CN103468586A - Penicillium strain and application in treatment of sewage and odor pollution thereof - Google Patents

Penicillium strain and application in treatment of sewage and odor pollution thereof Download PDF

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CN103468586A
CN103468586A CN2013103844163A CN201310384416A CN103468586A CN 103468586 A CN103468586 A CN 103468586A CN 2013103844163 A CN2013103844163 A CN 2013103844163A CN 201310384416 A CN201310384416 A CN 201310384416A CN 103468586 A CN103468586 A CN 103468586A
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sewage
ammonia
degradation
bacterial strain
state nitrogen
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靳永胜
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Beijing Huanyang Environmental Protection Science & Technology Development Co Ltd
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Abstract

The invention belongs to the field of microbiology, in particular to a penicillium strain which can efficiently degrade wastewater and ammonia nitrogen in odor, and the application in treatment of sewage and odor pollution thereof. Penicillium, also Penicillium purpurogenum by classification, has been preserved in the China General Microbiological Culture Collection Center (CGMCC), CGMCC No. 8036. The bacterial strain has the advantages of efficiently degrading wastewater and ammonia nitrogen in odor, and efficiently treating sewage and odor pollution.

Description

A kind of penicillium bacterial strain and the application in administering sewage and odor pollution thereof
Technical field
The invention belongs to microorganism field, be specifically related to penicillium bacterial strain and the application in administering sewage and odor pollution thereof of ammonia-state nitrogen in a kind of efficient degradation sewage and foul smell.
Background technology
Along with the continuous acceleration of socioeconomic develop rapidly and urbanization process, the indoor and outdoor surroundings pollution problem becomes increasingly conspicuous, the primary factor that especially day by day serious water pollution and topsoil become puzzlement and threaten human health and daily life.No matter be water pollution or topsoil, smell that it is caused by pollutent pollute be stench not only the stimulating human olfactory organ cause the unhappy also entail dangers to damage coenocorrelation and human health of emotional injury that waits of people.
Ammonia-state nitrogen is to be decomposed in soil and contained the nitrogenous source that animals and plants remains and excremental protein and uric acid, urea etc. draw by microorganism.Ammonia-state nitrogen is the Main Morphology of nitrogen in aqueous environment, is a kind of important pollutent of body eutrophication and environmental pollution, is also the major cause that causes producing in sewage and atmosphere odor pollution.
External some country has just started the research to the improvement aspect of odor pollution earlier, stench is carried out to special legislation, using odor pollution as a kind of public hazards.Deodour method is mainly Physical, chemical method and biological process at present.But wherein Physical, chemical method often exist the problem that apparatus expensive, working cost are high, secondary pollution is arranged, thereby the foreign scholar just is devoted to the exploitation of microbial deodorization method from the 1950's.
Biological deodorization method refers to and utilizes the microbiological deterioration repugnant substance, is mainly the ammonia-state nitrogen material in sewage and foul smell, reaches the method for removing stink.Utilize biological process dispose of sewage and atmosphere in foul gas report the earliest be in nineteen sixty, early 1970s, the scholar of European Section started the research of this respect; The eighties, Holland and Germany utilized the microbiological treatment foul gas to obtain good effect, caused subsequently the broad research of the U.S., Japan and other Western European countries.Current many developed countries are as the commercialization of exploitation of the biological deodorization technology and equipments such as Japan, the U.S., Germany.China just starts to carry out theory and the paractical research of this respect the initial stage nineties.But nearly ten years, the many researchists of China made outstanding contribution to the development of its Science and Technology, all obtaining gratifying achievement aspect theory and technology, technique and equipment.From now on, along with the development of biological deodorization technology and various correlative studys thereof, the microbial deodorization method will be more and more universal.
Due to general microorganism can only utilize and degradation water in deliquescent material, at first the repugnant substance therefore be degraded will be dissolved in the water, then is transferred in microbe, self Metabolic activity by microorganism is degraded.Therefore, microorganism becomes the key of microbial deodorization method for the degradation capability of ammonia-state nitrogen class material.Therefore, to promote the fast development of microbial deodorization technology, and advance the improvement of sewage and odor pollution in the urgent need to the microorganism strains of developing suitable extensive use and stable performance, degradation capability are strong.
