CN103275908B - Low-temperature denitrification pseudomonas fluorescens - Google Patents

Low-temperature denitrification pseudomonas fluorescens Download PDF

Info

Publication number
CN103275908B
CN103275908B CN201310248107.3A CN201310248107A CN103275908B CN 103275908 B CN103275908 B CN 103275908B CN 201310248107 A CN201310248107 A CN 201310248107A CN 103275908 B CN103275908 B CN 103275908B
Authority
CN
China
Prior art keywords
low
pseudomonas fluorescens
denitrification
temperature
nitrogen
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN201310248107.3A
Other languages
Chinese (zh)
Other versions
CN103275908A (en
Inventor
张淑梅
姜威
高娃
李晶
孟利强
赵晓宇
陈靖宇
曹旭
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Microbiology of Heilongjiang Academy of Sciences
Original Assignee
Institute of Microbiology of Heilongjiang Academy of Sciences
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute of Microbiology of Heilongjiang Academy of Sciences filed Critical Institute of Microbiology of Heilongjiang Academy of Sciences
Priority to CN201310248107.3A priority Critical patent/CN103275908B/en
Publication of CN103275908A publication Critical patent/CN103275908A/en
Application granted granted Critical
Publication of CN103275908B publication Critical patent/CN103275908B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses low-temperature denitrification pseudomonas fluorescens and relates to pseudomonas fluorescens. The low-temperature denitrification pseudomonas fluorescens are pseudomonas fluorescens WSW-1001 and are collected in China General Microbiological Culture Collection Center (CGMCC), the collection number is CGMCC No.7476, the collection date is April 15th, 2013, and the collection address is No.3, No.1 Courtyard, Beichen Road, Chaoyang District, Beijing. The low-temperature denitrification pseudomonas fluorescens are heterotrophic nitrification aerobic denitrifiers, can be used for degrading NH4+-N, NO--N and NO--N (0.25 mg/L-1314.1 mg/L) with a wide concentration range, can be applied to low-nitrogen pollution source water denitrification, medium- and high-concentration wastewater denitrification and eutrophic water body treatment, and have an important meaning on low-nitrogen pollution source water denitrification, water eutrophication control and biological environment modification.

