CN114703093A - Facultative anaerobic complete denitrification aerogenic bacterium Y23 and application thereof - Google Patents
Facultative anaerobic complete denitrification aerogenic bacterium Y23 and application thereof Download PDFInfo
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- 241000894006 Bacteria Species 0.000 title claims abstract description 35
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- 241000589614 Pseudomonas stutzeri Species 0.000 claims abstract description 24
- 238000011084 recovery Methods 0.000 claims abstract description 22
- 239000010865 sewage Substances 0.000 claims abstract description 17
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000000034 method Methods 0.000 claims abstract description 11
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 10
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- 101710144092 Periplasmic nitrate reductase Proteins 0.000 description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 3
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- 238000001514 detection method Methods 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000011790 ferrous sulphate Substances 0.000 description 2
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- 238000011081 inoculation Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 239000001272 nitrous oxide Substances 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000004317 sodium nitrate Substances 0.000 description 2
- 235000010344 sodium nitrate Nutrition 0.000 description 2
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 2
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- 108091093088 Amplicon Proteins 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 108010025915 Nitrite Reductases Proteins 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
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- 239000001257 hydrogen Substances 0.000 description 1
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- 238000009630 liquid culture Methods 0.000 description 1
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 150000002823 nitrates Chemical class 0.000 description 1
- 108010028128 nitric-oxide reductase Proteins 0.000 description 1
- 108010076678 nitrous oxide reductase Proteins 0.000 description 1
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- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C09K8/00—Compositions for drilling of boreholes or wells; Compositions for treating boreholes or wells, e.g. for completion or for remedial operations
- C09K8/58—Compositions for enhanced recovery methods for obtaining hydrocarbons, i.e. for improving the mobility of the oil, e.g. displacing fluids
- C09K8/582—Compositions for enhanced recovery methods for obtaining hydrocarbons, i.e. for improving the mobility of the oil, e.g. displacing fluids characterised by the use of bacteria
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- E—FIXED CONSTRUCTIONS
- E21—EARTH OR ROCK DRILLING; MINING
- E21B—EARTH OR ROCK DRILLING; OBTAINING OIL, GAS, WATER, SOLUBLE OR MELTABLE MATERIALS OR A SLURRY OF MINERALS FROM WELLS
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- E21B43/16—Enhanced recovery methods for obtaining hydrocarbons
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- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
- C02F2101/163—Nitrates
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Abstract
The invention discloses pseudomonas stutzeri capable of carrying out complete denitrification under higher oxygen concentrationPseudomonas stutzeriY23 and its application are disclosed. The Pseudomonas stutzeri Y23 has been preserved in the China general microbiological culture Collection center (CGMCC) at 1 month and 5 days 2022, and the preservation number is CGMCC No. 24240. The pseudomonas stutzeri Y23 has complete denitrification path, and can react with nitrate under aerobic and anaerobic conditionsThe nitrogen is converted into nitrogen, and the method is suitable for the fields of nitrogen removal treatment of nitrogen-containing sewage, oil field microorganism improvement of crude oil recovery, prevention and control of oil reservoir acidification and pipeline corrosion caused by sulfate reducing bacteria and the like.
Description
Technical Field
The invention belongs to the field of microbial technology and environmental biotechnology, and relates to pseudomonas stutzeri capable of performing complete denitrification under higher oxygen concentrationPseudomonas stutzeriY23 and its application.
Background
Nitrate-reducing bacteria are a class of bacteria capable of reducing nitrate or nitrite to produce N2O and N2Is a general term for the bacterium (1). Nitrate reducing bacteria are anaerobic bacteria, and a small part of nitrate reducing bacteria are facultative anaerobic bacteria. Facultative anaerobic nitrate reducing bacteria preferentially perform aerobic respiration under aerobic environment to grow thallus quickly, and perform anaerobic respiration by using nitrate or nitrite as an electron acceptor to generate N under anaerobic condition2O and N2. At present, nitrate reducing bacteria are widely applied in the fields of sewage denitrification treatment and microbial oil recovery.