Summary of the invention
For this reason, technical problem to be solved by this invention be to provide a kind of can efficient degradation sewage and foul smell in the penicillium bacterial strain of ammonia-state nitrogen, and provide the application of described penicillium bacterial strain in administering sewage and odor pollution.
The problems referred to above are achieved through the following technical solutions:
The inventor is through screening, select a strain of penicillium bacterial strain, its Classification And Nomenclature is penicillium purpurogenum Penicillium purpurogenum, be preserved in Chinese microorganism strain management committee common micro-organisms center (being called for short CGMCC), depositary institution address: China. No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, deposit number is CGMCC8036, preservation date is on August 16th, 2013, the bacterial strain that this bacterial strain is ammonia-state nitrogen in degradable sewage and foul smell.
This bacterial strain has following character: the bacterium colony of described penicillium bacterial strain is on the LB flat board, and the fine hair sample bacterium colony that is creamy white in early days, the cyan spore occurs on later stage bacterium center falls, and mycelia becomes light red.
The present invention also provides a kind of biological bacteria degradation agents, and it comprises described penicillium bacterial strain.
The invention also discloses the application in administering sewage and odor pollution field of described penicillium bacterial strain and described biological bacteria degradation agents.
The invention also discloses the method for ammonia-state nitrogen in a kind of degradation of sewage and foul smell, comprise and use described penicillium bacterial strain and described biological bacteria degradation agents and the contacted step of described ammonia-state nitrogen.
The method of ammonia-state nitrogen in described degradation of sewage and foul smell, also comprise the step that described penicillium bacterial strain is cultivated in the fermention medium of suitable its growth.
Concrete, described fermention medium contains following content component: glucose 3-8g/L, (NH) 2sO 40.4-0.8g/L, NaCl0.5-1.5g/L, FeSO 47H 2o 0.02-0.08g/L, K 2hPO 40.3-0.8g/L, MgSO 47H 2o 0.2-0.3g/L, all the other are water; Perhaps, brown sugar 8-12g/L, ammonium sulfate 1-3g/L, all the other are water; Perhaps, tryptone 8-12g/L, yeast extract 3-8g/L, sodium-chlor 8-12g/L, all the other are water.
Further, described fermention medium contains following content component: glucose 5g/L, (NH) 2sO 40.6g/L, NaCl 1.0g/L, FeSO 47H 2o 0.05g/L, K 2hPO 40.5g/L, MgSO 47H 2o0.25g/L, all the other are water; Perhaps, brown sugar 10g/L, ammonium sulfate 2g/L, all the other are water; Perhaps, tryptone 10g/L, yeast extract 5g/L, sodium-chlor 10g/L, all the other are water.
Described culture condition is: control temperature 25-30 ℃, pH6-8, rotating speed 150-200rpm, fermentation time 36-60 hour.
[0018] the present invention obtains a strain through screening and the ammonia-state nitrogen pollutant matter that causes sewage and odor pollution is had to the penicillium bacterial strain Penicilliumpurpurogenum BJHY-501 of extremely strong degradation capability, and its ammonia-state nitrogen mass degradation ability reaches and approaches 98%; And described bacterial strain is better to the Governance Ability of contaminated sewage and foul smell, can effectively solve the environmental improvement of sewage stench and foul smell.
The accompanying drawing explanation
Fig. 1 is nessler reagent colorimetric method for determining NH in embodiment 3 in the present invention 4 +the typical curve that-N draws.
Embodiment
The concrete composition of the substratum that described in following each embodiment, screening and culturing step relate to comprises:
(1) de-NH 3bacterial liquid screening culture medium component: glucose 5g, (NH) 2sO 40.6g, NaCl 1.0g, FeSO 47H 2o 0.05g, K 2hPO 40.5g, MgSO 47H 2o 0.25g, deionized water is settled to 1000ml, 121 ℃ of sterilizing 30min;
(2) de-NH 3bacterial solids screening culture medium component: glucose 5g, (NH) 2sO 40.6g, NaCl 1.0g, FeSO 47H 2o 0.05g, K 2hPO 40.5g, MgSO 47H 2o 0.25g, agar powder 15g, deionized water is settled to 1000ml, 121 ℃ of sterilizing 30min;
(3) LB liquid nutrient medium: tryptone 10g, yeast extract 5g, sodium-chlor 10g, deionized water is settled to 1000ml, 121 ℃ of sterilizing 30min;
(4) LB solid medium: tryptone 10g, yeast extract 5g, sodium-chlor 10g, agar powder 15g, deionized water is settled to 1000ml, 121 ℃ of sterilizing 30min;
(5) de-NH 3bacterium enlarged culturing nutrient media components: brown sugar 10g, ammonium sulfate 2g, deionized water is settled to 1000ml, 121 ℃ of sterilizing 30min.
