g/L, NaCl10g/L, agar powder 15g/L, pH7.0.
Preferably, the time activated described in S1 is 24h.
Preferably, the time of cultivating described in S2 is 48h.
Preferably, centrifugal described in S3 is the centrifugal 30min of 8000g.
Preferably, the volume ratio hybrid reaction 2h of 1:2 pressed by supernatant liquor described in S3 and ethanol.When the volume ratio of supernatant liquor and ethanol is lower than 1:2, polysaccharide flocculant cannot effectively precipitate; When volume ratio is higher than 1:2, cause the waste of ethanol.
Preferably, cryodesiccated condition described in S4 is-20 DEG C or following lyophilize 12h.
Preferably, described in S4, cryodesiccated temperature is-32 DEG C.
Compared with prior art, beneficial effect of the present invention is: the present invention selects Acinetobacter bauamnnii
acinetobacterbaumanniiyNWH226, utilizes Congo red simulated wastewater, and synthesis main component is the flocculation agent of polysaccharide.Flocculation agent cost lower, the effective flocculation pH scope that this method is produced comparatively wide (pH is effective in 5 ~ 8 scopes), flocculating effect are better, and industrial application potentiality are large.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further details, but embodiment does not limit in any form the present invention.Unless stated otherwise, the present invention adopts reagent, method and apparatus are the art conventional reagent, method and apparatus.
embodiment 1
Utilize a method for Acinetobacter bauamnnii synthetic flocculant, comprise the steps:
S1. actication of culture: by Acinetobacter bauamnnii
acinetobacterbaumanniiyNWH226 is inoculated in LB solid medium, activation culture 24h;
S2. cultivate: the single colony inoculation fresh activation obtained, in Congo red simulated wastewater, is cultivated 48h and obtained bacterium liquid;
S3. bacterium liquid S2 obtained in the centrifugal 30min of 8000g, go precipitation, get centrifugal after supernatant liquor mix 2h with ethanol in the ratio of 1:2;
S4. get the precipitated product that S3 obtains and carry out lyophilize 12h below-20 DEG C, obtain flocculation agent;
Wherein, Acinetobacter bauamnnii described in S1
acinetobacterbaumanniithe preserving number of YNWH226 is CGMCC7922;
Described in S2, Congo red simulated wastewater comprises: K
2hPO
41.6g/L, KH
2pO
40.2g/L, (NH
4)
2sO
41.0g/L, FeSO
47H
2o0.01g/L, NaCl0.1g/L, CaCl
26H
2o0.02g/L, MgSO
40.1g/L, glucose 1g/L, pH7.0, Congo red to 100mg/L after adding filtration sterilization after sterilizing in above-mentioned substratum.
Flocculation agent effect experimental: the dyeing waste water adopting Dongguan textile mills aeration tank, natural sedimentation 24h, removes supernatant liquor, gets the mud after 500mL precipitation to beaker, pH is adjusted to 6.0, adds the CaCl of the 16g/L of 5mL
2, add the flocculation agent of the above-mentioned synthesis of 1.3% of mud dry weight, 100rpm stirs 30min.Recording flocculating rate is 68%, and capillary water absorbing time (CST), resistivity are respectively by original 82.4s, 4.06 × 10
9s
2/ g becomes 63.4s, 2.93 × 10
9s
2/ g.Indices shows, after adding the flocculation agent of synthesis, sludge flocculation and dewatering obtain larger improvement, and effective pH scope is 5 ~ 8.
comparative example 1
The method that this comparative example and embodiment 1 prepare flocculation agent is substantially identical, and difference is, single colony inoculation that activation obtains by this comparative example S2 step, in LB nutrient solution, cultivates 48h.
Carry out effect experimental according to the method for embodiment 1 to above-mentioned obtained flocculation agent, recording flocculating rate is 62%, and capillary water absorbing time (CST), resistivity are respectively by original 82.4s, 4.06 × 10
9s
2/ g becomes 70.5s, 3.18 × 10
9s
2/ g.Indices shows, the flocculation agent made with Congo red simulated wastewater is than the flocculation agent better effects if made with LB nutrient solution, and cost is lower.