The denitrification of sewage by nitrate reducing bacteria is one of the main means of the denitrification treatment of sewage at present. The microbial denitrification is usually carried out under anoxic conditions, and the oxygen influences the microbial denitrification effect. Thus, NO can be converted at higher oxygen concentrations3-N in turn to NO2 -、NO、N2O and N2The complete denitrification microorganism has higher application value in the aspect of strengthening the sewage denitrification treatment. In addition, the nitrate reducing bacteria with complete denitrification capability can also reduce the greenhouse gas N to a certain extent2And (4) discharging O.
During microbial oil recovery, carbohydrates and nitrates are often injected into reservoir reservoirs to activate underground microbes for microbial enhanced oil recovery. On the other hand, nitrate reducing bacteria can preferentially obtain an electron donor (organic carbon source) compared with sulfate reducing bacteria, so that the nitrate reducing bacteria can inhibit the growth of the sulfate reducing bacteria through biological competition, and the problems of oil reservoir acidification, pipeline corrosion, personal safety and the like caused by hydrogen sulfide generated by metabolism of the sulfate reducing bacteria are solved. Nitrous oxide (N) produced by denitrifying bacteria2O) gas with CO2Has similar molecular structure and higher solubility than CO in nonpolar liquid2Can act like CO2Oil displacement (dissolving in crude oil to swell the crude oil and reduce the viscosity of the crude oil); n produced by denitrifying bacteria2Oil recovery can be enhanced by increasing reservoir pressure.
The complete denitrifying bacteria provided by the patent are separated from an oil reservoir in which microbial oil recovery is implemented, nitrate solution is injected into the oil reservoir for a long time to carry out microbial oil recovery, and nitrate reducing bacteria are the dominant microbial groups of the oil reservoir blocks. The strain can realize complete denitrification under higher oxygen concentration, and has higher application value in the fields of enhanced sewage denitrification treatment and enhanced microbial oil recovery.
Disclosure of Invention
One purpose of the invention is to provide a pseudomonas stutzeri capable of carrying out complete denitrification under higher oxygen concentrationPseudomonas stutzeriY23, the preservation number of the China general microbiological culture Collection center is CGMCC No. 24240.
The invention also aims to provide a bacterial suspension, fermentation liquor or a microbial inoculum containing the microbial strain and new application.
The invention provides the pseudomonas stutzeri capable of completely denitrifyingPseudomonas stutzeriY23, its bacterial suspension, its culture solution, its fermentation product, and the application of the microbial inoculum containing it in microbial oil recovery.
The invention also provides the complete denitrificationPseudomonas stutzeriPseudomonas stutzeriY23, bacterial suspension thereof, culture solution thereof, fermentation product thereof and application of microbial inoculum containing the same in preparation of microbial oil recovery products.
The invention provides the pseudomonas stutzeri capable of completely denitrifyingPseudomonas stutzeriY23, bacterial suspension thereof, culture solution thereof, fermentation product thereof and application of microbial inoculum containing the same in sewage treatment.
The invention also provides the pseudomonas stutzeri subjected to complete denitrificationPseudomonas stutzeriY23, bacterial suspension thereof, culture solution thereof, fermentation product thereof, and application of microbial inoculum containing the same in preparation of products or equipment for sewage treatment.
It is still another object of the present invention to provide a product containing the above-mentioned Pseudomonas stutzeri as an active ingredientPseudomonas stutzeriY23, or a bacterial suspension thereof, or a culture solution thereof, or a fermentation product thereof, or a microbial agent containing the same, wherein the product has any one of functions of microbial oil recovery and sewage treatment.
It is still another object of the present invention to provide a method for microbial oil recovery, comprising the following steps: in the process of oil extraction, the pseudomonas stutzeri is treatedPseudomonas stutzeriY23 fermentation liquor and nutrient are injected into the stratum from the oil sleeve annulus of the oil well, the anaerobic metabolism of the strain is used for producing gas to improve the yield of a single well, and simultaneously, the acidification of the oil reservoir caused by sulfate reducing bacteria is prevented and controlled; the pseudomonas stutzeri is treatedPseudomonas stutzeriY23 fermentation liquor and nutrient are injected into underground from water injection well, and the bacteria is used to produce gas under underground metabolism to increase crude oil recovery ratio and prevent and control oil deposit acidification caused by sulfate reducing bacteria.