Embodiment 1 bacterial screening
Get respectively 10ml Beijing Nangong garbage compost factory percolate 20 lot samples, be inoculated into respectively in the shaking flask that 100ml LB liquid nutrient medium is housed, cultivate 2 days on constant temperature oscillator, temperature is 30 ℃, and rotating speed is 180r/min.Observe the interior liquid of bottle muddy to occurring, present oyster white, now bacteria growing is the most vigorous.
For whether containing the bacterial classification of degradation of ammonia nitrogen in the preliminary evaluation sample, each screening sample is carried out to following mensuration:
The described de-NH of 100ml packs in the Erlenmeyer flask of 250ml 3the bacterial liquid screening culture medium, do multiple batches of Duplicate Samples, and do blank sample contrast;
Respectively the supernatant liquor 200ul of the sample to be screened of aforementioned preliminary cultivation be inoculated in prepared containing the de-NH of 100ml 3in the screening culture medium of bacterial liquid, at rotating speed, be 180r/min, cultivate 5 days on the vibration shaking table that temperature is 30 ℃, result checks to solution and becomes muddy at any time, uses nessler reagent colorimetric method for determining NH every day 4 +the content of-N, if find NH in sample 4 +-N content reduces, and shows to have the bacterial classification of degradation of ammonia nitrogen to exist.
Get the supernatant nutrient solution that preliminary judgement contains in the sample that degradable ammonia nitrogen bacterial classification exists and be inoculated in described de-NH 3on bacterial solids screening culture medium flat board, treat on flat board to grow bacterium colony, choose the different bacterium colonies purifying of repeatedly ruling on flat board, its step is as follows:
(1) line separates: nutrient solution is inoculated in to de-NH 3on bacterial solids screening culture medium flat board, treat on flat board to grow single bacterium colony, the different bacterium colonies of picking, on flat board, line separates repeatedly, and approximately after 4 weeks, obtain 13 strain Y1, Y2, Y3, Y4, Y5, Y6, Y7, Y8, Y9, Y10, Y11, Y12, Y13 final the separation;
(2) line purifying, the experimental strain after screening obtains different single bacterium colonies after plate loop method repeatedly, until under microscope, check, without miscellaneous bacteria, is the bacterial classification of purifying;
(3) culture presevation, be inoculated into described de-NH to bacterial classification after purifying 3in the bacterial liquid screening culture medium, each bacterial strain of test is under 30 ℃, 180rpm condition, cultivates after 48h NH 4 +-N utilizes situation, with nessler reagent colorimetric method for determining NH 4 +the content of-N is index, filters out NH 4 +-N transforms good bacterial strain;
(4) bacterial strain that the selection transformation efficiency is higher, as the purpose bacterial strain, is seeded on described LB slant medium, cultivates 48h for 30 ℃ in constant incubator, puts into 4 ℃ of refrigerators and preserves, and carries out renewed vaccination every two months and cultivates preservation.
The bacterial strain that resulting vigor is higher carries out the genetic stability investigation.By sieve to obtain bacterial strain continuous 10 generations of switching, in per generation, all accessed described de-NH 3carry out cultivation and fermentation in the bacterial liquid screening culture medium, the bacterial strain that sieves has shown stable leavening property.The bacterial strain called after BJHY-501 that wherein degradation capability is higher, stability is best.
Bacterial strain BJHY-501 is carried out to DNA sequencing, and sequencing result and corresponding database are compared, pass through sequencing and carry out sequence alignment on http://archive-dtd.ncbi.nlm.nih.gov/ website, the sequence homology of finding the gene order of this bacterial strain and registered Penicillium notatum (Penicilliumpurpurogenum) is the highest, reaches 100%.Thereby determine that this bacterial strain is Penicillium notatum.
The bacterial strain called after penicillium purpurogenum Penicillium purpurogenum that screening is obtained, and be preserved in Chinese microorganism strain management committee common micro-organisms center (be called for short CGMCC), deposit number is CGMCC8036, the bacterial strain that this bacterial strain is ammonia-state nitrogen in degradable sewage and foul smell.
