CN107699516A - Halococci and its preparation and application - Google Patents

Halococci and its preparation and application Download PDF

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Publication number
CN107699516A
CN107699516A CN201710930440.0A CN201710930440A CN107699516A CN 107699516 A CN107699516 A CN 107699516A CN 201710930440 A CN201710930440 A CN 201710930440A CN 107699516 A CN107699516 A CN 107699516A
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China
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high salt
waste water
halococci
organic waste
preparation
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CN201710930440.0A
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Chinese (zh)
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陈标
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Jiangsu Luye Agrochemicals Co ltd
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Jiangsu Luye Agrochemicals Co ltd
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    • 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
    • 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
    • C12R2001/07—Bacillus
    • C12R2001/10—Bacillus licheniformis
    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/30—Organic compounds

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

Abstract

The invention discloses a kind of halococci and its preparation, it is CCTCC M 2017529 in the preserving number of China typical culture collection center, preservation date：On September 22nd, 2017, the entitled bacillus licheniformis GD5 LA of preservation.Preparation containing halococci is to use concentration as 108~109The high salt LB nutrient solutions of cfu/mL halococcis are lyophilized to be formed.The present invention screens by many experiments, obtains halophilic halococci, and carrying out high-salt wastewater processing using the bacterial strain of the present invention has the advantages of simple to operate, cost is low, degradation efficiency is high, has important application value to environmental protection.The bacterial strain is suitable for the environment of the high organic matter of high salt in waste water, has the advantages that the scope of application is wide and biological stability is good, bacterial strain of the invention has a good application prospect in terms of the processing of organic high-salinity wastewater.

