CN104195048A - Heterotrophic-nitrification fungus capable of producing mycelium pellets, and culture method and application thereof - Google Patents
Heterotrophic-nitrification fungus capable of producing mycelium pellets, and culture method and application thereof Download PDFInfo
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- CN104195048A CN104195048A CN201410117125.2A CN201410117125A CN104195048A CN 104195048 A CN104195048 A CN 104195048A CN 201410117125 A CN201410117125 A CN 201410117125A CN 104195048 A CN104195048 A CN 104195048A
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- penicillium
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- 241000233866 Fungi Species 0.000 title claims abstract description 33
- 239000008188 pellet Substances 0.000 title claims abstract description 32
- 238000012136 culture method Methods 0.000 title abstract 2
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims abstract description 30
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 13
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000010865 sewage Substances 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims description 33
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 claims description 21
- 241000894006 Bacteria Species 0.000 claims description 19
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 12
- 229930006000 Sucrose Natural products 0.000 claims description 12
- 239000005720 sucrose Substances 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 11
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 10
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 10
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 10
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 10
- 230000001954 sterilising effect Effects 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 241000228143 Penicillium Species 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 244000005700 microbiome Species 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 6
- 230000012010 growth Effects 0.000 claims description 5
- 235000015097 nutrients Nutrition 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 3
- 238000005352 clarification Methods 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 125000001477 organic nitrogen group Chemical group 0.000 claims description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 21
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 10
- 238000004321 preservation Methods 0.000 abstract description 3
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 229910001873 dinitrogen Inorganic materials 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 abstract 1
- 238000009629 microbiological culture Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 14
- 230000001580 bacterial effect Effects 0.000 description 13
- 230000015556 catabolic process Effects 0.000 description 13
- 238000006731 degradation reaction Methods 0.000 description 13
- 239000002351 wastewater Substances 0.000 description 13
- 230000000694 effects Effects 0.000 description 10
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 229910002651 NO3 Inorganic materials 0.000 description 5
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 5
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 5
- CKUAXEQHGKSLHN-UHFFFAOYSA-N [C].[N] Chemical compound [C].[N] CKUAXEQHGKSLHN-UHFFFAOYSA-N 0.000 description 4
- 239000002054 inoculum Substances 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 238000004176 ammonification Methods 0.000 description 3
- 230000001651 autotrophic effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 125000002490 anilino group Chemical group [H]N(*)C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- -1 aromatic nitro compound Chemical class 0.000 description 2
- 238000004939 coking Methods 0.000 description 2
- 230000009514 concussion Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 230000001546 nitrifying effect Effects 0.000 description 2
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N pentanoic acid group Chemical group C(CCCC)(=O)O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000010802 sludge Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- NEAQRZUHTPSBBM-UHFFFAOYSA-N 2-hydroxy-3,3-dimethyl-7-nitro-4h-isoquinolin-1-one Chemical compound C1=C([N+]([O-])=O)C=C2C(=O)N(O)C(C)(C)CC2=C1 NEAQRZUHTPSBBM-UHFFFAOYSA-N 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- JVMRPSJZNHXORP-UHFFFAOYSA-N ON=O.ON=O.ON=O.N Chemical compound ON=O.ON=O.ON=O.N JVMRPSJZNHXORP-UHFFFAOYSA-N 0.000 description 1
- 241000228168 Penicillium sp. Species 0.000 description 1
- QTENRWWVYAAPBI-YZTFXSNBSA-N Streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@H]1[C@H](N=C(N)N)[C@@H](O)[C@H](N=C(N)N)[C@@H](O)[C@@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@H]1[C@H](N=C(N)N)[C@@H](O)[C@H](N=C(N)N)[C@@H](O)[C@@H]1O QTENRWWVYAAPBI-YZTFXSNBSA-N 0.000 description 1
- MMDJDBSEMBIJBB-UHFFFAOYSA-N [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] Chemical compound [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] MMDJDBSEMBIJBB-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
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- 238000011161 development Methods 0.000 description 1
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- 230000007613 environmental effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
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- 229910021645 metal ion Inorganic materials 0.000 description 1
- 150000002923 oximes Chemical class 0.000 description 1
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- 238000002360 preparation method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- QTENRWWVYAAPBI-YCRXJPFRSA-N streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O QTENRWWVYAAPBI-YCRXJPFRSA-N 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
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- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a heterotrophic-nitrification fungus capable of producing mycelium pellets. The heterotrophic-nitrification fungus capable of producing mycelium pellets is Penicilliumsp. L1 and is preserved in CGMCC (China General Microbiological Culture Collection Center) with the preservation number of CGMCC No. 8615. The culture method of the fungus comprises the following steps: Penicilliumsp. L1 is inoculated into a culture medium and cultured by shaking at a speed of 80-150 rpm and at a temperature of 20-38 DEG C to obtain the Penicilliumsp. L1. The fungus is used in microbial sewage containing ammonia nitrogen, and a single strain of the fungus can remove the ammonia nitrogen in the sewage and can remove nitrogen with an organic or inorganic carbon source. The strain L1 can better utilize ammonia nitrogen as the only nitrogen source under an aerobic condition to conduct nitrification, and eventually transforms nitrogen into nitrogen gas.