Description

One strain low-temperature denitrification Pseudomonas fluorescence
Technical field
The present invention relates to a strain Pseudomonas fluorescence.
Background technology
Along with society and economic fast development, the polluted by nitrogen in China's water surrounding is on the rise.Sanitary sewage, trade effluent, breeding wastewater and farmland nitrogenous fertilizer are the main nitrogen pollution sources of water surrounding.Water body polluted by nitrogen not only causes serious environmental pollution and financial loss, even threatens human health and ecological safety.Microbial denitrogenation is considered at present most economical effective and the most promising water body and denitrogenates technology because having the advantages such as high-level efficiency, low cost, non-secondary pollution.
Nitrogen in polluted-water mainly comprises NH 4 +-N, NO 3 --N and NO 2 --N inorganic nitrogen and organonitrogen.Traditional biological denitrificaion comprises ammonification, nitrification and denitrification process, has been worked in coordination with by different oxygen, nitrification and denitrification bacterium, and the organonitrogen in sewage is ammonia nitrogen by heterotroph microbiological oxidation decomposition and inversion, is then NO by Autotrophic nitrification bacterium by mineralized nitrogen 3 --N and NO 2 --N, finally by denitrifying bacterium by NO 3 --N and NO 2 --N is converted into N 2.Recent study shows that some Heterotrophic nitrification aerobic denitrifying bacterias have synchronous nitration and denitrification function, can complete independently denitrogenation whole process.
Biological denitrificaion efficiency is limited by temperature and nitrogen concentration, and general bacterial strain is narrower to the subject range of nitrogen concentration, declines at low temperature (10 DEG C of <) denitrification ability, is not suitable for low-temperature water source denitrogenation of water.
Summary of the invention
The object of the invention is to provide a strain low-temperature denitrification Pseudomonas fluorescence.
One strain low-temperature denitrification Pseudomonas fluorescence, it is Pseudomonas fluorescens (Pseudomonasfluorescens) WSW-1001, in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, deposit number is CGMCC No.7476, preservation date is on April 15th, 2013, and preservation address is No. 3, No. 1, Beichen Lu, Chaoyang District, Beijing City institute.
Pseudomonas fluorescens (Pseudomonas fluorescens) WSW-1001, the morphological specificity of bacterial strain: it is rod-short, without gemma, size is 0.2~0.4 μ m × 1.0~1.3 μ m; Bacterium colony circle, smooth, neat in edge, on NYD substratum, bacterium colony is orange-yellow, in minimal media liquid, cultivates and can produce ceroid.
Pseudomonas fluorescens (Pseudomonas fluorescens) WSW-1001, the physiological and biochemical property of bacterial strain: it is Gram-negative aerobic bacteria, oxidase positive, the catalase positive, the arginine dihydrolase positive, utilizes dextrose plus saccharose, V.P feminine gender, M.R feminine gender, liquefy gelatin; Growth temperature is 4~37 DEG C, and optimum growth temperature is 15~20 DEG C, and growth pH value is 4.0~10.0, and the most suitable growth pH value is 7.0~8.0, C/N=1~20, the suitableeest C/N=10.
Pseudomonas fluorescens (Pseudomonas fluorescens) WSW-1001, the molecular biology identification result of bacterial strain: analyze by 16S rDNA sequence alignment, the most approaching with the 16S rDNA sequence of Pseudomonas fluorescence, similarity is 99%; In conjunction with morphological features, growth characteristics, physio-biochemical characteristics determine that bacterial strain WSW-1001 is Pseudomonas fluorescens (Pseudomonas fluorescens).
The present invention's one strain low-temperature denitrification Pseudomonas fluorescence, it is Pseudomonas fluorescens (Pseudomonasfluorescens) WSW-1001, in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, deposit number is CGMCC No.7476, preservation date is on April 15th, 2013, and preservation address is No. 3, No. 1, Beichen Lu, Chaoyang District, Beijing City institute.
Low-temperature denitrification Pseudomonas fluorescence in the present invention can be under 8 DEG C of conditions the NH of the low middle and high concentration in efficient degradation water 4 +-N, NO 3 --N and NO 2-N, the NH of 24h to low middle and high concentration 4 +-N (0.25-4.53mg/L, 48.34-365.32mg/L, 522.13-1314.24mg/L) degradation rate is 78.4-72.3%, 71.4%-50.7%, 23.2%-12.4%, to the NO of low middle and high concentration 3 -the degradation rate of-N (0.24-5.08mg/L, 50.41-352.35mg/L, 513.44-1201.8mg/L) is 83.3%-81.2%, 81.8%-66.9%, 40.5%-16.4%; To the NO of low middle and high concentration 2 -the degradation rate of-N (0.27-5.24mg/L, 49.83-308.92mg/L, 518.67-1084.45mg/L) is 84.4%-83.2%, 80.2%-76.5%, 34.5%-18.9%.