A final object of the present invention is to provide a method for treating wastewater, comprising the steps of: in the process of sewage treatment, the pseudomonas stutzeri is addedPseudomonas stutzeriY23 bacterial suspension, bacterial agent or pseudomonas stutzeri coated on fillerPseudomonas stutzeri Y23。
Drawings
FIG. 1 shows a phylogenetic tree of strain Y23 constructed based on the 16S rRNA gene and its homologous sequence.
FIG. 2 shows the denitrification effect of strain Y23 in a sequencing batch bioreactor.
FIG. 3 shows the gas production of strain Y23 under different carbon sources.
FIG. 4 shows the gas components produced by strain Y23 and their relative contents.
Detailed Description
Example 1: isolation and characterization of Strain Y23
Strain Y23 was isolated from oil field produced fluids. Nitrate solution is injected into an oil reservoir of the oil field for a long time to carry out microbial oil recovery, and nitrate reducing bacteria are one of the dominant microbial groups of the oil reservoir. The specific steps of strain isolation are as follows: inoculating 10mL of oilfield produced liquid into a 500mL sterile plastic bottle containing 200mL of sterile denitrifying aerogenic bacteria enrichment culture medium, removing redundant air, screwing a bottle cap, and culturing in a constant temperature box until gas is generated. The ingredients of the aerogenic denitrifying bacteria enrichment medium are as follows: 0-1 g of disodium hydrogen phosphate, 0-1 g of potassium dihydrogen phosphate, 0-10 g of sodium nitrate, 0-1 g of ferrous sulfate, 0-1 g of magnesium sulfate hexahydrate, 0.01 g of calcium chloride, 0.01 g of yeast extract powder, 0-10 g of glucose and distilled water, adjusting the pH to 6-8, and sterilizing for 20min by high-pressure steam at 121 ℃. And (3) inoculating 5mL of enrichment culture solution to the enrichment culture system of the aerogenic denitrifying bacteria again, and culturing in a constant temperature box until no gas is generated. And (3) separating single bacteria by adopting a dilution coating method. And (3) verifying the gas production of a single strain, and selecting the strain with the maximum gas production, namely Y23.
The genome of the strain is extracted using a genome extraction kit, the strain 16S rRNA gene is obtained by PCR amplification using primers 27f (5 '-AGA GTT TGA TCC TGG CTC AG-3') and 1492r (5 '-GGT TAC CTT GTTACG ACT T-3'), and the obtained PCR product is sequenced to obtain 16S rRNA sequence information. The nucleotide sequence of the 16S rRNA of the Y23 strain was submitted to GenBank, and the results of comparison by BLAST search and homology analysis showed that the 16S rDNA sequence of the Y23 strain was found to be identical to that of known strainsPseudomonas stutzeriThe similarity of the strain is 99 percent, which indicates that the strain is pseudomonas stutzeri, and the strain is named as pseudomonas stutzeri by combining the identification resultsPseudomonas stutzeriAnd Y23. Based on the nucleotide sequence of the 16S rRNA of strain Y23, using the software MEGA constructA phylogenetic tree is built as shown in FIG. 1.
The strain has been preserved in China general microbiological culture Collection center (CGMCC for short, the address: No. 3 of West Lu No. 1 of the Beijing university Hokko Chaoyang district, the microbiological research institute of Chinese academy of sciences, zip code 100101) on 1 month and 5 days 2022, and the preservation number is CGMCC No. 24240.
Example 2: strain Y23 denitrification gene detection
Using the bacterial strain genome as a template, and using the denitrifying gene specific primer and the amplification condition which are disclosed in the academic paper to perform PCR amplification to obtain the bacterial strain denitrifying gene sequence fragment. The obtained PCR product is checked by 1% agarose gel electrophoresis and sent to Beijing Okkomy biotechnology Limited to carry out sequencing after no error. The sequence obtained by sequencing is submitted to NCBI database for BLAST analysis to determine the functional gene most similar to the sequence.
Sequencing analysis of denitrifying gene PCR amplicon shows that the strain Y23 has coding genes of Membrane-bound nitrate reductase (Nar), Periplasmic nitrate reductase (Nap), Nitrite reductase (Nir), Nitric oxide reductase (NorB) and Nitrous oxide reductase (Nos) related to the denitrifying process, and can convert NO into the product3-N in turn to NO2 -、NO、N2O and N2. Thus, strain Y23 is a denitrifying bacterium with complete denitrification. The membrane-bound nitrate reductase Nar is sensitive to oxygen molecules and plays a main role in the biological denitrification of anaerobic denitrification. The periplasmic nitrate reductase Nap is located in the periplasm of the cell and is insensitive to oxygen molecules. The existence of the gene encoding periplasmic nitrate reductase means that the strain Y23 can perform denitrification under aerobic conditions, namely, oxygen and nitrate are simultaneously used as electron acceptors for energy-producing metabolism.