Claims (8)
1. utilize a method for Acinetobacter bauamnnii synthetic flocculant, it is characterized in that, comprise the steps:
S1. actication of culture: by Acinetobacter bauamnnii
acinetobacterbaumanniiyNWH226 is inoculated in solid medium and activates;
S2. cultivate: single colony inoculation activation obtained, in Congo red simulated wastewater, is cultivated and obtained bacterium liquid;
S3. the bacterium liquid obtained by S2 is centrifugal, get centrifugal after supernatant liquor mix with ethanol or polyoxyethylene glycol;
S4. get the precipitated product that S3 obtains and carry out lyophilize, obtain flocculation agent;
Wherein, Acinetobacter bauamnnii described in S1
acinetobacterbaumanniithe preserving number of YNWH226 is CGMCC7922;
Described in S2, Congo red simulated wastewater comprises: K
2hPO
41.6g/L, KH
2pO
40.2g/L, (NH
4)
2sO
41.0g/L, FeSO
47H
2o0.01g/L, NaCl0.1g/L, CaCl
26H
2o0.02g/L, MgSO
40.1g/L, glucose 1g/L, pH7.0, Congo red to 100mg/L after adding filtration sterilization after sterilizing in above-mentioned substratum.
2. method according to claim 1, is characterized in that, described in S1, solid medium comprises: bacteriological peptone 10g/L, yeast powder 5g/L, NaCl10g/L, agar powder 15g/L, pH7.0.
3. method according to claim 2, is characterized in that, the time activated described in S1 is 24h.
4. method according to claim 1, is characterized in that, the time of cultivating described in S2 is 48h.
5. method according to claim 1, is characterized in that, centrifugal described in S3 is the centrifugal 30min of 8000g.
6. method according to claim 1, is characterized in that, the volume ratio hybrid reaction 2h of 1:2 pressed by supernatant liquor described in S3 and ethanol.
7. method according to claim 1, is characterized in that, cryodesiccated condition described in S4 is-20 DEG C or following lyophilize 12h.
8. method according to claim 7, is characterized in that, described in S4, cryodesiccated temperature is-32 DEG C.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106947714A (en) * | 2017-03-17 | 2017-07-14 | 上海申启生物科技有限公司 | Acinetobacter bauamnnii selective enrichment broth and preparation method thereof |
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|---|---|---|---|---|
| CN102311975A (en) * | 2010-08-06 | 2012-01-11 | 北京联合大学 | Microorganism flocculating agent and preparation method thereof |
| CN102363793A (en) * | 2011-10-25 | 2012-02-29 | 杭州江南科学研究院有限公司 | Preparation method of photosynthetic bacteria flocculant |
| CN103756925A (en) * | 2013-10-22 | 2014-04-30 | 广东工业大学 | Acinetobacter baumannii, and screening method and application thereof in degradation of azo dye Congo red |
-
2015
- 2015-06-25 CN CN201510355798.6A patent/CN105039415B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102311975A (en) * | 2010-08-06 | 2012-01-11 | 北京联合大学 | Microorganism flocculating agent and preparation method thereof |
| CN102363793A (en) * | 2011-10-25 | 2012-02-29 | 杭州江南科学研究院有限公司 | Preparation method of photosynthetic bacteria flocculant |
| CN103756925A (en) * | 2013-10-22 | 2014-04-30 | 广东工业大学 | Acinetobacter baumannii, and screening method and application thereof in degradation of azo dye Congo red |
Non-Patent Citations (1)
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106947714A (en) * | 2017-03-17 | 2017-07-14 | 上海申启生物科技有限公司 | Acinetobacter bauamnnii selective enrichment broth and preparation method thereof |
| CN106947714B (en) * | 2017-03-17 | 2020-09-04 | 上海申启生物科技有限公司 | Acinetobacter baumannii selective enrichment liquid and preparation method thereof |
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|---|---|
| CN105039415B (en) | 2018-11-09 |
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Effective date of registration: 20230810 Address after: Dapingzi, Guanguang Industrial Zone, Guandu Town, Wengyuan County, Shaoguan City, Guangdong Province, 512000 Patentee after: SHAOGUAN PENGRUI ENVIRONMENTAL PROTECTION TECHNOLOGY Co.,Ltd. Address before: 510006 No. 100 West Ring Road, Guangzhou University, Guangzhou, Guangdong, Panyu District Patentee before: GUANGDONG University OF TECHNOLOGY |
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Address after: Dapingzi, Guanguang Industrial Zone, Guandu Town, Wengyuan County, Shaoguan City, Guangdong Province, 512000 Patentee after: Guangdong Pengrui Environmental Resources Co.,Ltd. Country or region after: China Address before: Dapingzi, Guanguang Industrial Zone, Guandu Town, Wengyuan County, Shaoguan City, Guangdong Province, 512000 Patentee before: SHAOGUAN PENGRUI ENVIRONMENTAL PROTECTION TECHNOLOGY Co.,Ltd. Country or region before: China |