Low-temperature denitrification Pseudomonas fluorescence in the present invention is Heterotrophic nitrification aerobic denitrifying bacteria, can under 8 DEG C of conditions, synchronously carry out nitrated and aerobic anti-nitration reaction, can independently efficiently complete denitrogenation whole process.Initial ammonia nitrogen is 48.34mg/L, 8 DEG C, 24h can be that nitrogen is got rid of by 47.8% mineralized nitrogen, residual ammonia nitrogen 18.81 ± 0.15mg/L, there is a small amount of nitric nitrogen (5.04 ± 0.12mg/L), nitrite nitrogen (0.002 ± 0.001mg/L) and the N of trace 2o (0.003 ± 0.0013mg/L) produces, and approximately 20.9% nitrogen becomes cellular material.
Low-temperature denitrification Pseudomonas fluorescence in the present invention, the NH of the extensive concentration range of can degrading 4 +-N, NO 3 --N and NO 2 --N (0.25mg/L-1314.1mg/L), can be applicable to the source water denitrogenation of low polluted by nitrogen, denitrogenation of waste water and the eutrophication water processing of senior middle school's concentration, to low temperature (10 DEG C of <) source water denitrogenation, body eutrophication is administered and ecotope is repaired significant.
Brief description of the drawings
Fig. 1 is the colonial morphology figure of low-temperature denitrification Pseudomonas fluorescence on NYD substratum in embodiment one;
Fig. 2 is the phyletic evolution tree graph of low-temperature denitrification Pseudomonas fluorescence in embodiment one.
Embodiment
Embodiment one: present embodiment one strain low-temperature denitrification Pseudomonas fluorescence, it is Pseudomonas fluorescens (Pseudomonas fluorescens) WSW-1001, in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, deposit number is CGMCC No.7476, preservation date is on April 15th, 2013, and preservation address is No. 3, No. 1, Beichen Lu, Chaoyang District, Beijing City institute.
A strain low-temperature denitrification Pseudomonas fluorescence in present embodiment, its separation screening from Heilongjiang Province's Songhuajiang River Water in March obtains, screening process is as follows: get 100mL river and put into 500mL triangular flask, add 50mg ammonium sulfate, 300mg sodium-acetate and 5mL trace element solution, 8 DEG C, 160r/min enrichment culture 5 days, getting 1mL enrichment culture liquid adds in the triangular flask that contains 50mL basic culture solution, 3 Duplicate Samples, 8 DEG C, 160r/min concussion is cultivated 3 days, by nutrient solution doubling dilution to 1000 times, getting 100 μ L nutrient solutions is coated on the solid plate containing minimum medium, flat board is placed in 8 DEG C of incubators and is cultivated until grow single bacterium colony, to after single bacterium colony line purifying, carry out the test of low temperature ammonia nitrogen degradation, can obtain low temperature ammonia nitrogen degradation bacterial strain, wherein every 1L basic culture solution is made up of the distilled water of 0.5g ammonium sulfate, 5g sodium-acetate, 50mL trace element solution and surplus, 121 DEG C of sterilizing 20min, described trace element solution (g/L): phosphorus 5g acid hydrogen dipotassium, 2.5g magnesium sulfate, 2.5g sodium-chlor, 0.05g ferric sulfate, 0.05g manganous sulfate and 1L distilled water water, minimum medium is added with agar and is formed by basic culture solution.
Obtained bacterial strain is carried out to form, cultural characters and physio-biochemical characteristics qualification, and concrete steps are as follows: the thalline that low temperature is cultivated 48 hours carries out under 1000 power microscopes, observing thalline size and thalline color after gramstaining; To after bacterium liquid doubling dilution, be coated on minimum medium and NYD culture medium flat plate, observe colonial morphology and color; Liquid bacteria liquid is cultivated to 4-7 days, observe and have or not pigment formation; Bacterial strain is carried out to oxydase, catalase, arginine dihydrolase, dextrose plus saccharose fermentation, V.P, M.R and gelatin liquification test.
Result: this bacterial strain is rod-short, without gemma, size is 0.2~0.4 μ m × 1.0~1.3 μ m; Bacterium colony circle, smooth, neat in edge (as Fig. 1), on NYD substratum, bacterium colony is orange-yellow, in minimal media liquid, cultivates and can produce ceroid.This bacterial strain is Gram-negative aerobic bacteria, oxidase positive, and the catalase positive, the arginine dihydrolase positive, utilizes dextrose plus saccharose, V.P feminine gender, M.R feminine gender, liquefy gelatin; Growth temperature is 4~37 DEG C, and optimum growth temperature is 15~20 DEG C.