Description
Technical field
The present invention relates to a kind of heterotrophic nitrification, produce mycelium pellet fungi and cultural method and application, say further, is a kind of for the treatment of a strain heterotrophic nitrification fungal bacterial strain and cultural method and the application containing ammonia nitrogen microorganism in waste water.
Background technology
Nitrogenous effluent is the significant problem in Enviromental Pollution Treatment, and biological denitrificaion is to be acknowledged as at present one of most economical in denitrogenation of waste water, effective means.The bio-denitrification technology feature generally adopting is at present, denitrification process is that to make mineralized nitrogen by the coupling metabolism of nitrification and denitrification be separately nitrogen for the microorganism one Autotrophic nitrification bacterium different by two classes and heterotrophic denitrifying Bacteria.
In recent decades, obtain the multiple heterotrophic microorganism with nitrification activity from separation such as soil, crater, deep-sea, mud, lake water, included bacterium, actinomycetes and fungi etc., be called as nitrification bacteria.This is the Microbial resources that a class has significant application value, and they can utilize a lot of matrix, comprises inorganic nitrogen and organonitrogen, as ammonium, amine, acid amides, N-alkyl hydroxylamine, oxime, hydroxamic acid and aromatic nitro compound etc.
Nitrification bacteria is easy to cultivate, and propagation is very fast, and substrate utilization scope is wide, can stable existence in wastewater biological denitrificaion system.Heterotrophic nitrification process is compared with Autotrophic nitrification process, can be in lower temperature and the lower generation of oxyty (0.5 ~ 2.8mg/L), and the water quality requirement to waste water is wider, C/N can occur than more than 2.0, therefore at organic concentration and ammonia nitrogen concentration all in higher sewage disposal, heterotrophic nitrification denitrogenation with Autotrophic nitrification denitrogenation compare and there is certain advantage.Therefore adopt the denitrogenation of waste water novel process that nitrification bacteria development and Design is simple and direct, can realize quick startup and the steady running of biological denitrification process, improve nitric efficiency, reduce running cost, be expected to overcome the problem existing in conventional processes, realize the denitrogenation of waste water high-efficiency economy, for solving day by day serious nitrogenous compound, the pollution problem of environment is made contributions.
And some fungi is attended by while growth in the aqueous solution and stirs or concussion, its mycelium can be intertwined, and forms bead closely, is commonly called as mycelium pellet.Mycelium pellet is had any different in unexistent some the superior performance of other bacterial classifications.As absorption property, when with bacterium mixed culture, bacterium can be adsorbed on mycelium pellet, makes bacterium be not easy to run off, thereby strengthens some degradation property of bacterium; Biodeposition, in the time that environment leaves standstill, mycelium pellet can be deposited in bottommost, is easy to separate with water body; Decolour in addition in addition, remove the effects such as metal ion.
Summary of the invention
For above-mentioned the deficiencies in the prior art, the invention provides a strain heterotrophic nitrification, produce mycelium pellet fungi and cultural method and application.
A strain heterotrophic nitrification provided by the present invention, product mycelium pellet fungi, described in it, fungi is Penicillium
penicilliumsp
.l1, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center; Deposit number is: CGMCC No.8615.
