CN105420170A - Aquatic comamonas and culture method and application thereof - Google Patents

Aquatic comamonas and culture method and application thereof Download PDF

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Publication number
CN105420170A
CN105420170A CN201511033629.7A CN201511033629A CN105420170A CN 105420170 A CN105420170 A CN 105420170A CN 201511033629 A CN201511033629 A CN 201511033629A CN 105420170 A CN105420170 A CN 105420170A
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comamonas
sewage
aquatic
aquatic comamonas
content
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CN105420170B (en
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王晓阳
李玉庆
张建
薛二军
韩超
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TIANJIN CARING TECHNOLOGY DEVELOPMENT CO LTD
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TIANJIN CARING TECHNOLOGY DEVELOPMENT CO LTD
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • 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
    • C12N1/205—Bacterial isolates
    • 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
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/26—Nature of the water, waste water, sewage or sludge to be treated from the processing of plants or parts thereof
    • C02F2103/28—Nature of the water, waste water, sewage or sludge to be treated from the processing of plants or parts thereof from the paper or cellulose industry

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Water Supply & Treatment (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Environmental & Geological Engineering (AREA)
  • Hydrology & Water Resources (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Paper (AREA)

Abstract

The invention provides aquatic comamonas and a culture method and application thereof. The aquatic comamonas is KY-02, and the preservation number is CGMCC No.11866. The aquatic comamonas KY-02 can degrade cellulose with the content of 1500 mg/L in sewage, so that the COD of sewage is lowered, the degradation onset time is short, and the degradation rate can reach about 15% on the second day. Compared with physical and chemical methods, the aquatic comamonas has the advantages of being low in cost, free of secondary pollution and the like, and has the wide application prospect and the good economic value in the field of papermaking sewage treatment.