Obtained bacterial strain is carried out to 16S rDNA sequential analysis: get the centrifugal collection thalline of 1mL bacterium liquid, add 100 μ L sterilized waters, after suspension, boil 10min, centrifugal rear collection supernatant liquor obtains total DNA, carry out pcr amplification with bacterial 16 S rDNA universal primer, PCR product is carried out to agargel electrophoresis, reclaiming size checks order at the unique DNA band at 1500bp place, result obtains 1400bp DNA sequence dna, in GenBank, carry out homologous sequence comparison, with Mega4.0 software building phylogenetic tree (as Fig. 2), the 16S rDNA sequence homology 99% of result and Pseudomonas fluorescence, combining form, cultural characters and physio-biochemical characteristics, be accredited as Pseudomonas fluorescens (Pseudomonas fluorescens), therefore the bacterial strain that obtained is a novel bacterial, be committed to the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, deposit number is CGMCC No.7476, preservation date is on April 15th, 2013.
The Heterotrophic nitrification of low-temperature denitrification Pseudomonas fluorescence and aerobic denitrification:
The utilization of bacterial strain to different carbon sources: taking ammonium sulfate as only nitrogen source, change carbon source, carbon source is respectively citric acid, trisodium citrate, glucose, sucrose, sodium-acetate, sodium carbonate and glycerine.Ammonium sulfate content in nutrient solution is 0.05%, and carbon source content is 0.5%, trace element 5%, pH7.2.50mL nutrient solution is contained in 250mL triangular flask, and 8 DEG C, 160r/min concussion is cultivated 2 days, and 3 of each processing are parallel, measure bacterium turbidity (OD 600).The results are shown in Table 1, bacterial strain can finely utilize 7 kinds of carbon sources, Dichlorodiphenyl Acetate salt utilize effect best.
The impact of the different carbon sources of table 1 on strain growth
The utilization of bacterial strain to different nitrogen sources: taking sodium-acetate as sole carbon source, change nitrogenous source, nitrogenous source is respectively ammonium chloride, ammonium sulfate, lemon acid amide, azanol, saltpetre, Sodium Nitrite.Sodium acetate content in nutrient solution is 0.5%, and nitrogenous source content is 0.05%, trace element 5%, pH7.2.50ml nutrient solution is contained in 250ml triangular flask, and 8 DEG C, 160r/min concussion is cultivated 2 days, and 3 of each processing are parallel, measure bacterium turbidity (OD 600).The results are shown in Table 2, bacterial strain can finely utilize 6 kinds of nitrogenous sources.
The impact of table 2 different nitrogen sources on strain growth
The degradation capability of bacterial strain to different concns nitrogenous source: taking sodium-acetate as sole carbon source, nitrogenous source is used respectively ammonium sulfate, saltpetre and Sodium Nitrite, concentration is divided 3 sections, 0.2-6.0.mg/L, 50-400mg/L, 500-1000mg/L.50mL nutrient solution is contained in 250mL triangular flask, 8 DEG C, 160r/min concussion is cultivated 1 day, 3 of each processing are parallel, measure residual nitrogen content and (adopt nessler reagent light-intensity method according to GB7480-87 ammonia nitrogen, nitrite adopts N-Nai Ji-quadrol light-intensity method, and nitrate adopts Pbenoldisulfonic Acid light-intensity method).The results are shown in Table 3, the NH of the extensive concentration range of bacterial strain degradable 4 -n, NO 3 -n and NO 2 -n, to the degradation capability of high density nitrogenous source (> 500mg/L) lower than low middle concentration.
The degradation effect of table 3 bacterial strain to different concns nitrogenous source
Starting point concentration mg/L Final concentration mg/L Nitrogen degradation rate %
NH 4 --N 0.25±0.12 0.054±0.15 78.4
4.53±0.15 1.257±0.18 72.3
48.34±0.21 18.81±0.23 71.4
165.43±0.14 62.19±0.12 62.4
365.32±0.17 180.09±0.13 50.7
522.13±0.19 400.97±0.16 23.2
1314.24±0.16 1151.16±0.12 12.4
N0 3 --N 0.24±0.12 0.04±0.14 83.3
5.08±0.17 0.95±0.14 81.2
50.41±013 9.17±0.12 81.8
97.83±0.16 19.95±0.20 79.6
352.35±0.21 116.63±0.18 66.9
513.44±0.23 305.50±0.19 40.5
1201.80±0.14 1004.71±0.19 16.4
NO 2 --N 0.27±0.25 0.05±0.21 84.4
5.24±0.22 0.88±0.19 83.2
49.83±0.20 9.86±0.23 80.2
124.53±0.14 27.65±0.17 77.8
308.92±0.21 72.6±0.24 76.5
518.67±0.18 339.73±0.23 34.5
1084.45±0.26 879.49±0.20 18.9