A kind of cultural method for above-mentioned heterotrophic nitrification, product mycelium pellet fungi provided by the present invention, described in it, method is: will
penicilliumsp
.l1 is inoculated in substratum, 20-38 DEG C, 80-150rpm shaking culture; Wherein said substratum is:
Extractum carnis 3 g/L, NaCl 5g/L, peptone 10 g/L, distilled water 1000mL, regulates PH 7.2-7.4,121 DEG C of sterilizings 20 minutes; Or
Carbon source sucrose 0.5-5 g/L, nitrogenous source ammonium sulfate 0.1-2 g/L, all the other composition K
2hPO
41g/L, KCl 0.5g/L, MgSO
47H
2o 0.5g/L, FeSO
47H
2o 0.01g/L, H
2o 1000mL, tune pH is 7.0-7.4,121 DEG C of sterilizings 20 minutes; Or
Carbon source sucrose 0.5-10 g/L, aniline 0.1-2 g/L, nitrogenous source ammonium sulfate 0.1-2 g/L, all the other composition K
2hPO
41g/L, KCl 0.5g/L, MgSO
47H
2o 0.5g/L, FeSO
47H
2o 0.01g/L, H
2o 1000mL, tune pH is 7.0-7.4,121 DEG C of sterilizings 20 minutes; Or
Carbon source sucrose 0.5-10g/L, phenol 0.1-1 g/L, nitrogenous source ammonium sulfate 0.1-2 g/L, all the other composition K
2hPO
41g/L, KCl 0.5g/L, MgSO
47H
2o 0.5g/L, FeSO
47H
2o 0.01g/L, H
2o 1000mL, tune pH is 7.0-7.4,121 DEG C of sterilizings 20 minutes.
The cultural method of a kind of heterotrophic nitrification described above, product mycelium pellet fungi, the mould in described cultural method
penicilliumsp
.the bacterium colony that L1 presents on solid plate substratum is circular, and the initial stage is white, and quality is loose, after 3～5 days, is blackish green, and surface is powdery; Colony diameter is about 3.5-3.8cm, and thickness is 1-2mm.
Mould in described cultural method
penicilliumsp
.the bacterium colony that L1 presents on solid plate substratum is mycelia, mycelia prosperity, tool every, have branch; Conidiophore is shorter; Conidium is overflowed from stigma top, several not etc., You Zhouzhuan body branch.
In mould Penicillium sp. L1 liquid medium within described cultural method, produce some amount mycelium pellet, differ in size, the about 2.5mm of maximum diameter, at the bottom of can sinking to when standing bottle, nutrient solution is transparent clarification.
A kind of application for above-mentioned heterotrophic nitrification, product mycelium pellet fungi provided by the present invention, described in it, application is the ammonia nitrogen removing in sewage; And can utilize organic nitrogen source or inorganic nitrogen-sourced growth.
The application of a kind of heterotrophic nitrification described above, product mycelium pellet fungi, the best denitrogenation condition of described application is that sucrose is made carbon source, and ammonium sulfate is made nitrogenous source, and suitable C/N is 8 ~ 80, and appropriate pH is 3.0 ~ 10.0,20 ~ 40 DEG C of temperature, 120 rpms of shaking speed.
The application of a kind of heterotrophic nitrification described above, product mycelium pellet fungi, the optimum temps of applying described in it is further 30 DEG C, best initial pH value is further 7.0.
The application of a kind of heterotrophic nitrification described above, product mycelium pellet fungi, the best C/N applying described in it is 41.
Brief description of the drawings
Fig. 1 is the dull and stereotyped picture of L1 of the present invention.
Fig. 2 is L1 mycelium pellet picture of the present invention.
Fig. 3 is the spore picture under microscope of the present invention.
Embodiment
Below the specific embodiment of the present invention is further illustrated.
A strain heterotrophic nitrification provided by the present invention, product mycelium pellet fungi are to separate and obtain from the active sludge of the wastewater disposal basin of Treatment of Coking Effluent factory of Taiyuan City, are named as L1, and specifically, this bacterial strain is Penicillium
penicilliumsp
.l1, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number: CGMCC No.8615; Preservation place: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica; Postcode: 100101; The preservation time is on December 20th, 2013.
Bacterial strain in the present invention is that one has the bioactive fungi of denitrogenation, and the ammonia nitrogen that individual plant can remove in water body also can denitrogenation under organic or inorganic carbon source condition.In denitrification process, the accumulation of nitrite and nitrate do not detected.