Example 3: denitrifying denitrification effect of strain Y23 in sequencing batch bioreactor
Inoculating the Y23 strain into 50 ml LB liquid culture medium for overnight culture; centrifuging the culture solution at 4000r/min for 5min, and removing supernatantLiquid; suspending thalli which are settled to the bottom of a centrifuge tube by using a sterile denitrification culture medium, centrifuging for 5min at 4000r/min, pouring out supernatant, and adding 50 mL of the sterile denitrification culture medium to suspend the thalli; the strain Y23 was inoculated in a sequencing batch bioreactor at an inoculum size of 1%, and the reactor was drained of treated wastewater every 12 hours while supplementing an equivalent amount of wastewater (12 hours of hydraulic retention time). Simulated wastewater containing C4H4Na2O4·6H2O、NaNO3、KH2PO4、MnSO4·7H2O and FeSO4·7H2And (O). And (3) sampling every 30 minutes to detect the content of nitrate nitrogen and nitrite nitrogen in the treatment solution on the 24 th day when the reactor runs stably, and determining the change of pH value.
As shown in figure 2, the strain Y23 has good denitrification effect in the sequencing batch bioreactor, the degradation of nitrate nitrogen by the strain is mainly concentrated in the first 4 hours, the denitrification efficiency is close to 100%, no nitrite is accumulated basically in the process, and the pH value of the treated wastewater is increased to a certain extent.
Example 4: gas production effect of strain Y23 under different carbon sources
In carbon source experiments, the carbon sources commonly used in industry, namely sugars (glucose, sucrose and molasses), organic acids, small molecular alcohols (glycerol), petroleum hydrocarbons and the like are used as unique carbon sources, and the gas production rate of the Y23 strain under different carbon sources is researched. The dosage of each carbon source is 0-5 g/L of glucose, 0-5 g/L of glycerol, 0-5 g/L of sodium succinate, 0-5 g/L of sodium acetate, 0-5 g/L of sucrose and 0-5 ml/L of toluene.
As shown in FIG. 3, the strain can produce gas in a liquid culture medium with glucose, sodium succinate, glycerol, sodium acetate, molasses and sucrose as the only carbon sources, but the gas production rate has a large difference under different carbon source substrates. In the culture solution taking saccharides (molasses, glucose and sucrose) as carbon sources, the gas yield of the strain is the highest; in the culture solution with glycerol, sodium succinate and sodium acetate as carbon sources, the gas production effect is reduced. Therefore, from the application point of view, molasses can be used as the best carbon source for gas production of the strain Y23.
Example 5: analysis of gas component produced by Denitrification of Strain Y23
Inoculating the seed liquid into a 1L feeding bottle containing a 1L denitrification gas production culture medium according to the inoculation amount of 2%, screwing a bottle cap, reserving two-way interfaces on the bottle cap, and connecting a gas collecting bag (Bickmann organisms) at each interface. In order to eliminate the influence of air on the analysis of gas components generated by the strains, in the early culture stage, an air inlet valve of one air collecting bag is opened, when the gas collection amount reaches about 200ml, the air inlet valve of the air collecting bag is closed, and an air inlet valve of the other air collecting bag is opened for gas collection. The gas analysis adopts an Agilent 7890A gas chromatograph, the detectors are a Thermal Conductivity Detector (TCD) and a hydrogen Flame Ionization Detector (FID), and the detection limit is 1 ppm.
As shown in FIG. 4, the gas generated by the aerogenic denitrifying bacteria Y23 mainly comprises nitrogen, nitrous oxide and carbon dioxide. N is a radical of2Content up to 56%, N produced2Oil recovery can be enhanced by increasing reservoir pressure. N is a radical of2The content of O reaches 32 percent, and the gas and CO2Has similar molecular structure and higher solubility than CO in nonpolar liquid2Can act like CO2Oil displacement. CO 22The content of the oil displacement agent reaches 9 percent and is far higher than the concentration (0.03 to 0.04 percent) of carbon dioxide in the air, the oil displacement agent is mainly generated by the respiration of microorganisms, and the oil displacement agent can reduce the viscosity of crude oil after being dissolved in the crude oil to play a role in oil displacement.