Description

A kind of aquatic comamonas and cultural method thereof and application
Technical field
The present invention relates to microorganism and biological degradation field, particularly, relate to a kind of aquatic comamonas and cultural method thereof and application.
Background technology
Paper industry quantity of wastewater effluent is large, again containing a large amount of Mierocrystalline cellulose, Mierocrystalline cellulose, mineral alkali and resin, tannin, protein etc. in sewage, cause that Colority of Sewage Water is dark, basicity is large, hardly degraded organic substance content is high, oxygen-consumption large, belongs to typical comprehensive high concentration organic sewage.The industrial pollution caused by paper manufacturing of China is particularly serious.Paper industry, to the pollution serious threat environmental safety of water surrounding and the healthy of the mankind, is worldwide subject to extensive concern, and the pollution solving papermaking wastewater becomes the task of top priority.
In recent decades, people have carried out quite detailed research to the biological degradation of pollutent, and achieve many achievements.But microorganism is relatively less in the biological degradation research of Industry Waste water pollutant, and the condition that mainly trade effluent is severe restricted, as low in high salinity, extremely pH, dissolved oxygen amount etc.Common micro-organisms and active sludge are all difficult to realize cellulosic effective degraded.Focus mostly in chemical oxidation methods such as Fenton reagent factures for treatment process cellulosic in papermaking wastewater at present.But chemical process needs to add a large amount of medicaments, and cost is higher, also easily causes secondary pollution.Therefore, by screening degradation bacteria, the Mierocrystalline cellulose in disposing of sewage with biological process becomes optimal selection.
Comamonas belongs to 1985 by foundation such as DeVos, and the kind being attributed to this genus at present comprises Comamonas terrigena, Comamonas testosteroni, Comamonas acidovorans, aquatic comamonas etc.This bacterioid for the pollutents such as phenols, quinoline class degraded very early studied personnel find, due to the otherness of genetic background, the pollutent that different comamonas can be degraded and degradation characteristic all different.
Summary of the invention
The object of the invention is to make up defect of the prior art, provide a kind of can Mierocrystalline cellulose in degradation of sewage under aerobic condition, thus reduce the COD of sewage, and degraded onset time short aquatic comamonas and cultural method and application.
Aquatic comamonas KY-02 of the present invention screens by pulping papermaking sewerage factory active sludge, its bacterium colony lawn in the form of sheets on culture plate, lawn is flat opaque, very large, white, surperficial fine hair shape is dry, edge is that branch splits, monomer is thick bar shape, Gram-negative, has stronger soluble cellulose degradation capability.Based on morphological specificity, physiological and biochemical property, be aquatic comamonas by this identification of strains, called after KY-02.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 11st, 2015 and (is called for short CGMCC, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), deposit number is CGMCCNo.11866, and classification is by name aquatic from hair Zymomonas mobilis Comamonasaquatic.
The invention provides and cultivate the above-mentioned substratum stating aquatic comamonas KY-02, its formula is, in every 1L water, contain: (NH 4) 2sO 42g, K 2hPO40.5g, NaCl0.5g, MgSO 40.5g, KNO 31g, Mierocrystalline cellulose sulfonate 0.5-1.5g, pH7.2, solid-state add 2% agar.
Above-mentioned substratum, is prepared by following method: make substratum for subsequent use by water dissolution after above-mentioned each component weighs by prescription, in 121 DEG C, and sterilizing 20-30min under 102kpa.
Microbial inoculum containing described aquatic comamonas KY-02 or biological cleanser belong to protection scope of the present invention.
Further, in above-mentioned microbial inoculum or biological cleanser, the content of aquatic comamonas KY-02 is 10 5-10 8/ ml.Control moisture at 15%-20%.
The invention provides aquatic comamonas KY-02 or containing the microbial inoculum of this bacterium or the application of biological cleanser in cleaning ambient.
The invention provides aquatic comamonas KY-02 or containing the microbial inoculum of this bacterium or the application of biological cleanser in purified industrial waste water.
The invention provides aquatic comamonas KY-02 or the microbial inoculum containing this bacterium or the biological cleanser application in degraded papermaking wastewater in content of cellulose.
Further, above-mentioned application is aquatic comamonas KY-02 concentration is 10 5-10 6during/ml, drop in sewage with the inoculum size of 5%-15%.
Wherein, the fiber cellulose content≤1500mg/L in described sewage.
Preferably, content of cellulose 200-570mg/L in described sewage.
More preferably, content of cellulose 250-570mg/L in described sewage.
Aquatic comamonas KY-02 can Mierocrystalline cellulose in degradation of sewage under aerobic condition, and the content of cellulose in degradable sewage is 1500mg/L, thus reduces the COD of sewage, and onset time of degrading is short, just can reach about 15% at the 2nd day degradation rate.There is the advantages such as cost is low, non-secondary pollution compared with physics, chemical process, have wide practical use in Sewage field and good economic worth.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, the amendment do the inventive method, step or condition or replacement, all belong to scope of the present invention.
If do not specialize, chemical reagent used in embodiment is conventional commercial reagent, the conventional means that technique means used in embodiment is well known to those skilled in the art.