Claims (1)

1. a strain low-temperature denitrification Pseudomonas fluorescence, it is characterized in that it is Pseudomonas fluorescens (Pseudomonas fluorescens) WSW-1001, in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, deposit number is CGMCC No.7476, preservation date is on April 15th, 2013, and preservation address is No. 3, No. 1, Beichen Lu, Chaoyang District, Beijing City institute.
CN201310248107.3A 2013-06-21 2013-06-21 Low-temperature denitrification pseudomonas fluorescens Expired - Fee Related CN103275908B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201310248107.3A CN103275908B (en) 2013-06-21 2013-06-21 Low-temperature denitrification pseudomonas fluorescens

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201310248107.3A CN103275908B (en) 2013-06-21 2013-06-21 Low-temperature denitrification pseudomonas fluorescens

Publications (2)

Publication Number Publication Date
CN103275908A CN103275908A (en) 2013-09-04
CN103275908B true CN103275908B (en) 2014-10-08

Family

ID=49058534

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201310248107.3A Expired - Fee Related CN103275908B (en) 2013-06-21 2013-06-21 Low-temperature denitrification pseudomonas fluorescens

Country Status (1)

Country Link
CN (1) CN103275908B (en)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104164389B (en) * 2014-07-09 2016-09-07 青岛蔚蓝天成生物科技有限公司 One fluorescent pseudomonads and the application in aquaculture thereof
CN109179650A (en) * 2018-09-21 2019-01-11 西安吉利电子化工有限公司 A kind of high-performance bio compounded carbons and preparation method thereof
CN112625942B (en) * 2020-12-01 2022-06-14 华南理工大学 Aerobic denitrifying bacterium and application thereof
CN113151077B (en) * 2021-04-14 2023-03-21 广西大学 Pseudomonas aeruginosa and screening method and application thereof
CN114058554B (en) * 2021-12-10 2022-09-13 青岛蔚蓝赛德生物科技有限公司 Composting pseudomonas strain and application thereof
CN114836355B (en) * 2022-05-30 2024-04-02 沈阳风景园林股份有限公司 Pseudomonas DT04 and application thereof in sewage denitrification

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102061276A (en) * 2010-11-15 2011-05-18 北京大学 Pseudomonas sp. strain for biological denitrification under low temperature and application thereof
CN102776140A (en) * 2012-05-17 2012-11-14 哈尔滨工业大学宜兴环保研究院 Cold-tolerant pseudomonas strain Den-05, and screening method and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102061276A (en) * 2010-11-15 2011-05-18 北京大学 Pseudomonas sp. strain for biological denitrification under low temperature and application thereof
CN102776140A (en) * 2012-05-17 2012-11-14 哈尔滨工业大学宜兴环保研究院 Cold-tolerant pseudomonas strain Den-05, and screening method and application thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
一株具有多途径氮代谢功能的荧光假单胞菌;王晋宇 等;《土壤学报》;20070131;第44卷(第1期);144-149页 *
同步脱氮除硫菌株的筛选、分离和鉴定;陈子爱;《中国沼气》;20081230;第26卷(第6期);3-7、12页 *
王晋宇 等.一株具有多途径氮代谢功能的荧光假单胞菌.《土壤学报》.2007,第44卷(第1期),144-149页.
陈子爱.同步脱氮除硫菌株的筛选、分离和鉴定.《中国沼气》.2008,第26卷(第6期),3-7、12页.