The enlarged culturing of embodiment 2 bacterial strains
The fermentation culture method of this bacterial classification is as follows:
(1) at first by the described de-NH of bacterial strain BJHY-501 streak inoculation of freezing preservation 3cultivate 2-3 days in 28-30 ℃ of incubator on the bacterial solids screening culture medium, grow single bacterium colony;
(2) adopt de-NH 3the bacterial liquid screening culture medium is dug single colony inoculation from the solid plate substratum and prepare seed liquor in Erlenmeyer flask, and under 28-30 ℃, rotating speed is 180r/min, and 48-36 hour is cultivated in concussion;
(3) according to 10-20%(v/v) inoculum size described seed liquor is seeded to de-NH 3enlarged culturing in bacterium enlarged culturing substratum, 28-30 ℃, rotating speed 150-200rpm, aeration is cultivated and can be used in 2-3 days.After fermentation ends, the dry weight of test thalline is 15.26g/L.
Embodiment 3 strains for degrading ammonia-state nitrogen abilities are measured
With nessler reagent colorimetric method for determining NH 4 +the method of-N
The configuration of reagent
NH 4 +-N standard reserving solution: take 3.819g NH 4cl is dissolved in the water, and is settled to 1000ml, is equivalent to the NH that contains 1000ug in every ml water 4 +-N.
The measuring method of typical curve: according to following table configuration solution, after configuring, each pipe adds 0.2ml sodium tartrate potassium solution, shakes up, then adds the nessler reagent of 0.2ml, and room temperature is placed 10min, and then measures light absorption value A under the 420nm wavelength 420, think A 420x-coordinate, ammonia-nitrogen content is that ordinate zou is set up regression curve, is typical curve.
The drafting of typical curve
Figure BDA00003741971700071
Take transverse axis as absorbancy, and the longitudinal axis is ammonia-nitrogen content drawing standard curve, as shown in Figure 1.
NH 4 +-N is used liquid: draw 5mlNH 4 +-N standard reserving solution is settled in 500ml water, is equivalent to the NH that contains 10ug in every ml water 4 +-N.
Get the fermented liquid of enlarged culturing in step (3), according to 10%(v/v) inoculum size access described NH 4 +-N is used the degraded of carrying out ammonia nitrogen in liquid to measure, and every 24h of take more than 6 days, the results are shown in following table as the interval METHOD FOR CONTINUOUS DETERMINATION:
My god 1 2 3 4 5 6
The ammonia nitrogen degradation rate 2.63% 30.98% 61.03% 90.90% 97.70% 97.77%
Experiment shows, the penicillium bacterial strain BJHY-501 that described screening obtains has ammonia-state nitrogen degradation rate preferably, and its degradation rate of about 6 days is up to nearly 98%, and effect is remarkable.
Following experimental example is the technique effect of provable scheme of the present invention all.
The improvement experimental example of described bacterial strain to sewage and odor pollution
Following each experimental example be take in embodiment 2 and is cultivated the fermented liquid that obtains and carry out compliance test result as example, according to 10%(v/v) inoculum size inoculated.
Experimental example 1:
Get the Nangong composting plant percolate 50L of Beijing as test sample book, and measure its corresponding contamination index, specifically comprise CODcr:35000mg/L, BOD5:22000mg/L, suspended substance: 2600mg/L, NH3-N:1800mg/L, initial percolate distributes unpleasant foul smell.
Access described the confession and, in examination fermented liquid test sample book, in the plastic tank of a 200L, utilize a miniature aerator continuous aeration (320L/ hour), normal temperature is processed 4 days.
After question response finishes, described sewage sample through degradation treatment is carried out to index test, specifically comprise CODcr:12750mg/L, BOD5:8560mg/L, suspended substance 1040mg/L, NH3-N:162mg/L, degradation rate is respectively 63.57%, 61%, 60%, 91%, and the foul smell of percolate fades away.
Experimental example 2:
Circulation bacterium liquid using the bacterium liquid of described bacterial strain as the bio-filtration deodorization tower, the volcanics that the deodorizing tower filler is the 30-50mm particle diameter or plastics Pall ring, depth of packing is at 60-120cm, spray flux 1.5-3L/ ㎡. minute, bacterium liquid circulated sprinkling can hang up mycoderm in one week on filler, and the bio-trickling filter after biofilm reaches more than 99% the clearance of ammonia in garbage compost factory fermentation plant foul smell.