Description

Halococci and its preparation and application
Technical field
The invention belongs to environmental technology field, more particularly to one plant of halococci and preparation containing the halococci and its The application of degradation of organic substances in high-salt wastewater.
Background technology
With the fast development of industry and the scarcity of water resource, industrial wastewater problem how is resolved just by more and more Concern.In industrial wastewater is greatly high salt organic waste water, such as dyestuff processing waste water, petroleum production wastewater and food Processing waste water etc..High-salt wastewater refers generally to total saliferous mass fraction >=1%, and salts substances contained therein are mostly Na+, Ca2+、 SO42-、 Cl-.Organic wastewater refers to organic concentration (in terms of COD) typically in more than 2000mg/L, and every liter had is even as high as It is tens of thousands of to ten tens of thousands of milligrams, and BOD5/COD values are less than 0.3 waste water.
At present, the processing method used both at home and abroad for high-salt wastewater mainly has Physical, chemical method and biological treatment. The method of traditional high salt organic waste water of processing mainly uses the physical and chemical process of " coagulation-filtering ", but the technique for The removal effect of most of dissolved organic matter such as phenols and its derivative is poor.Also, this method typically also needs to biological place Reason method carries out after-treatment to reach the discharge standard of industrial wastewater.
Biological treatment is a kind of most widely used method in current organic wastewater, mainly including biomembrance process, activity Sludge, Aerobic-anaerobic method etc..This method be mainly using microorganism itself metabolism, by adsorb, condense, aoxidize divide The organic matter that comes in degrading waste water of the effect such as solution, has that treating capacity is big, has a wide range of application, low cost and other advantages.At present, on dirt The research report of the degradation bacteria of Organic substance in water is a lot, but the degradation bacteria that organic wastewater can be handled under high salt conditions is less. Therefore, screening becomes particularly important suitable for the microorganism of efficient degradation organic matter in high-salt wastewater.
In order to solve the above problems, the invention provides it is a kind of can under the conditions of high salinity degradation of organic substances thermophilic salt ball Bacterium.
The content of the invention
It is an object of the invention to can efficiently be dropped in high-salt wastewater in view of the deficienciess of the prior art, providing one kind Solve the halococci of organic pollution, and application of the bacterial strain in high salt organic waste water is handled.
Technical scheme：To realize object above, the technical scheme that the present invention takes is:
A kind of halococci, it is CCTCC NO in the preserving number of China typical culture collection center:M 2017529, Preservation date：On September 22nd, 2017, the entitled bacillus licheniformis Bacillus licheniformisGD5-LA of preservation Clothing bacillus GD5-LA.Preservation address is Wuhan, China Wuhan University.
Another aspect of the present invention provides a kind of halococci preparation for high-salt wastewater processing, the halococci preparation In containing the present invention preserving number be CCTCC M 2017529 bacillus licheniformis GD5-LA.The halococci preparation includes Halococci nutrient solution freeze-dried powder, applied and preservation with facilitating.
The halococci preparation is 10 preferably with concentration8~109The high salt LB nutrient solutions of cfu/mL halococci freeze Dry powder is made.
Salinity in the high salt LB nutrient solutions is 20~120g/L, preferably 40~100g/L, concretely 100g/ L NaCl。
In the high salt LB culture mediums in addition to the NaCl for forming salinity, the cellar culture of bacterial growth is primarily to facilitate Base.The present invention using LB culture mediums, including：NaCl 10g, peptone 10g and yeast extract 5g.By each component 1L is settled to after dissolving.
It is preferred that the halococci is seeded in the high salt LB culture mediums, shaken in temperature for 25~40 DEG C (such as 37 DEG C) 12~48h of culture (such as 24h) is swung, then freezes nutrient solution.
Preferably, initial concentration of the halococci in inoculation can be 104~106Cfu/mL, such as 105cfu/mL.Shake Swing after culture terminates, the concentration of halococci can reach 10 described in the nutrient solution8~109cfu/mL.The concussion and cultivate Rotating speed be 100~250r/min, such as 220r/min.
Another aspect of the present invention provides the halococci and the above-mentioned preparation containing halococci of the present invention Application in high-salt wastewater processing.During using the halococci or preparation processing high-salt wastewater, the pH value of system can be 6.0~8.0, preferably 7.0~7.5.
Salinity in the high-salt wastewater can be more than 10g/L, can reach 20~120g/L.The salinity with NaCl is represented.
Another aspect provides a kind of processing method of high-salt wastewater, including：Institute is added in high-salt wastewater Halococci or the preparation are stated, and is fermented a period of time.The salinity of wherein described high-salt wastewater by NaCl be provided as 10g/L with On, preferably 20~120g/L, pH value is 6.0~8.0, preferably 7.0~7.5.
According to a kind of embodiment, the fermentation temperature of methods described is 15~40 DEG C, preferably 25~37 DEG C.Fermentation time For 60~80h.
The halococci put into according to a kind of embodiment in high-salt wastewater or the amount of the preparation are to make in waste water Initial bacteria concentration is 104~106cfu/mL。
The present invention is separated to organic matter in one plant of high-salt wastewater that can degrade from the sludge in local sewage treatment plant Bacterial strain.Through strain morphology, cultural character and molecular biology identification, it is halococci to determine the bacterium.The bacterial strain is to high-salt wastewater In organic matter there is good degradation effect.
The present invention has advantages below and effect：
(1) using the bacterial strain of the present invention carry out high-salt wastewater processing with it is simple to operate, that cost is low, degradation efficiency is high is excellent Point, there is important application value to environmental protection.
(2) bacterial strain is suitable for the environment of the high organic matter of high salt in waste water, has the scope of application wide good with biological stability The features such as.The bacterial strain of the present invention has a good application prospect in terms of the processing of organic high-salinity wastewater.
Embodiment
With reference to specific embodiment, the present invention is furture elucidated, it should be understood that these embodiments be merely to illustrate the present invention and Limitation the scope of the present invention is not used in, after the present invention has been read, various of equal value shapes of the those skilled in the art to the present invention The modification of formula falls within the application appended claims limited range.
The acquisition and identification of the halococci of embodiment 1
1st, the screening of halococci
Mud sample is derived from Wuxi City sludge sewage.
In 250mL triangular flasks, the LB fluid nutrient mediums of 1g mud samples and salinity for 40g/L (in terms of NaCl) are added 20mL, 1mL nutrient solutions are taken after 30 DEG C of isothermal vibration 24h in horizontal shaker, be added to the LB liquid that fresh salinity is 40g/L Continue to cultivate 24h in body culture medium, after so transferring 3 times repeatedly, step up the salinity in LB culture mediums, most LB liquid at last Salinity in culture medium is increased to 120g/L, and salinity is coated with as 120g/L's after then taking the nutrient solution gradient dilution under the salinity LB solid plates.Then, picking monoclonal bacterial strain carries out the degradation experiment of organic matter in high-salt wastewater from flat board, therefrom screens The bacterial strain strong to organic pollutant degradation ability.
2nd, the identification of halococci
Foundation《Common bacteria system identification handbook》, to the bacterial strain that separates of the present invention from individual morphology feature, colony characteristicses, Biochemical reactions etc. are identified, have then carried out 16S rRNA identifications to the bacterial strain again.Use primer 2 7F and 1492R with The strain gene group is that template enters performing PCR amplification, and amplified production is served into Hai Shenggong bioengineering limited company is carried out DNA sequencing, sequencing result submit NCBI to carry out BLAST retrievals, and it is bacillus licheniformis to determine the bacterial strain.
3rd, the preservation of halococci
It is CCTCC M 2017529 in the preserving number of China typical culture collection center, preservation date：2017 9 The moon 22, the entitled bacillus licheniformis GD5-LA of preservation.
Embodiment 2, the preparation of halococci preparation and its degraded to organic matter in high-salt wastewater
1st, the preparation of halococci preparation
Embodiment 1 is screened to obtained halococci and is seeded to the minimal medium that salinity (in terms of NaCl) is 120g/L In (pH 7.0), it is 10 to make its initial concentration5Cfu/mL, 37 DEG C, 220r/min concussion and cultivate 24h, culture terminate rear nutrient solution In halococci concentration be 108Cfu/mL, then the nutrient solution is freezed, obtain halococci preparation.
2nd, degraded of the halococci preparation to organic matter in high salt sanitary wastewater
By 1g, the halococci preparation is added to the life that 20mL salinity is 120g/L, initial organic concentration is 90mg/L In waste water (pH 8.5), 37 DEG C, 220r/min concussion and cultivates 60h.Then organic concentration in waste water is detected.Work as concussion and cultivate After 60h, the organic concentration in cultivating system is reduced to 41mg/L.The halococci preparation that the present invention is prepared has fine The degradation for high salinity organic matter, the feature of environmental protection of the present invention is strong, has important application value.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should It is considered as protection scope of the present invention.

