KR101008568B1 - Novel Aquitalea sp. 5YN1-3 strain for effective denitrification - Google Patents

Abstract

The present invention relates to a low nutrient novel bacterium Aquitalea sp. 5YN1-3 strain having excellent nitrate removal function isolated from the foliar soils of Yong swamp in Korea and a method using the characteristic nitrogen removal function of the strain. to be.

Description

Novel Aquitalea sp. 5YN1-3 strain for effective denitrification

The present invention provides a novel bacterium Aquitalea sp. 5YN1-3 , which has excellent nitrate removal activity and produces various nitrate scavenging enzymes among low-nutrition bacteria isolated from larvae soil . It is about method to use for ecosystem purification.

The biological removal method applied to the conventional nitrogen removal is 1) biological filtration process using activated carbon as a medium, and 2) ammonia nitrogen is oxidized to nitrate nitrogen using aerobic nitrification bacteria and returned to anoxic conditions to recover nitrate nitrogen. There is a nitrification-denitrification process that reduces and denitrates to nitrogen gas.

Accumulation of organic nitrogen is an important environmental problem. Organic nitrogen is accumulated in soil by nitrogen fertilizer, which is continuously administered to soil for food production, and livestock wastewater generated by livestock raising. The accumulation of nitrogen above the self-cleaning capacity of the ecosystem directly affects the acidification of the soil and eutrophication of the water quality. In addition, when nitrogen is accumulated in the environment, poor soil conditions are created, and a lack of nitrogen decomposers in the soil leads to a vicious cycle of nitrogen accumulation. Nitrogen accumulates in the environment in the form of organic nitrogen (Org-N), ammonia nitrogen (NH ₃ -N), nitrite nitrogen (NO ₂ -N) and nitrate nitrogen (NO ₃ -N). The main forms of nitrogen remaining in are organic nitrogen and ammonia nitrogen. In order to completely remove nitrogen accumulated in the environment, both nitrification and denitrification of nitrogen must be carried out. Nitrogen contamination is primarily present as ammonia nitrogen, but as the contamination is restored, it is oxidized back to nitrate nitrogen in an aerobic state. This oxidation process is called nitrification. The main products of nitrification by oxidation are in the form of nitrates (NO _3- ) and nitrites (NO _2- ). In the natural ecosystem nitrogen cycle, these two nitrification substances are finally reduced to nitrogen gas (N ₂ ) and returned to the air. After all, if the above two nitrification products are not removed quickly, the excess accumulation of nitrogen is not solved.

The denitrification process involves a series of chain enzymes as shown in the diagram below. In nature, by bacteria such as Nitrobacter or Nitrocystis,

^{_{NO 2 - + (1/2) O}} 2 ↔ NO 3 -

When these nitrates accumulate in the soil or environment, they are converted into nitrogen by the anaerobic denitrification mechanism below by various microorganisms in nature.

2NO ₃ + 10H (organic _{origin) → N 2 ↑ + 4H 2} O + 2OH - ( or)

2NO ₂ + 6H (organic _{origin) → N 2 ↑ + 2H 2} O + 2OH -

Enzymes involved in the process of the final release of nitrates into the environment in the form of nitrogen is required of enzymes or isozymes corresponding to the following three steps as reductases.

(Step 1) NO ₃ ⁻ → NO ₂ ⁻ ; Nitrate reductase ( narG gene)

(Step 2) NO ₂ ⁻ →→→ NO → N ₂ O; Nitric oxide reductase ( norB gene)

(Step 3) N ₂ O → N ₂ ↑; Nitrous oxide reductase ( nosZ gene)

Therefore, if the step enzyme to be finally reduced to nitrogen after the removal of nitrate is not present in the microorganism is incomplete denitrification does not eliminate the nitrogen accumulation.

Nitrate products in the environment cannot be removed by natural nitrogen cycles if they accumulate in excess. The reason is that the microbial community in the soil itself changes the dominant species depending on the acidity and nitrate concentration in the soil. Nitrogen, once accumulated in the soil, changes the healthy dominant species in the microbial community, resulting in a poor circulation of nitrogen, resulting in a vicious cycle of nitrogen accumulation.