Experimental example 3:
Circulation cleaning bacterium liquid using the bacterium liquid of described bacterial strain as biological washing tower, the washing tower filler is the plastics Pall ring, and depth of packing is 80-120cm, and washing amount is 30-50L/ ㎡. minute, the clearance of ammonia in garbage compost factory fermentation plant foul smell is reached more than 99%.
The explanation of above-mentioned test result, bacterial strain of the present invention is the ammonia-state nitrogen material in degradation of sewage effectively, the sewage stench that effectively alleviation causes because of ammonia-state nitrogen and the pollution of foul smell, not only regulation effect is better, and practice of harnessing is shorter.
Obviously, above-described embodiment is only for example clearly is described, and is not the restriction to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without also giving all embodiments.And the apparent variation of being extended out thus or change are still among the protection domain in the invention.

Claims (10)

1. a penicillium bacterial strain, its Classification And Nomenclature is Penicilliumpurpurogenum BJHY-501, has been preserved in Chinese microorganism strain management committee common micro-organisms center, and deposit number is CGMCC 8036, and preservation date is on August 16th, 2013.
2. a biological bacteria degradation agents, is characterized in that, comprises penicillium bacterial strain claimed in claim 1.
3. the application of penicillium bacterial strain claimed in claim 1 in administering sewage and odor pollution field.
4. the application of biological bacteria degradation agents claimed in claim 2 in administering sewage and odor pollution field.
5. the method for ammonia-state nitrogen in a degradation of sewage and foul smell, is characterized in that, comprises that right to use requires 1 described penicillium bacterial strain and the contacted step of described ammonia-state nitrogen.
6. the method for ammonia-state nitrogen in degradation of sewage according to claim 5 and foul smell, is characterized in that, also comprises the step that described penicillium bacterial strain is cultivated in the fermention medium of suitable its growth.
7. the method for ammonia-state nitrogen in degradation of sewage according to claim 6 and foul smell, is characterized in that, described fermention medium contains following content component: glucose 3-8g/L, (NH) 2sO 40.4-0.8g/L, NaCl0.5-1.5g/L, FeSO 47H 2o 0.02-0.08g/L, K 2hPO 40.3-0.8g/L, MgSO 47H 2o 0.2-0.3g/L, its be water; Perhaps, brown sugar 8-12g/L, ammonium sulfate 1-3g/L, all the other are water; Perhaps, tryptone 8-12g/L, yeast extract 3-8g/L, sodium-chlor 8-12g/L, all the other are water.
8. the method for ammonia-state nitrogen in degradation of sewage according to claim 7 and foul smell, is characterized in that, described fermention medium contains following content component: glucose 5g/L, (NH) 2sO 40.6g/L, NaCl 1.0g/L, FeSO 47H 2o 0.05g/L, K 2hPO 40.5g/L, MgSO 47H 2o 0.25g/L, its be water; Perhaps, brown sugar 10g/L, ammonium sulfate 2g/L, all the other are water; Perhaps, tryptone 10g/L, yeast extract 5g/L, sodium-chlor 10g/L, all the other are water.
9. according to the method for ammonia-state nitrogen in the described degradation of sewage of claim 7 or 8 and foul smell, it is characterized in that, described culture condition is: control temperature 25-30 ℃, pH6-8, rotating speed 150-200rpm, fermentation time 36-60 hour.
10. the method for ammonia-state nitrogen in a degradation of sewage and foul smell, is characterized in that, comprises that right to use requires 2 described biological bacteria degradation agentss and the contacted step of described ammonia-state nitrogen.
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CN109317514A (en) * 2018-12-05 2019-02-12 湖南省农业生物技术研究所 Penicillium bacterial strain CN35 is in the application administered in heavy metal pollution
CN116925934A (en) * 2022-03-31 2023-10-24 南京农业大学 Bacterial strain for degrading odor substance n-butyric acid and application thereof

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CN109317514A (en) * 2018-12-05 2019-02-12 湖南省农业生物技术研究所 Penicillium bacterial strain CN35 is in the application administered in heavy metal pollution
CN116925934A (en) * 2022-03-31 2023-10-24 南京农业大学 Bacterial strain for degrading odor substance n-butyric acid and application thereof
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Application publication date: 20131225