Claims (10)

1. a kind of halococci, it is characterised in that it is CCTCC in the preserving number of China typical culture collection center M2017529, preservation date：On September 22nd, 2017.
2. a kind of preparation for being used to handle high salt organic waste water, it is characterised in that the preparation includes thermophilic described in claim 1 The freeze-dried powder of salt coccus nutrient solution.
3. the preparation according to claim 2 for being used to handle high salt organic waste water, it is characterised in that freeze-dried powder is to use concentration For 108~109The high salt LB nutrient solutions of cfu/mL halococcis are lyophilized to be formed.
4. the preparation according to claim 3 for being used to handle high salt organic waste water, it is characterised in that freeze-dried powder is by thermophilic salt Coccus is seeded in high salt LB culture mediums, in 25~40 DEG C of 12~48h of concussion and cultivate, is then freezed gained nutrient solution；Wherein The salinity of the high salt LB NaCl in medium is 20~120g/L.
5. application of the halococci in high salt organic waste water is handled described in claim 1.
6. the preparation for being used to handle high salt organic waste water described in any one of claim 2~4 is in high salt organic waste water is handled Application.
7. a kind of processing method of high salt organic waste water, it is characterised in that comprise the following steps：Put into high salt organic waste water The preparation described in halococci or input any one of claim 2~4 described in claim 1, and fermented and cultured is for a period of time.
8. the processing method of high salt organic waste water according to claim 7, it is characterised in that wherein described high salt organic waste The salt content of water, it is more than 10g/L in terms of NaCl, it is 6.0~8.0, preferably 7.0~7.5 to adjust organic wastewater pH.
9. the processing method of high salt organic waste water according to claim 7, it is characterised in that fermented and cultured temperature be 15~ 40 DEG C, preferably 25~37 DEG C, fermentation time are 60~80h.
10. the processing method of high salt organic waste water according to claim 7, it is characterised in that wherein described halococci Or the input amount of preparation is to make initial bacteria concentration in waste water be 104~106cfu/mL。
CN201710930440.0A 2017-10-09 2017-10-09 Halococci and its preparation and application Pending CN107699516A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114656043A (en) * 2022-04-01 2022-06-24 北京赛富威环境工程技术有限公司 Treatment method of high-salt low-concentration organic wastewater

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104894015A (en) * 2015-05-20 2015-09-09 北京维奥思环境工程有限公司 Bacillus and application thereof to degrade organic matter in high-salinity wastewater
CN105441359A (en) * 2015-12-17 2016-03-30 华南理工大学 Strain of bacillus licheniformis and applications thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104894015A (en) * 2015-05-20 2015-09-09 北京维奥思环境工程有限公司 Bacillus and application thereof to degrade organic matter in high-salinity wastewater
CN105441359A (en) * 2015-12-17 2016-03-30 华南理工大学 Strain of bacillus licheniformis and applications thereof

Non-Patent Citations (2)

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Title
TIAN LI ET AL.: "Biodegradation of Nitrobenzene in a Lysogeny Broth Medium by a Novel Halophilic Bacterium Bacillus Licheniformis", 《MARINE POLLUTION BULLETIN》 *
李昕等: "一株抗高温耐盐噬油菌处理含油废水的性能研究", 《环境工程》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114656043A (en) * 2022-04-01 2022-06-24 北京赛富威环境工程技术有限公司 Treatment method of high-salt low-concentration organic wastewater

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Application publication date: 20180216