Means to solve this problem are largely chemical and biological methods. Chemical methods include the use of catalysts to absorb nitrogen, but large-scale processes can exacerbate environmental pollution, making biological methods economical and reducing energy consumption. Among the special soil environment microorganisms in Korea, microorganisms should be selected by setting some prerequisites for the application in the site of actual nitrogen overaccumulation. 1) a new environmental microorganism belonging to a genus that has never been industrialized; 5) The removal rate was selected based on the bacterium, which was faster and more efficient than the comparison group, and 5) the bacterium which was able to remove the nitrate even with a small amount of bacteria.

The present invention describes the denitrification function of the new Aquitalea bacterium through various assays. In addition, this strain is a new bacterium isolated from the swamp, which is a very special ecosystem in Korea, and it is expected that novelty of genes and enzymes related to the denitrification function of the bacterium is expected. It can be usefully used. In particular, there are no cases of patenting the denitrogen effect in the genus to which this strain belongs.

The present invention isolates a new strain Aquitalea sp. 5YN1-3 having excellent denitrogen function, utilizes the denitrification function of the bacterium itself, and improves denitrification enzyme-related genes and enzymes present in the bacterium. It aims to provide a material for removal of excess nitrogen accumulated in the ecological environment.

One aspect of the present invention provides Aquitalea sp. 5YN1-3 strain KACC91389P with excellent denitrogen function.

In addition, another aspect of the present invention provides a method for denitrifying soil or water quality using Aquitalea sp. 5YN1-3 strain KACC91389P.

The low-nutrition bacterium Aquitalea sp. 5YN1-3 strain according to the present invention showed functional characteristics satisfying all of the above prerequisites, and a series of denitrogenases involved in effectively removing nitrate excess accumulation with nitrogen gas. All enzymes were produced. The Aquitalea sp. 5YN1-3 strain is a low-nutrition strain that has not yet been reported at home and abroad. New low-nutrition bacteria play an important role in the soil environment, but are not useful in high-nutrient media at normal concentrations, which is useful for the search for new bioactive functions.

In addition, the Aquitalea sp. 5YN1-3 of the present invention is a bacterium isolated from a swamp, which is a special ecological environment in Korea, and can be expected to have uniqueness of denitrification-related genes and enzymes. Industrially, the strain itself or its products can be utilized in an overdose of nitrogen fertilizers or nitrogen organics, which can be used for denitrification processes to improve the agricultural environment where the C / N ratio is imbalanced. In particular, this strain is a new microorganism that has emerged as a new species, and Aquitalea sp. (Genus), to which it belongs, has only one standard strain and there are no cases of patenting the denitrogen effect of related strains at home and abroad. .

The present invention is composed of excellent nitrate removal function and denitrification-related industrial enzymes of Aquitalea sp. Isolation of this strain is not possible with conventional methods, but after isolation the proliferation can be cultured by conventional methods.

Hereinafter, the strain isolation, species classification, biochemical identification, isolation of related genes, and denitrogenase assay of the present invention will be described by examples.

Example 1 Strain Selection

Soil samples for bacterial isolation were collected from the deciduous soils of forest areas in the "arm swamp" (GPS coordinate; 38 ° 12'53 "N 128 ° 07'30" E) adjacent to the unarmed area. The collected sample was removed by a 2mm sieve, and then shaken at 150 rpm for 30 minutes in 0.85% NaCl solution to prepare a series of diluted suspensions (10 ^2-5 ). Suspensions were taken in 1/10 milliliters of each dilution series and used as a low-nutrient medium for R2A solid medium (Yeast extract; 0.5 g / L), meat peptone (0.5 g / L), and casamino acid (0.5). g / L), glucose (0.5 g / L), starch (0.5 g / L), dipotassium hydrogen phosphate (0.3 g / L), MgSO ₄ (0.05 g / L), sodium pyruvate; 0.3 g / L), 1.5% agar, pH 7.0-7.2) and incubated in a thermostat controlled at 28 ℃. Colonies that appeared after 5 days of incubation were separated by repeated repeated 2-3 times with platinum. From this, a total of 600 strains were isolated, and the denitrification function was explored using these strains as test strains.