A kind of cultural method for above-mentioned heterotrophic nitrification, product mycelium pellet fungi of the present invention, described in it, method is: will
penicilliumsp
.l1 is inoculated in substratum, 20-38 DEG C, and 80-150rpm shaking culture obtains.
Wherein said substratum is:
A, beef-protein medium: extractum carnis 3g, NaCl 5g, peptone 10g, distilled water 1000ml, 121 DEG C of sterilizings 20 minutes.
B, czapek's solution (sucrose 3g, NaNO
30.3g, K
2hPO
40.1g, KCl 0.05g, MgSO
47H
2o 0.05g, FeSO
47H
2o 0.001g, H
2o 100mL.Face the used time adds Streptomycin sulphate solution in the substratum having melted, and with anti-bacteria and actinomycetic growth, adds 1% streptomycin solution 0.3ml in every 100ml substratum)
C, heterotrophism ammonification substratum: czapek's solution (sucrose the 0.9782g, (NH of improvement
4)
2sO
40.472g, K
2hPO
40.1g, KCl 0.05g, MgSO
47H
2o 0.05g, FeSO
47H
2o 0.001g, H
2o 100ml, tune pH is 7.0-7.4.）
Above-mentioned substratum, as made solid medium, adds agar 2.0%.
Taking the active sludge of the wastewater disposal basin of Treatment of Coking Effluent factory as sample, get 10mL mud, access in sterilized above-mentioned beef extract-peptone A, in 30 DEG C, concussion enrichment culture three days in 120r/min shaking table.And then access in above-mentioned czapek's solution B, continue to cultivate three days.Get the bacterium suspension 1ml of enrichment culture in 10ml colorimetric cylinder, add sterilized water to graticule and mix, be diluted to gradient 10
-1.Again from gradient 10
-1get 1ml in 10ml colorimetric cylinder, add sterilized water to graticule and mix, be diluted to gradient 10
-2, the like.Be diluted to 10
-1~ 10
-10ten gradients, respectively get 0.2 ml dilution bacterium liquid and coat on heterotrophism ammonification Agar Plating.Coated flat board is put into biochemical cultivation case, at 30 DEG C, cultivate three days.Choose the flat board of suitable concn gradient, under aseptic condition, upper flat board every kind of bacterium colony is chosen separately respectively in the small test tube that heterotrophism ammonification liquid nutrient medium is housed, in 30 DEG C, in 120r/min shaking table, continue to cultivate.
Every 24h, from above-mentioned small test tube, get 0.5ml nutrient solution to clean ceramic whiteware dish, splash into respectively a nessler reagent, Griess-Ilosvay reagent and pentanoic reagent, compare with the blank substratum that does not connect bacterial classification, carry out denitrogenation and nitrification activity and confirm.If splash into after nessler reagent, the yellow presenting is more shallow, illustrates that in water sample, remaining ammonia-nitrogen content is fewer, and the degraded of thalline ammonia nitrogen or changing effect are better.If splash into after Griess-Ilosvay reagent, aobvious redness and even brown, illustrate in water sample and have nitrite, thalline has produced nitrite through nitrification, and the darker nitrite content of color is higher.Splash into pentanoic reagent, if aobvious blue, illustrated that nitrate exists, thalline nitrifying process has nitrate to produce, and the darker nitrate content of blueness is higher.Experimental record data are also carried out preliminary qualitative analysis, confirm the good nitrification bacteria of denitrification effect.Through enrichment, separation and purification experiment, obtain a collection of ammonia nitrogen removal effect better, nitrifying process can produce a small amount of NO
xthe bacterial strain of-N.Choose the wherein best bacterial strain of 1 strain denitrification effect, through Dalian, precious biological company limited replaces the 26S rDNA-ITS district gene sequencing carrying out, and identifies that this bacterial strain is Penicillium
penicilliumsp. name L1.
Mould
penicilliumsp
.l1 colony morphology characteristic (accompanying drawing 1): the colony characteristics that bacterial strain L1 presents on solid plate substratum: bacterium colony is for circular, and the initial stage is white, and quality is loose, after 3 to 5 days, is blackish green, and surface is powdery.Colony diameter is about 3.5-3.8cm, and thickness is 1-2mm.