The separation andpreconcentration of the aquatic comamonas KY-02 of embodiment 1
Aquatic comamonas KY-02 of the present invention screens by pulping papermaking sewerage factory active sludge, its bacterium colony lawn in the form of sheets on culture plate, lawn is flat opaque, very large, white, surperficial fine hair shape is dry, edge is that branch splits, monomer is thick bar shape, Gram-negative, has stronger soluble cellulose degradation capability.
This identification of strains is aquatic comamonas by comprehensive thalli morphology, physio-biochemical characteristics and 16SrDNA gene order (as shown in SEQIDNO.1), called after KY-02.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 11st, 2015 and (is called for short CGMCC, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), deposit number is CGMCCNo.11866, and classification is by name aquatic from hair Zymomonas mobilis Comamonasaquatic.
The performance test of the degraded cellulose of the aquatic comamonas KY-02 of embodiment 2
Experiment one, described aquatic comamonas KY-02 accessed in substratum and cultivates, wherein the consisting of of substratum: (NH 4) 2sO 42g, K 2hPO40.5g, NaCl0.5g, MgSO 40.5g, KNO 31g, Mierocrystalline cellulose sulfonate 0.5-1.5g, distilled water 1000ml, PH7.2 cultivate and are about 10 to bacterium is dense in 2 days 5-10 6/ ml; With the inoculum size of 5%, the aerobic section water outlet of Tianjin Milky Way Zhi Ye Sewage factory and Ninghe Zhi Ye Sewage factory is experimental subjects, often will locate sewage 100ml loads in 500ml triangular flask (being numbered 1 and 2), triangle bottleneck is sealed with silica gel plug, at 180r/min, cultivate 2 days at 30 DEG C.Sample respectively, centrifugal rear ultraviolet spectrophotometry surveys content of cellulose in supernatant liquor.The results are shown in Table 1.
Experiment two, the substratum content of use and the amount of inoculation are different with embodiment 2, sewage numbering 3 and 4
Implementation method: described aquatic comamonas KY-02 is accessed in substratum and cultivates, wherein the consisting of of substratum: (NH 4) 2sO 42g, K 2hPO40.5g, NaCl0.5g, MgSO 40.5g, KNO 31g, Mierocrystalline cellulose sulfonate 0.5-1.5g, distilled water 1000ml, PH7.2 cultivate and are about 10 to bacterium is dense in 2 days 5-10 6/ ml; With the inoculum size of 10%, the aerobic section water outlet of Tianjin Milky Way Zhi Ye Sewage factory and Ninghe Zhi Ye Sewage factory is experimental subjects, often will locate sewage 100ml loads in 500ml triangular flask (being numbered 3 and 4), triangle bottleneck is sealed with silica gel plug, at 180r/min, cultivate 2 days at 30 DEG C.Sample respectively, centrifugal rear ultraviolet spectrophotometry surveys content of cellulose in supernatant liquor.The results are shown in Table 1.
Experiment three, the substratum content of use and the amount of inoculation are different with embodiment 2 and 3, sewage numbering 5 and 6
Implementation method: described aquatic comamonas KY-02 is accessed in substratum and cultivates, wherein the consisting of of substratum: (NH 4) 2sO 42g, K 2hPO40.5g, NaCl0.5g, MgSO 40.5g, KNO 31g, Mierocrystalline cellulose sulfonate 0.5-1.5g, heats up in a steamer water 1000ml, and PH7.2 cultivates and is about 10 to bacterium is dense in 2 days 5-10 6/ ml; With the inoculum size of 15%, the aerobic section water outlet of Tianjin Milky Way Zhi Ye Sewage factory and Ninghe Zhi Ye Sewage factory is experimental subjects, often will locate sewage 100ml loads in 500ml triangular flask (being numbered 5 and 6), triangle bottleneck is sealed with silica gel plug, at 180r/min, cultivate 2 days at 30 DEG C.Sample respectively, centrifugal rear ultraviolet spectrophotometry surveys content of cellulose in supernatant liquor.The results are shown in Table 1.
Table 1
Cellulosic measuring method: compound concentration is 20,40 respectively, the Mierocrystalline cellulose sulfonate solution of 60,40,100,120 (mg/L).Use ultraviolet spectrophotometry, adjusting wavelength is 276nm, surveys its absorbancy.Then according to measuring result drawing standard curve.Get sewage sample 5ml, centrifugal 10min under 5000rpm.Get supernatant liquor and be diluted to 5-10 doubly, make content of cellulose within the scope of 20-120mg/L.Use ultraviolet spectrophotometry, the light absorption value of working sample, then according to typical curve and extension rate, calculate cellulosic content in sample.
The method of calculation of degradation rate: content of cellulose before degradation rate=(before process the rear content of cellulose of content of cellulose-process)/process
Result shows: aquatic comamonas KY-02 have significant degradation capability for the fiber in Milky Way paper industry and Ninghe paper industry paper waste, even if content of cellulose is up to 571mg/L in sewage, degradation rate also can reach about 16%, and degradation effect is stablized, repeatedly, degradation effect is basically identical.
Embodiment 3 aquatic comamonas KY-02 degraded COD
Implementation method:
1, described aquatic comamonas KY-02 is seeded to LB substratum, 30 DEG C of shaking culture are about 10 to bacterium is dense 7-10 8/ ml;
2, be inoculated in 50L aeration tank with 10% inoculum size, under room temperature (22-27 DEG C), aeration runs 4 days, and maintenance dissolved oxygen is 4ppm;
3, system stops aeration, and precipitation gets supernatant, measures COD according to " dichromate titration (GB11914-89) ".
Handling object is the aerobic section water outlet of different Sewage factory of two places in embodiment 2-4, and be numbered 1 and 2 respectively, principal pollutant are Mierocrystalline cellulose and degraded product thereof.Result is as table 2.
Table 2
The method of calculation of degradation rate: COD before degradation rate=(before process the rear COD of COD-process)/process
Result shows: aquatic comamonas KY-02 can effectively degrade the COD of Milky Way paper industry and peace paper industry papermaking wastewater, its effect and to remove cellulosic effect consistent.