28 ° C in sterilized Durham tube and liquid denitrogen test medium (Beef extract (3g / L), peptone (5g / L), KNO ₃ (0.5g / L)) , Suspended culture for 1 day was consumed oxygen dissolved in the medium. After the strain was grown properly, about 1 milliliter of sterilized paraffin oil was applied to the medium and allowed to stand at 28 ° C. for 2 days while blocking oxygen in the air. The denitrification function was determined by the gas bubbles trapped in the tube, in addition to the conversion of nitrate (NO ₃ −) from the medium to nitrogen (N ₂ ) gas in an electron transfer system. 5YN1-3, which is weak green among the disclosed strains, is a gram-negative bacillus, which is a low-nutritic bacterium that forms viscous colonies on R2A solid medium. Oxygen demand is aerobic and has a facultative anaerobic nature that grows even in weak anaerobic conditions.

Example 2 Molecular Biological Species Analysis of 5YN1-3

16S rRNA gene sequencing was used to identify 5YN1-3 bacteria. DNA extraction of the isolate strain with a DNA extraction kit (Toyobo, Japan) followed by universal primer 27F (5'-AGAGTTTGATCMTGGCTCAG-3 ') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3') (Schmidt et (1991) Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing.J Bacteriol 173: 4371-4378) was used to amplify the 16S rDNA gene. The PCR product thus obtained was reacted using a DNA sequencing knit (DNA sequencing knit; BigDye terminator Cycle Sequencing Ready Reacions v. 3. Sequence (SEQ ID NO: 1) was analyzed. This is shown in FIG.

Example 3 Species Classification and Deposit of 5YN1-3 Strains

5YN1-3 strains were classified based on the 16S rDNA sequence of FIG. Figure 2 is a lineage classification according to 16S rDNA of the microorganism 5YN1-3 strain of the present invention. 16S rDNA sequences were identified to species through the BLAST program of the US NCBI server. ClustalW algorithm (Thompson et al. (1994) to improve the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.Nucleic Acids The base sequence was aligned using Res 22: 4673-4680). The sorted datasets were developed using the MEGA version 4 (Tamura et al. (2007). MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24: 1596-1599) program. The stability of the branch (bootstrap value) was investigated through 1,000 resampling. 5YN1-3 was 98.6% identical to Aquitalea magnusonii , the closest standard monarch, by phylogenetic similarity analysis using 16S rDNA sequences. Only one species of the genus Aquitalea is known throughout the world so far (Lau et al. (2006). Aquitalea magnusonii gen.nov ., Sp.nov ., A novel Gram-negative bacterium isolated from a humic lake.Int J Syst Evol Microbiol 56: 867-871). According to the above results, the present inventors have found that the microbial strain of the present invention is in the genus Aquiltalea . The strain belonging to the strain Aquitalea sp. ( Aquitalea sp.) 5YN1-3 strain was deposited on June 26, 2008 at the Rural Development Administration Agricultural Microbial Resources Center and received accession number (KACC 91389P).

Example 4 Enzyme Characterization Using API Kit of Aquitalea sp. 5YN1-3 Strain

To investigate the biochemical properties of the Aquitalea sp. 5YN1-3 strain, the strain was incubated at 28 ° C. for 24 hours, and a single colony was collected and suspended in 0.85% NaCl buffer. The cultured bacteria were examined for their biochemical properties using various API kits (20NE, 32GN; BioMerieux Inc.) after adjusting the OD values based on standard buffers. In the analysis, the standard strain Aquitalea magnusonii was used as a control. The results are shown in Table 1.

As shown in Table 1, Aquitalea sp. 5YN1-3 has a significant difference in biochemical properties from standard strains in assimilation and fermentation. Not only the sequencing but also the biochemical properties of the new species can be seen as supporting evidence.

Table 1 Comparison of Biochemical Enzyme Characteristics of Aquitalea sp. 5YN1-3 and Standard Strains

Physiological and chemical features 5YN1-3 ^* Standard strain ^* Growth temperature (℃) 10-40 28-37 DNA G + C Volume (% Molecular) 56 59.2 Glucose Fermentation - + Urease - + Gelatin hydrolysis - (+) Assimilation   Sodium malate - +   Salinity - + Enzyme reaction   Alkaline phosphatase - +   Esterase lipase (C8) - +   Naphthol-AS-BI-phosphohydrolase - +

* "+" Indicates a positive reaction, "-" indicates a negative reaction, and "(+)" indicates a weak positive reaction.

