CN104560809B - A kind of Staphylococcus pasteuri KY 06 and application - Google Patents
A kind of Staphylococcus pasteuri KY 06 and application Download PDFInfo
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- CN104560809B CN104560809B CN201410836858.1A CN201410836858A CN104560809B CN 104560809 B CN104560809 B CN 104560809B CN 201410836858 A CN201410836858 A CN 201410836858A CN 104560809 B CN104560809 B CN 104560809B
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Abstract
The invention provides a kind of Staphylococcus pasteuri KY 06, preserving number is CGMCC No.10173, degradation capability with stronger sludge organic matter, the degraded of complicated organic matter in anaerobic digestion reaction system can be significantly improved under anaerobic, the degraded of organic matter and gas production in anaerobic sludge digestion reaction can be significantly improved, and help the stability for maintaining anaerobic digestion reaction system, have cost low compared with chemical method, the advantages of non-secondary pollution, culture is easy to use, it can be industrial wastewater biologic treating technique, in particular positive contribution is made in the further genralrlization of anaerobic digestion techniques and popularization, it is with a wide range of applications in sewage treatment area.
Description
Technical field
The present invention relates to biological technical field, especially a kind of Staphylococcus pasteuri KY-06 and application.
Background technology
Sludge is the accessory substance in sewage disposal process, with industry and the increasingly raising of living standards of the people, sewage
Yield is continuously increased, and causes sludge to increase with every year about 10% speed.Because moisture percentage in sewage sludge is high, containing a large amount of organic matters,
Perishable, stench and containing parasitic ovum, pathogenic microorganism and heavy metal etc., if do not dealt carefully with, is arbitrarily discharged,
Secondary pollution will be caused.Therefore minimizing, innoxious and recycling treatment need to be carried out to sludge.And anaerobic digestion is using simultaneous
Oxygen bacterium and anaerobic bacteria decompose organic matter in sludge under anaerobic environment, are the volume reduction process for stabilizing commonly used in sludge treatment technique,
And have the advantages that energy consumption is low, stability is good, produce biogas, therefore worldwide receive significant attention.But anaerobism
Digestion also faces many difficulties, and such as hydrolysis rate is slow, and organism utilization rate is low, and the anaerobic digestion residence time is long, factor of created gase
Low problem.
Generally believe that anaerobic digestion is divided into hydrolysis, acidifying and aerogenesis three phases, due to containing a large amount of difficult degradations in sludge
Organic matter such as cellulose etc., the degraded of organic matter is the rate-limiting step of anaerobic sludge digestion.In order to improve anaerobic digestion speed
And gas production, different researchers propose respectively optimization solve scheme.(1) to anaerobic System addition metal ion, such as K,
Mg, Fe, Co, Ni etc., metal ion can accelerate the degraded of organic matter to a certain extent, improve gas production；(2) disappear to anaerobism
Change system adds some inorganic matters, such as PAM, can excite the activity of methanogen, methane phase tolerance is improved, while anaerobic processes
Middle PAM can be also utilized as nitrogen source；(3) biology enzyme is added into sludge, such as addition promotes the enzyme of cell membrane hydrolysis can be with
Promote digestion process and increase gas production, enzyme agent is added in sludge can improve the bioactivity of anaerobe, and raising is detested
The dewatering of sludge organism body during oxygen.
As described above, adding the methods such as metal ion, inorganic matter, biology enzyme into anaerobic digester system, on the one hand increase
Specific resistance to filtration cost, while unnecessary interference can be produced to operating anaerobic digester system.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of Staphylococcus pasteuri KY-06, with stronger fiber
Plain degradation capability.
Another technical problem to be solved by this invention is the application for providing above-mentioned Staphylococcus pasteuri KY-06.
In order to solve the above technical problems, the technical scheme is that：
A kind of Staphylococcus pasteuri (Staphylococcus pasteuri) KY-06, its preserving number is CGMCC
No.10173 (depositary institutions:China Committee for Culture Collection of Microorganisms's common micro-organisms center, address：Beijing is exposed to the sun
The institute 3 of area North Star West Road 1, preservation date：December 15, deposit number in 2014：CGMCC No.10173).
It is preferred that, above-mentioned Staphylococcus pasteuri KY-06, thalline is in spherical, and diameter is about 0.5~1.5 μm, gram sun
Property, no gemma is identified by 16S rDNA and Biolog microorganism classifications identification systems, is defined as Staphylococcus pasteuri
(Staphylococcus pasteuri), is named as Staphylococcus pasteuri KY-06, and its systematic evolution tree is as shown in figure 1,16S
RDNA the sequencing results is in sequence tables<400>Sequence shown in 1, i.e.,：
5’-AAAGTGGAATTCCATGTGTAGCGGTGAAATGCGCAGAGATATGGAGGAACACCAGTGGCGAAGGCG
ACTTTCTGGTCTGTAACTGACGCTGATGTGCGAAAGCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGC
CGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGG
GGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCG
AAGCAACGCGAAGAACCTTACCAAATCTTGACATCCTTTGACCGCTC-3’。
It is preferred that, above-mentioned Staphylococcus pasteuri KY-06, the V3 that the nucleotide sequence belongs to bacterial 16 S rDNA sequences can
Become area.