Example 6: preparation of strain Y23 fermentation liquor and bacterial suspension
The first preparation method of the fermentation liquor comprises the following steps: inoculating the strain Y23 into LB liquid medium, standing or shake culturing on 120 rpm shaking table under the culture condition of 16-45 deg.C for 12-36h, and the bacterial concentration reaches 10 after culturing9-1010one/mL fermentation broth.
And a second fermentation liquor preparation method comprises the following steps: inoculating the strain Y23 into LB liquid culture medium, and shake-culturing on a shaker at 120 rpm to logarithmic phase; inoculating to sterile fermentation medium (containing disodium hydrogen phosphate 0-1 g, potassium dihydrogen phosphate 0-1 g, sodium nitrate 0-10 g, ferrous sulfate 0-1 g, magnesium sulfate hexahydrate 0-1 g, calcium chloride 0.01 g, yeast extract powder 0.01 g, and molasses 0-10 g) at 5% inoculation amount, and culturing at 16-45 deg.CCulturing for 12-36h, and dissolving oxygen amount of 0-10 mg/L; the bacterial concentration reaches 10 after the culture is finished9one/mL fermentation broth.
Preparation of bacterial suspension of strain Y23: diluting the bacterial liquid prepared by the first fermentation liquid preparation method and the second fermentation liquid preparation method according to a certain proportion to obtain bacterial suspension with corresponding concentration; or the bacteria liquid prepared by the first fermentation liquid preparation method and the second fermentation liquid preparation method is filtered or centrifuged to collect bacteria, and the bacteria is added into the corresponding solution according to a certain proportion to obtain the bacterial suspension with corresponding concentration.
Claims (8)
1. Pseudomonas stutzeri capable of performing complete denitrification nitrogen removal under higher oxygen concentrationPseudomonas stutzeriY23, wherein the preservation number of the microorganism is CGMCC No. 24240.
2. Pseudomonas stutzeri according to claim 1Pseudomonas stutzeriY23, its culture solution, its bacterial suspension or its bacterial agent are used in microbial oil recovery or in preparing microbial oil recovery products.
3. Pseudomonas stutzeri according to claim 1Pseudomonas stutzeriThe application of Y23, the culture solution thereof, the bacterial suspension thereof or the microbial inoculum thereof in sewage treatment or in the preparation of products or equipment for sewage treatment.
4. A product containing Pseudomonas stutzeri as an active ingredient according to claim 1Pseudomonas stutzeriY23 or its bacterial suspension, and the product has the functions of any one of microbial oil recovery and sewage treatment.
5. A product containing Pseudomonas stutzeri as an active ingredient according to claim 1Pseudomonas stutzeriY23, and the product has any one of functions of microbial oil recovery and sewage treatment.
6. A product containing as an active ingredient the pseudoschwertmann as defined in claim 1MonomonasPseudomonas stutzeriY23, said product having any one of the functions of microbial oil recovery and sewage treatment.
7. A method of microbial enhanced oil recovery, comprising the steps of: a process for recovering oil, which comprises using the Pseudomonas stutzeri of claim 1Pseudomonas stutzeriY23 fermentation liquor and nutrient are injected into the stratum from the oil casing annulus of the oil well, the anaerobic metabolism of the strain is utilized to produce gas, the yield of a single well is improved, and simultaneously, the acidification of an oil reservoir caused by sulfate reducing bacteria is prevented and controlled; alternatively, the Pseudomonas stutzeri of claim 1Pseudomonas stutzeriY23 fermentation liquor and nutrient are injected into underground from water injection well, and the bacteria is used to produce gas under underground metabolism to increase crude oil recovery ratio and prevent and control oil deposit acidification caused by sulfate reducing bacteria.
8. A sewage treatment method is characterized by comprising the following steps: in the process of sewage treatment, the pseudomonas stutzeri as claimed in claim 1 is addedPseudomonas stutzeriY23 bacterial suspension, bacterial agent or pseudomonas stutzeri coated on fillerPseudomonas stutzeri Y23。
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