Enzyme biochemical characteristics which are positive in both 5YN1-3 and standard strains not shown in Table 1 are as follows. Catalase, Oxidase, Indole production, Arginine dihydrolase. In addition, D -glucose, N -acetylglucosamine, potassium gluconate, capric acid, malic acid, trisodium citrate, D -ribose, inositol, sodium acetate, lactic acid, L -alanine, L -serine, propionic acid, valeric acid acid, L -histidine, 3-hydroxybutyric acid, L -proline, esterase (C4), leucine arylamidase, acid Acid phosphatase was positive in all.

Example 5 Lipid Characterization of Aquitalea sp. 5YN1-3

Fatty acid characterization is a method for identifying the physiological properties of bacteria. After culturing Aquitalea sp. 5YN1-3 and standard strains at 30 ° C. for 24 hours, lipid components of each bacterium were extracted. Extracted lipids were methylated and analyzed using standard gas chromatography Microbial Identification (MIDI) system (Sasser (1990) Identification of bacteria by gas chromatography of cellular fatty acids.Technical Note 101, DE: MIDI Inc.). The results are shown in Table 2.

As shown in Table 2, the standard strain and Aquitalea sp. 5YN1-3 are different species belonging to the same genus with different lipid properties.

[Table 2] Lipid Characteristics of Aquitalea sp. 5YN1-3 and Standard Strains

Fat acid 5YN1-3 Standard strain C _{10: 0} 3-OH 6.8 1.5 C _{12: 0} 9.0 5.1 C _{12: 0} 3-OH 4.0 1.2 C _{14: 0} 1.8 2.8 C _{15: 0} - 2.0 C _{16: 0} 25.6 21.7 C _{17: 1} Omega-6C - 1.2 C _{18: 1} Omega-7C 6.6 8.0 iso-C _{15: 0} 2-OH or
C _{16: 1} Omega 7C 45.5 52.5

* Analysis values below 0.5% were not detected.

Example 6 Denitrification Related Gene Verification of Aquitalea sp. 5YN1-3

In order to confirm the presence of the denitrogenase-related enzyme gene of Aquitalea sp. 5YN1-3, 5YN1-3 DNA was isolated and fragments of representative denitrogen genes were amplified by PCR. The primer pairs used are as follows for each gene.

Step 1) narG gene amplification;

TAYGTSGGGCAGGARAAACTG / CGTAGAAGAAGCTGGTGCTGTT

Step 2) norB gene amplification;

GACAAGNNNTACTGGTGGT / GAANCCCCANACNCCNGC

Step 3) NosZ Gene Amplification

CGYTGTTCMTCGACAGCCAG / CGSACCTTSTTGCCSTYGCG

PCR amplification was repeated amplified 30 times using (Tak polymerase) 1 unit (94 ℃, 1 minute) → (Primer Melting point-5 ℃, 30 seconds) → (72 ℃, 30 seconds) cycle. The amplified PCR product was cloned into a TOPO vector (Invitrogen Co.) to determine the base sequence as described above. The base sequence was changed into protein amino acid sequence using NCBI's BLASTX algorithm to search the database. The result is shown in FIG.

Figure 3 is a partial sequence comparison of the denitrification related genes of the Aquitalea sp. 5YN1-3 strain, "Query sequence" is Aquitalea sp. 5YN1-3 Refers to the denitrification gene fragment nucleotide sequence amplified by PCR from the strain. GenBank Gene Identification (gi) numbers of the denitrification genes showing the highest similarity are also indicated. As shown in Figure 3, it was possible to detect the presence of a well-known enzyme gene in each step of denitrification.

Example 7 Verification of Nitrate Removal Efficiency of 5YN1-3

In order to confirm that the Aquitalea sp. 5YN1-3 strain converts the nitrogen component of the medium into nitrogen gas through metabolism, NO ₃ − of KNO ₃ is used as an acceptor of the electron transport system. Was used. Strain Aquitalea sp. 5YN1-3 was placed in sterile R2A liquid medium and cultured for one day. Full-grown by the active dilution good strain to be the initial OD ₆₀₀ absorbance of 0.01 in a nitrate medium (R2A + 100ppm KNO3), while stationary culture for 28 ℃, 8 sigan nitrate salts which remain in the culture medium (NO ₃ ^-), and nitrite ( NO ₂ ^-) ion concentration was measured. The nitrate medium was boiled in water for 5 minutes prior to adding the strain to remove dissolved oxygen. After the strain was added, sterilized paraffin oil was applied to about 1 milliliter and stationary cultured in the state of blocking oxygen in the air. The medium was sampled by time and ion chromatography was used to convert nitrate (NO ₃ ⁻ ) from the medium to nitrite (NO ₂ ⁻ ) into nitrogen (N ₂ ) gas in an electron transport system. The result is shown in FIG.