It is preferred that, above-mentioned Staphylococcus pasteuri KY-06 is isolated from the ruminant stomach of ox, bacterium is shown as in screening flat board
It is in faint yellow to fall, and neat in edge, surface is smooth, convex protuberance.
It is preferred that, above-mentioned Staphylococcus pasteuri KY-06, the degradation capability with stronger sludge organic matter.
Above-mentioned Staphylococcus pasteuri KY-06 produces the synergy purposes of natural pond reaction to anaerobic digestion.
It is preferred that, the bacterial strain Staphylococcus pasteuri KY-06 is seeded to by above-mentioned Staphylococcus pasteuri KY-06 application
In enriched medium, 30 DEG C of shaken cultivations are to increased logarithmic phase, bacterium dense about 106~1010Individual/ml, centrifuging and taking bacterium solution supernatant, is pressed
Catabolism ability of the supernatant to organic matters such as celluloses is determined according to DNS reduced sugar colorimetric National Standard Method GB/T 23881-2009,
And be inoculated in 0.1~0.5% inoculum concentration in anaerobic digestion reaction system, realize the synergy that the bacterial strain is reacted anaerobic digestion
Effect.
The beneficial effects of the invention are as follows：
, can be under anaerobic the invention provides Staphylococcus pasteuri KY-06 (preserving number is CGMCC No.10173)
The degraded of complicated organic matter in anaerobic digestion reaction system is significantly improved, can be significantly improved organic in anaerobic sludge digestion reaction
The degraded of matter and gas production, and help maintain anaerobic digestion reaction system stability, with chemical method compared with cost it is low,
The advantages of non-secondary pollution, cultivate easy to use, can be industrial wastewater biologic treating technique, in particular anaerobic digestion techniques are entered
One step, which is popularized, makes positive contribution, is with a wide range of applications in sewage treatment area.
Brief description of the drawings
Fig. 1 is Staphylococcus pasteuri KY-06 of the present invention systematic evolution tree.
Fig. 2 be Staphylococcus pasteuri KY-06 of the present invention to municipal sludge aerogenesis synergistic results figure, wherein, above it is bent
Line is municipal sludge+bacterium, and lower surface curve is municipal sludge.
Preservation information
Classification noun：Staphylococcus pasteuri (Staphylococcus pasteuri)
Depositary institution's title：China Committee for Culture Collection of Microorganisms's common micro-organisms center
Depositary institution address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date：On December 15th, 2014
Preserving number：CGMCC No.10173
Embodiment
Technical scheme of the present invention is further described with reference to specific embodiment.
Different culture mediums and solution formula are as follows in following embodiments：
1. screening and culturing medium：15~30g of cellulose, potassium dihydrogen phosphate 2g, 1~2g of ammonium sulfate, 0.2~1g of magnesium sulfate, chlorine
Change 0.2~1g of calcium, 3~6mg of ferrous sulfate, 1~2mg of manganese sulfate, 1.0~1.7mg of zinc chloride, 1~2mg of cobalt chloride, distilled water
1000mL, pH 7.0.
2. enriched medium：Distilled water 1000mL, 5.0~10g of peptone, beef extract 2~5g, NaCl 5~15g, pH
7.2。
Embodiment 1
Staphylococcus pasteuri (Staphylococcus pasteuri) KY-06 screening and identification.
The cud and ruminant stomach contents samples for the ox that experiment material is collected from Tianjin slaughterhouse.
Take sample diluting liquid (extension rate 10-6) 0.1ml, it is coated on screening and culturing medium flat board, in 30 DEG C of insulating boxs
Culture, picking single bacterium colony is seeded to screening and culturing medium, and more than hot housing 48h repeats line purifying 3 times, obtains purifying bacterial strain；
Identified by 16S rDNA and Biolog microorganism classifications identification systems, be defined as Staphylococcus pasteuri, systematic evolution tree such as Fig. 1
It is shown, identify that sequencing result is such as by 16S rDNA and Biolog microorganism classifications identification systems<400>Sequence shown in 1, i.e.,：
5’-AAAGTGGAATTCCATGTGTAGCGGTGAAATGCGCAGAGATATGGAGGAACACCAGTGGCGAAGGCG
ACTTTCTGGTCTGTAACTGACGCTGATGTGCGAAAGCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGC
CGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGG
GGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCG
AAGCAACGCGAAGAACCTTACCAAATCTTGACATCCTTTGACCGCTC-3’。
Embodiment 2
Drops of Staphylococcus pasteuri (Staphylococcus pasteuri) KY-06 to the macromolecular organic matter such as cellulose
Solution experiment.