Mould
penicilliumsp
.l1 morphological features (accompanying drawing 2): mycelia prosperity, tool every, have branch; Conidiophore is shorter; Conidium is overflowed from stigma top, several not etc., You Zhouzhuan body branch.
Mould
penicilliumsp
.l1 morphological features (accompanying drawing 3): in liquid medium within, produce some amount mycelium pellet, differ in size, the about 2.5mm of maximum diameter, at the bottom of can sinking to when standing bottle, nutrient solution is transparent clarification.
Bacterial strain
l1denitrification activity: bacterial strain
l1there is the ability of heterotrophism denitrogenation, and accumulate hardly nitrate and nitrite in denitrification process.
Best denitrogenation condition: sucrose is made carbon source, and ammonium sulfate is made nitrogenous source, and best C/N is 41, and optimal pH is 7.0,30 DEG C of temperature, 120 rpms of shaking speed.
Bacterial strain L1 can utilize preferably ammonia nitrogen to make only nitrogen source under aerobic condition, carries out nitrated activity, and nitrogen form is finally converted into nitrogen.
Bacterial strain of the present invention
l1be applied in sewage treatment area, in the environment of ammonia nitrogen concentration 130mg/L, through 36h growth, can be by ammonia nitrogen degradation to 5.61mg/L, ammonia nitrogen removal frank is up to 95.68%, and removing the soonest speed is 13.80mg/L/h, almost there is no the accumulation of nitrite nitrogen and nitrate nitrogen in whole process.
As ammonia nitrogen in solution 100mg/L, there is phenol 1000mg/L simultaneously, through degraded in 5 days, ammonia nitrogen in solution concentration residue 10.61mg/L, phenol concentration residue 843.84mg/L.Courseware L1 has good tolerance to high concentration phenol, and Pyrogentisinic Acid also has certain degradation capability.
As ammonia nitrogen in solution 100mg/L, there is aniline 1000mg/L simultaneously, through degraded in 5 days, ammonia nitrogen in solution concentration residue 17.94mg/L, concentration of aniline residue 352.56mg/L.Demonstrating L1 has good tolerance to high concentration aniline, and it is had to good degradation capability.
In sewage, conventionally contain some poisonous organism, L1 has good tolerance to benzene, aniline etc., has in actual applications good application prospect.
Detection method
NH
4 +-N: adopt nessler reagent light-intensity method
NO
2 --N: adopt N-(1-naphthyl)-quadrol light-intensity method
NO
3 --N: adopt ultraviolet spectrophotometry
TN:TOC-TN determinator
" water and waste water determination method " (the 4th edition) that above method is all published with reference to China Environmental Science Press.
Application Example is as follows:
(1) mould
penicilliumsp
.the Nitrification of L1 detects as follows:
Preparation simulated wastewater I, taking ammonium sulfate as only nitrogen source, starting point concentration is 130mg/L, carbon-nitrogen mass ratio is 16, is 7.0～7.4 at pH, is inoculated in 100mL fresh liquid substratum with 1% inoculum size, 30 DEG C, the interior shaking culture of 120r/min shaking table.Through 24h cultivate, by ammonia nitrogen degradation to 101.57mg/L; After 36h, final ammonia nitrogen concentration is reduced to 5.61mg/L, and degradation rate is 95.68%, and the fastest degradation rate is up to 13.8mg/L/h.
Simulated wastewater I: sucrose 4.891 g/L, (NH
4)
2sO
40.472 g/L, K
2hPO
40.5 g/L, NaCl 0.5 g/L, MgSO
47H
2o 0.5 g/L, FeSO
47H
2o 0. 01 g/L, adjusting pH is 7.0～7.4.
(2) carbon-nitrogen ratio is to mould
penicilliumsp
.the impact of L1 Nitrification
it is constant that nitrogen concentration in substratum remains 130mg/L, adjusts the carbon source quality in substratum, makes carbon-nitrogen ratio be respectively 24,32,41,48,56, and all the other conditions are constant.Be inoculated in 100mL fresh liquid substratum with 2% inoculum size, 30 DEG C, the interior shaking culture of 120r/min shaking table.After 32h, measure the residual concentration of ammonia nitrogen in solution.Institute's corresponding concentration is respectively 63.42mg/L, 30.57mg/L, 14.78 mg/L, 13.77mg/L and 12.89mg/L.Demonstrate L1 and under higher carbon-nitrogen ratio, have the better degradation effect of pair ammonia nitrogen.