The solid line in FIG. 4 is Aquitalea sp. 5YN1-3 and the dotted line shows the denitrification pattern of the comparative strain Aquitalea magnusonii . In the anaerobic, oxygen-blocked state, Aquitalea sp. 5YN1-3 consumed all of the nitrate (NO ₃ ⁻ ) in the medium within 4 hours, and the resulting nitrite (NO ₂ ⁻ ) was nitrate (NO ₃ ^−). ) Peaked at 4 hours of total consumption. The resulting nitrite (NO ₂ ⁻ ) began to decrease in anaerobic conditions and consumed all of the initial nitrate present in the medium in a total of 6 hours. The final OD ₆₀₀ of the two bacteria growing little by little in the anaerobic state was the same (OD ₆₀₀ ~ 0.21). Comparative strains showed a similar pattern, but Aquitalea sp. 5YN1-3 showed excellent denitrification ability to remove all given nitrates in a short time of about 2 hours (Fig. 6).

Example 8 Validation of Denitrogen Capacity of Aquitalea sp. 5YN1-3

Multiple R2A denitrification media in which the denitrification capacity of the Aquitalea sp. 5YN1-3 strain was 5 to 100 times greater than the nitrate medium used in Example 7 (maximum NO ₃ ⁻ 10,000 ppm). Assay in the state. In the same experiments a day such that the passage before the activity is to remove the dissolved oxygen from the good bacteria for 10 minutes in boiling water in advance nitrate medium (each of the nitrate concentration of 500ppm, 1,000ppm, 5,000ppm, 10,000ppm) Initial OD ₆₀₀ ~ 0.01 in 10 ml of culture Inoculated. Sterilized paraffin oil was applied about 1 millimeter, and then maintained in an anaerobic state for 12 hours by standing culture. Re-measured OD ₆₀₀ after 12-hour anaerobic culture was the same at 0.2. Samples were taken at the same 12 hours and then the amount of nitrates (NO ₃ ⁻ ) and nitrites (NO ₂ ⁻ ) remaining in each denitrification medium was measured by ion chromatography. The result is shown in FIG.

As shown in FIG. 5, Aquitalea sp. 5YN1-3 remained only 20% or less of the initial nitrate in the anaerobic state, and all others were removed. On the other hand, as the standard nitrate concentration increased, the denitrification capacity decreased, and the denitrification capacity was remarkably decreased in the high nitrogen salt condition of about 10,000 ppm, and only about 10% of the total nitrate was removed. Therefore, at the same time, Aquitalea sp. 5YN1-3 in high nitrogen salt medium was found to have an excellent denitrification ability to convert nitrogen components into nitrogen gas up to 7 times more than standard strains.

As described above, the Aquitalea sp. 5YN1-3 strain of the present invention has the ability to produce a series of denitrogenases having excellent denitrogen function. Aquitalea sp. 5YN1-3 strains can be obtained in a conventional manner in sufficient amounts for industrial use in one day when mass cultivation. 4 and 5, Aquitalea sp. 5YN1-3 was able to obtain a sufficient denitrogen effect even in a small number of bacteria with an OD ₆₀₀ of 0.2. In other words, with proper tinting, Aquitalea sp. 5YN1-3 is capable of a greater amount of denitrogenation more efficiently at a given time. In particular, this property of the strain has not yet been reported at home and abroad. Low nutrient bacteria derived from Aquitalea sp. 5YN1-3 play an important role in soil environments, but are not useful in the normal concentration of high-nutrient media, which is useful for the search for new bioactive functions. In addition, the Aquitalea sp. 5YN1-3 of the present invention is a bacterium isolated from a swamp, which is a special ecological environment in Korea, and can be expected to have uniqueness of denitrification-related genes and enzymes. Compared to the existing denitrification microorganisms, the bacterium has the ability to grow well in water or wet areas. Therefore, long-term survival is possible even in areas with severe water pollution. Industrial applications are expected to be used in denitrification processes to improve agricultural environments where the fertilizer or nitrogen organics are overdosed using the strain itself or its products. In particular, because it has the entire group of denitrogen-related genes, it can be crushed and used for denitrogen using extracts of cells. There have been no cases of patenting the denitrogen effect in genus belonging to this genus.