Deal with objects as screening and culturing medium, the method for operation is shake flask culture.Specific implementation step is as follows：Embodiment 1
The Staphylococcus pasteuri KY-06 is seeded to enriched medium, 30 DEG C of shaken cultivations to increased logarithmic phase (bacterium dense about 108Individual/
Ml), it is inoculated in 3% inoculum concentration in liquid screening medium, 30 DEG C of shaken cultivations 1 day, maintenance dissolved oxygen is 1.0~3.0ppm；
Centrifuging and taking supernatant determines the strains for degrading cellulose etc. as crude enzyme liquid, according to DNS colorimetric methods (GB/T 23881-2009) and divided greatly
Sub- complexity organic matter produces the amount of reduced sugar.As a result show, per mL, the bacterial strain crude enzyme liquid can averagely produce the μ g/ of reduced sugar 618.33
min。
Embodiment 3
Synergy of Staphylococcus pasteuri (Staphylococcus pasteuri) KY-06 to municipal sludge anaerobic digestion is real
Test.
Deal with objects as anaerobism municipal sludge, anaerobic reaction system running is simulated using 400mL.Specific implementation step is such as
Under：Staphylococcus pasteuri KY-06 described in embodiment 1 is seeded to enriched medium, and (bacterium is dense to increased logarithmic phase for 30 DEG C of shaken cultivations
About 108Individual/ml) it is inoculated in 0.2% inoculum concentration in simulation anaerobic reaction system, transported under middle temperature (35 ± 2 DEG C) anaerobic condition
OK.As shown in Fig. 2 being monitored to the gas producing efficiency of reaction system, and it is analyzed, as a result shows with non-inoculation group,
Within the reaction time of 300 hours, Staphylococcus pasteuri KY-06 can be such that mixing sludge gas production is averagely lifted compared with control group
15.77%, bio natural pond potential energy MP increase by 27.84%, it was demonstrated that the microbial inoculum is respectively provided with notable synergistic effect, and energy to anaerobic digestion
Effective substrate conversion efficiency of anaerobic digestion can directly be participated in by significantly improving in sludge, obtain the overall production natural pond potential energy of reaction system
Lifting, so as to reach gas production and the dual reinforcing effect of mud decrement.
The above-mentioned detailed description carried out with reference to embodiment to a kind of Staphylococcus pasteuri KY-06 with application, is illustrative
Rather than it is limited, several embodiments can be included according to limited scope, therefore do not departing from overall structure of the invention
Changing and modifications under thinking, should belong within protection scope of the present invention.
Claims (7)
1. a kind of Staphylococcus pasteuri KY-06, it is characterised in that：Its preserving number is CGMCC No.10173.
2. Staphylococcus pasteuri KY-06 according to claim 1, it is characterised in that：Thalline in spherical, a diameter of 0.5~
1.5 μm, Gram-positive, no gemma is identified by 16S rDNA and Biolog microorganism classifications identification systems, is defined as Pasteur
Staphylococcus Staphylococcus pasteuri, with nucleotide sequence shown in sequence in sequence table 1.
3. Staphylococcus pasteuri KY-06 according to claim 2, it is characterised in that：The nucleotide sequence belongs to bacterium
The V3 variable regions of 16S rDNA sequences.
4. Staphylococcus pasteuri KY-06 according to claim 1, it is characterised in that：It is isolated from the ruminant stomach of ox, in sieve
It is in faint yellow to select and bacterium colony is shown as on flat board, and neat in edge, surface is smooth, convex protuberance.
5. Staphylococcus pasteuri KY-06 according to claim 1, it is characterised in that：It has the degraded of sludge organic matter
Ability.
6. applications of the Staphylococcus pasteuri KY-06 in natural pond reaction synergy is produced to anaerobic digestion described in claim 1.
7. Staphylococcus pasteuri KY-06 according to claim 6 application, it is characterised in that：By the bacterial strain Pasteur Portugal
Grape coccus KY-06 is seeded in enriched medium, and 30 DEG C of shaken cultivations are to increased logarithmic phase, and bacteria concentration is 106~1010Individual/ml,
Centrifuging and taking bacterium solution supernatant, supernatant is determined to macromolecular organic matter according to DNS reduced sugar colorimetric National Standard Method GB/T 23881-2009
Catabolism ability, and be inoculated in 0.1~0.5% inoculum concentration in anaerobic digestion reaction system, realize the bacterial strain to detesting
The synergistic effect of oxygen digestion reaction.
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