(3) mould
penicilliumsp
.the research of L1 while degradation of ammonia nitrogen and aniline
In simulated wastewater I, add in addition aniline, make concentration of aniline be respectively 200mg/L, 400mg/L, 600mg/L, 800mg/L and 1000mg/L.With 2% inoculum size inoculation, 30 DEG C, the interior shaking culture of 120r/min shaking table.After 5d, measure ammonia nitrogen in solution and concentration of aniline.Ammonia nitrogen concentration is respectively 3.18mg/L, 12.43mg/L, 13.74mg/L, 16.55mg/L, 17.94mg/L; Concentration of aniline is respectively 149.67mg/L, 328.78mg/L, 545.18mg/L, 490.49mg/L, 352.56mg/L.Demonstrate L1 degradation of ammonia nitrogen and the good ability of aniline simultaneously.
(4) mould
penicilliumsp
.the research of L1 while degradation of ammonia nitrogen and phenol
In simulated wastewater I, add in addition phenol, make phenol concentration be respectively 200mg/L, 600mg/L and 1000mg/L.With 2% inoculum size inoculation, 30 DEG C, the interior shaking culture of 120r/min shaking table.After 2d, ammonia nitrogen in solution concentration remains respectively 9.85mg/L, 11.37mg/L, 10.61mg/L; After 5d, Phenol in Aqueous Solution concentration remains respectively 95.27mg/L, 391.59 mg/L, 843.84mg/L.Even if illustrate that L1 still keeps good ammonia nitrogen degradation ability in the situation that high concentration phenol exists, and Pyrogentisinic Acid also has certain degradation capability.
sequence table
<110> Institutes Of Technology Of Taiyuan
<120> mono-strain heterotrophic nitrification fungi and application method and purposes
<130> patent application
<160>1
<210>1
<211>528
<212>DNA
<213> Penicillium (
penicillium.)
<400>
gcgggcccct cgtggcccaa cctccccacc cttgtctcta tacacctgtt gctttggcgg 60
gcccaccggg gccacctggt cgccggggga cgttcgtccc cgggcccgcg cccgccgaag 120
cgctctgtga accctgatga agatgggctg tctgagtact atgaaaattg tcaaaacttt 180
caacaatgga tctcttggtt ccggcatcga tgaagaacgc agcgaaatgc gataagtaat 240
gtgaattgca gaattccgtg aatcatcgaa tctttgaacg cacattgcgc cccctggcat 300
tccggggggc atgcctgtcc gagcgtcatt tctgccctca agcacggctt gtgtgttggg 360
tgcggtcccc ccgggggcct gcccgaaagg cagcggcgac gtccgtctgg tcctcgagcg 420
tatggggctt tgtcactcgc tcgggaagga ctggcggggg ttggtcacca ccacaaaatt 480
ttaccacggt tgacctcgga tcaggtagga gttacccgct gaacttaa 528
Claims (9)
1. a strain heterotrophic nitrification, product mycelium pellet fungi, described in it, fungi is Penicillium
penicilliumsp
.l1, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center; Deposit number is: CGMCC No.8615.
2. for a cultural method for the heterotrophic nitrification described in claim 1, product mycelium pellet fungi, described in it, method is: will
penicilliumsp
.l1 is inoculated in substratum, 20-38 DEG C, 80-150rpm shaking culture; Wherein said substratum is:
Extractum carnis 3 g/L, NaCl 5g/L, peptone 10 g/L, distilled water 1000mL, regulates PH 7.2-7.4,121 DEG C of sterilizings 20 minutes; Or
Carbon source sucrose 0.5-5 g/L, nitrogenous source ammonium sulfate 0.1-2 g/L, all the other composition K
2hPO
41g/L, KCl 0.5g/L, MgSO
47H
2o 0.5g/L, FeSO
47H
2o 0.01g/L, H
2o 1000mL, tune pH is 7.0-7.4,121 DEG C of sterilizings 20 minutes; Or
Carbon source sucrose 0.5-10 g/L, aniline 0.1-2 g/L, nitrogenous source ammonium sulfate 0.1-2 g/L, all the other composition K
2hPO
41g/L, KCl 0.5g/L, MgSO
47H
2o 0.5g/L, FeSO
47H
2o 0.01g/L, H
2o 1000mL, tune pH is 7.0-7.4,121 DEG C of sterilizings 20 minutes; Or
Carbon source sucrose 0.5-10g/L, phenol 0.1-1 g/L, nitrogenous source ammonium sulfate 0.1-2 g/L, all the other composition K
2hPO
41g/L, KCl 0.5g/L, MgSO
47H
2o 0.5g/L, FeSO
47H
2o 0.01g/L, H
2o 1000mL, tune pH is 7.0-7.4,121 DEG C of sterilizings 20 minutes.