Claims (10)

1. aquatic comamonas (Comamonasaquatic) KY-02, its deposit number is CGMCCNo.11866.
2. cultivate the substratum of aquatic comamonas KY-02 described in claim 1, its formula is, in every 1L water, contain: (NH 4) 2sO 42g, K 2hPO40.5g, NaCl0.5g, MgSO 40.5g, KNO 31g, Mierocrystalline cellulose sulfonate 0.5-1.5g, pH7.2, solid-state add 2% agar.
3. the microbial inoculum containing aquatic comamonas KY-02 described in claim 1 or biological cleanser.
4. microbial inoculum as claimed in claim 3 or biological cleanser, it is characterized in that, the content of aquatic comamonas KY-02 is 10 5-10 8/ ml.
5. aquatic comamonas KY-02 according to claim 1 or microbial inoculum according to claim 3 or the application of biological cleanser in cleaning ambient.
6. aquatic comamonas KY-02 according to claim 1 or microbial inoculum according to claim 3 or the application of biological cleanser in purified industrial waste water.
7. aquatic comamonas KY-02 according to claim 1 or microbial inoculum according to claim 3 or the application of biological cleanser in degraded papermaking wastewater in content of cellulose.
8. apply as claimed in claim 7, it is characterized in that, aquatic comamonas KY-02 concentration is 10 5-10 6during/ml, drop in sewage with the inoculum size of 5-15%.
9. apply as claimed in claim 7 or 8, it is characterized in that, the content of cellulose≤1500mg/L in sewage.
10. apply as claimed in claim 9, it is characterized in that, content of cellulose 200-570mg/L in sewage.
CN201511033629.7A 2015-12-30 2015-12-30 A kind of aquatic comamonas and its cultural method and application Active CN105420170B (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112920964A (en) * 2020-12-23 2021-06-08 南京振旭生物科技有限公司 Comamonas hydrocerussitum strain for preventing and treating tomato wilt, biocontrol microbial inoculum and preparation method and application thereof
CN115259397A (en) * 2022-06-30 2022-11-01 苏州鑫泷生态环境有限公司 Composite strain for treating aquaculture wastewater

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101348772A (en) * 2008-02-19 2009-01-21 南京大学 Comamonas aquatica LNL3 and use thereof in waste water biological denitrification
CN104450591A (en) * 2014-12-28 2015-03-25 天津凯英科技发展有限公司 Pseudoxanthomonas.Sp KYLI-01 and cultivation and application thereof
CN104630098A (en) * 2014-12-29 2015-05-20 天津凯英科技发展有限公司 Pseudomonas monteilii KY-05 and application thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101348772A (en) * 2008-02-19 2009-01-21 南京大学 Comamonas aquatica LNL3 and use thereof in waste water biological denitrification
CN104450591A (en) * 2014-12-28 2015-03-25 天津凯英科技发展有限公司 Pseudoxanthomonas.Sp KYLI-01 and cultivation and application thereof
CN104630098A (en) * 2014-12-29 2015-05-20 天津凯英科技发展有限公司 Pseudomonas monteilii KY-05 and application thereof

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112920964A (en) * 2020-12-23 2021-06-08 南京振旭生物科技有限公司 Comamonas hydrocerussitum strain for preventing and treating tomato wilt, biocontrol microbial inoculum and preparation method and application thereof
CN115259397A (en) * 2022-06-30 2022-11-01 苏州鑫泷生态环境有限公司 Composite strain for treating aquaculture wastewater

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