1 is a complete sequence of 16S rDNA of the new Aquitalea sp. 5YN1-3 analyzed according to the present invention.

Figure 2 is a diagram showing a 16S rDNA lineage classification showing that the Aquitalea sp. 5YN1-3 strain of the present invention belongs to a new species.

3 is a BLASTX search result of a denitrogen-related gene sequence fragment of Aquitalea sp. 5YN1-3 of the present invention.

Figure 4 is a diagram showing the result of the ion chromatography analysis of the nitrate removal efficiency of the Aquitalea sp. 5YN1-3 strain of the present invention.

FIG. 5 shows that the Aquitalea sp. 5YN1-3 of the present invention has better denitrification ability per unit time under high nitrogen salt stress.

<110> Republic of Korea <120> Novel Aquitalea sp. 5YN1-3 strain for effective denitrification <160> 1 <170> KopatentIn 1.71 <210> 1 <211> 1406 <212> DNA <213> Aquitalea sp. <400> 1 catgcagtcg aacggtaaca gggagcttgc tccgctgacg agtggcgaac gggtgagtaa 60 tgcgtcggaa cgtgccgagt agtgggggat aactatccga aaggatagct aataccgcat 120 acgctttgag aaggaaagca ggggatcgca agaccttgcg ctattcgagc ggccgacgtc 180 tgattagcta gttggtgagg taaaggctca ccaaggcatc gatcagtagc gggtctgaga 240 ggatgatccg ccacactggg actgagacac ggcccagact cctacgggag gcagcagtgg 300 ggaattttgg acaatgggcg caagcctgat ccagccatgc cgcgtgtctg aagaaggcct 360 tcgggttgta aaggactttt gtcagggagg aaatccctaa ggttaatacc cttgggggat 420 gacagtacct gaagaataag caccggctaa ctacgtgcca gcagccgcgg taatacgtag 480 ggtgcaagcg ttaatcggaa ttactgggcg taaagcgtgc gcaggcggtt gtgtaagtct 540 gatgtgaaag ccccgggctc aacctgggaa ctgcattgga gactgcacgg ctagagtgcg 600 tcagaggggg gtagaattcc gcgtgtagca gtgaaatgcg tagagatgcg gaggaatacc 660 gatggcgaag gcagccccct gggatgacac tgacgctcat gcacgaaagc gtggggagca 720 aacaggatta gataccctgg tagtccacgc cctaaacgat gtcaactagc tgttgggggt 780 ttgaatcctt ggtagcgtag ctaacgcgag aagttgaccg cctggggagt acggccgcaa 840 ggttaaaact caaaggaatt gacggggacc cgcacaagcg gtggatgatg tggattaatt 900 cgatgcaacg cgaaaaacct tacctggtct tgacatgtac ggaacttgcc agagatggct 960 tggtgcccga aagggagccg taacacaggt gctgcatggc tgtcgtcagc tcgtgtcgtg 1020 agatgttggg ttaagtcccg caacgagcgc aacccttgcc attagttgct accatttagt 1080 tgagcactct aatgggactg ccggtgacaa accggaggaa ggtggggatg acgtcaagtc 1140 ctcatggccc ttatgaccag ggcttcacac gtcatacaat ggtcggtaca gagggtagcc 1200 aagccgcgag gtggagccaa tctcataaaa ccgatcgtag tccggatcgc actctgcaac 1260 tcgagtgcgt gaagtcggaa tcgctagtaa tcgcagatca gcatgctgcg gtgaatacgt 1320 tcccgggtct tgtacacacc gcccgtcaca ccatgggagt gagtttcacc agaagtgggt 1380 aggctaaccg taaggaggcc gctacc 1406  

Claims (2)

Aquitalea sp. 5YN1-3 strain KACC91389P with excellent denitrogen function. Aquitalea sp. 5YN1-3 strain KACC91389P for denitrification of soil or water quality.

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