3. the cultural method of a kind of heterotrophic nitrification as claimed in claim 2, product mycelium pellet fungi, mould described in it
penicilliumsp
.the bacterium colony that L1 presents on solid plate substratum is circular, and the initial stage is white, and quality is loose, after 3～5 days, is blackish green, and surface is powdery; Colony diameter is about 3.5-3.8cm, and thickness is 1-2mm.
4. the cultural method of a kind of heterotrophic nitrification as claimed in claim 2, product mycelium pellet fungi, mould described in it
penicilliumsp
.the bacterium colony that L1 presents on solid plate substratum is mycelia, mycelia prosperity, tool every, have branch; Conidiophore is shorter; Conidium is overflowed from stigma top, several not etc., You Zhouzhuan body branch.
5. the cultural method of a kind of heterotrophic nitrification as claimed in claim 2, product mycelium pellet fungi, mould Penicillium sp described in it
.in L1 liquid medium within, produce some amount mycelium pellet, differ in size, the about 2.5mm of maximum diameter, at the bottom of can sinking to when standing bottle, nutrient solution is transparent clarification.
6. an application for a strain heterotrophic nitrification as claimed in claim 1, product mycelium pellet fungi, described in it, application is the ammonia nitrogen removing in sewage; And can utilize organic nitrogen source or inorganic nitrogen-sourced growth.
7. the application of a strain heterotrophic nitrification as claimed in claim 6, product mycelium pellet fungi, the best denitrogenation condition of applying described in it is that sucrose is made carbon source, ammonium sulfate is made nitrogenous source, suitable C/N is 8 ~ 80, appropriate pH is 3.0 ~ 10.0,20 ~ 40 DEG C of temperature, 120 rpms of shaking speed.
8. the application of a strain heterotrophic nitrification as claimed in claim 7, product mycelium pellet fungi, the optimum temps of applying described in it is further 30 DEG C, best initial pH value is further 7.0.
9. the application of a strain heterotrophic nitrification as claimed in claim 7, product mycelium pellet fungi, the best C/N applying described in it is 41.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117887593A (en) * | 2024-03-13 | 2024-04-16 | 西安建筑科技大学 | Mixed nutrition type denitrifying bacterium Penicillium sp.N8 and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102476865A (en) * | 2010-11-26 | 2012-05-30 | 中国辐射防护研究院 | Wastewater denitrogenation method by use of penicillium strain |
| CN103484378A (en) * | 2013-08-09 | 2014-01-01 | 太原理工大学 | Heterotrophic nitrification aerobic denitrification fungus as well as cultural method and application thereof |
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2014
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102476865A (en) * | 2010-11-26 | 2012-05-30 | 中国辐射防护研究院 | Wastewater denitrogenation method by use of penicillium strain |
| CN103484378A (en) * | 2013-08-09 | 2014-01-01 | 太原理工大学 | Heterotrophic nitrification aerobic denitrification fungus as well as cultural method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 吕永康等: "一株异养硝化细菌的筛选及其硝化特性研究", 《PROCEEDING OF 2010 FIRST INTERNATIONAL CONFERENCE ON CELLULAR,MOLECULAR BIOLOGY,BIOPHYSICS AND BIOENGINEERING》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117887593A (en) * | 2024-03-13 | 2024-04-16 | 西安建筑科技大学 | Mixed nutrition type denitrifying bacterium Penicillium sp.N8 and application thereof |
| CN117887593B (en) * | 2024-03-13 | 2024-05-31 | 西安建筑科技大学 | Mixed nutrition type denitrifying bacterium Penicillium sp.N8 and application thereof |
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