Also Published As

Publication number Publication date
CN103275908A (en) 2013-09-04

Similar Documents

Publication Publication Date Title
Zheng et al. Enhanced removal mechanism of iron carbon micro-electrolysis constructed wetland on C, N, and P in salty permitted effluent of wastewater treatment plant
Wen et al. Treatment of anaerobically digested swine wastewater by Rhodobacter blasticus and Rhodobacter capsulatus
Su et al. Comparison of nutrient removal capacity and biomass settleability of four high-potential microalgal species
CN103275908B (en) Low-temperature denitrification pseudomonas fluorescens
CN101570738B (en) Agrobacterium with heterotrophic nitrification-aerobic denitrification capability and application thereof in nitrogenous effluent treatment
CN102703350B (en) Application of salt-tolerant nitrogen and phosphorus removing bacillus altitudinis to wastewater treatment
CN102747014B (en) High-saline biological denitrification salinivibrio strain and application thereof in wastewater treatment
CN103497905B (en) A kind of efficient denitrification flcos producing bacteria strain and application thereof
CN103103153B (en) Anaerobic denitrification phosphorus-accumulating bacteria strain with denitrification and phosphorous removal effects and application thereof
CN101386823B (en) Special effect anaerobic denitrifying bacterium and waste water processing method using thereof
Wang et al. Identification and denitrification characteristics of a salt-tolerant denitrifying bacterium Pannonibacter phragmitetus F1
Meng et al. Bioconversion of wastewater by photosynthetic bacteria: Nitrogen source range, fundamental kinetics of nitrogen removal, and biomass accumulation
Lu et al. A review of ammonia-oxidizing archaea and anaerobic ammonia-oxidizing bacteria in the aquaculture pond environment in China
CN104611279B (en) A kind of red city Rhodococcus sp LH N13 and its microbial bacterial agent and purposes
Lahiri et al. Biogeochemical cycling bacteria and nutrient dynamics in waste stabilization pond system
Zhang et al. Comparision of nitrogen removal characteristic and microbial community in freshwater and marine recirculating aquaculture systems
Liu et al. Comparison of swine wastewater treatment by microalgae and heterotrophic nitrifiers: focusing on nitrogen removal mechanism revealed by microbiological correlation analysis
Das et al. Two stage treatability and biokinetic study of dairy wastewater using bacterial consortium and microalgae
CN109081447B (en) Method for removing nitrogen and phosphorus in culture wastewater by combining chlorella, acinetobacter and pseudomonas
CN103074286B (en) High-salt heterotrophic nitrification-aerobic denitrification dephosphorization salinivibrio and application of salinivibrio in wastewater treatment
CN105585133A (en) Bio-denitrification method for high-salt-content wastewater discharged from catalyst production process
CN116622556B (en) Low-temperature-resistant high-alkalinity high-efficiency denitrifying bacterium, screening method and application thereof
Qin et al. Improvement of nitrogen removal with iron scraps in floating treatment wetlands
Das et al. Biological treatment of dairy industry wastewater in a suspended growth batch reactor: performance evaluation and biodegradation kinetics
CN104263686B (en) One plant of low-temperature denitrification acinetobacter calcoaceticus and its application

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20141008

Termination date: 20170621

CF01 Termination of patent right due to non-payment of annual fee