CN114214258A - Mixed microbial inoculum and application thereof in removing methane and nitrate - Google Patents
Mixed microbial inoculum and application thereof in removing methane and nitrate Download PDFInfo
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- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 229910002651 NO3 Inorganic materials 0.000 title claims abstract description 34
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 title claims abstract description 34
- 239000002068 microbial inoculum Substances 0.000 title claims abstract description 23
- 241000894006 Bacteria Species 0.000 claims abstract description 13
- 241000862974 Hyphomicrobium Species 0.000 claims abstract description 7
- 241001467578 Microbacterium Species 0.000 claims abstract description 7
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 6
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- 238000002955 isolation Methods 0.000 description 2
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 2
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- JSPANIZMKMFECH-UHFFFAOYSA-L manganese(II) sulfate dihydrate Chemical compound O.O.[Mn+2].[O-]S([O-])(=O)=O JSPANIZMKMFECH-UHFFFAOYSA-L 0.000 description 1
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 1
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- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 1
- FDEIWTXVNPKYDL-UHFFFAOYSA-N sodium molybdate dihydrate Chemical compound O.O.[Na+].[Na+].[O-][Mo]([O-])(=O)=O FDEIWTXVNPKYDL-UHFFFAOYSA-N 0.000 description 1
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- B01D53/34—Chemical or biological purification of waste gases
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Abstract
The invention discloses a mixed microbial inoculum and application thereof in removing methane and nitrate, wherein the mixed microbial inoculum contains a methyl spore bacterium (Methylocystis sp) strain M9 and a Hyphomicrobium sp strain L1; in the mixed microbial inoculum, the viable bacteria content of the methylocyclosporium strain M9 and the raw silk micro-bacterium strain L1 respectively at least reaches 1.0 multiplied by 108CFU/mL, methylThe viable bacteria content ratio of the cyst strain M9 to the hyphomycete strain L1 is 1: 3-1: 15. The method combines the Methylocystis and the hyphomycete for controlling and repairing the pollution of methane and nitrate in the anoxic/anaerobic environment for the first time, and the mixed microbial inoculum has better methane and nitrate removing effect than that of a single microbial inoculum of Methylocystis sp, and has more engineering application value.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to a mixed microbial inoculum and application thereof in removing methane and/or nitrate.
Background
Methane is the second largest greenhouse gas next to carbon dioxide, but its greenhouse effect is about 25-28 times that of carbon dioxide. Methane is generated by the fermentation of organic matters by methanogens in an anaerobic environment and is released into the atmosphere, so that the biotechnology is the most economical and effective method for reducing methane emission. In particular, methane undergoes a wide range of anaerobic/anoxic zones during its release, and therefore, it is important to develop microbial agents that remove methane in an anoxic environment.
In addition, nitrogen pollution has been a pollution problem that plagues aqueous environments. Nitrogen pollution threatens human health and ecological balance while seriously destroying aquatic ecosystems. The nitrogen oxide is combined with rainwater to generate acid rain, and the acid rain enters a water body to cause acidification; discharge of industrial and domestic wastewater and loss of nitrogen fertilizer results in discharge of excessive nitrogen into the environmental water body, resulting in eutrophication of the water body and release of more methane from the sediment of the water body. Therefore, controlling nitrogen emissions in wastewater is an important issue that must be addressed.
Nitrate is one of the most common nitrogen sources in wastewater. The traditional method for removing nitrate nitrogen by denitrifying by using denitrifying bacteria and using organic matters as carbon sources can cause carbon source waste and increase the operation cost of wastewater treatment, and the method for removing nitrate by using methane as carbon sources reduces the emission of greenhouse gases while removing nitrate, treats waste by waste and can greatly improve economic benefits and environmental benefits.
The discovery of coupling nitrate reduction and methane oxidation in an anoxic/anaerobic environment provides a new technical approach for synchronously reducing greenhouse gas emission and nitrogen pollution. However, the currently widely studied functional microorganisms coupling nitrate reduction and anaerobic methane oxidation grow very slowly, the doubling time reaches 2 weeks or even more than 1 month, the removal efficiency of target pollutants is low, and the application of the method in the field of practical engineering is limited. Therefore, the prepared efficient mixed microbial inoculum capable of synchronously removing methane and nitrate is a key bottleneck problem in the engineering application of reducing and coupling nitrate with methane oxidation, and has great significance for the practical application of the efficient mixed microbial inoculum.
Disclosure of Invention
The invention aims to provide a mixed microbial inoculum capable of efficiently and synchronously removing methane and/or nitrate in a lack/anaerobic environment, reducing the methane discharge amount, reducing the greenhouse effect, effectively removing the nitrate in a water body, reducing the eutrophication of the water body and providing a new engineering technology application approach for synchronously reducing the methane and nitrate discharge.
The purpose of the invention is realized by the following technical scheme:
a mixed bacterial preparation contains Methylosporangium strain M9 and Micrococcus serinae strain L1;
the methyl ascomycete (Methylocystis sp) strain M9 is separated from a methane anaerobic oxidation reactor, and the colony is round, white, slightly raised, opaque, smooth in surface, wet and neat in edge; gram stain negative; the physiological and biochemical identification experiment results of the strain are as follows: citrate (positive), gelatin liquefaction (negative), H2S production (positive), methyl red test (negative), starch hydrolysis (positive), indole test (negative), catalase (positive), nitrate reduction (positive).
The 16S rDNA sequence of the strain M9 is shown in SEQ ID NO.1, and the classification position of the strain is closest to that of methyl ascomycete (Methylocystis sp.) through evolutionary tree analysis;
the strain M9 can be identified as Methylocystis sp by combining the above.
The filamentous microbe (Hyphomicrobium sp) strain L1 is obtained by separating from a methane anaerobic oxidation reactor, the colony of the filamentous microbe is brown and rod-shaped bacteria, the tip of the filamentous microbe is oval, and the filamentous microbe sprouts and breeds; gram stain negative; the physiological and biochemical identification experiment results of the strain are as follows: citrate (positive), gelatin liquefaction (negative), H2S production (negative), methyl red test (negative), starch hydrolysis (positive), indole test (negative), catalase (positive), nitrate reduction (positive).
The 16S rDNA sequence of the strain L1 is shown in SEQ ID NO.2, and the classification position is closest to the filamentous microbe (Hyphomicrobium sp.) through the evolutionary tree analysis;
the strain L1 can be identified as filamentous microbe (Hyphomicrobium sp.) by combining the above.
In the mixed microbial inoculum, the viable bacteria content of the methylocyclosporium strain M9 and the raw silk micro-bacterium strain L1 respectively at least reaches 1.0 multiplied by 108CFU/mL；
In the mixed microbial inoculum, the viable bacteria content ratio of the methylocyclosporium strain M9 to the raw silk micro-bacterium strain L1 is 1: 3-1: 15.
The mixed microbial inoculum can be used for removing methane and/or nitrate, and is particularly suitable for removing methane and nitrate in an anoxic/anaerobic environment;
the dosage forms of the mixed microbial inoculum comprise powder, solution and suspoemulsion.
Compared with the prior art, the invention has the following advantages and effects:
the method combines the Methylocystis sp and the Hyphomicrobium sp for controlling and repairing the pollution of methane and nitrate in the absence/anaerobic environment for the first time, and the mixed microbial inoculum has better methane and nitrate removing effect than a Methylocystis sp single microbial inoculum and has higher engineering application value.
Drawings
Fig. 1 is a phylogenetic tree of methyocystis sp.m 9.
FIG. 2 is a Hyphomicrobium sp.L1 phylogenetic tree.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Example 1: isolation and characterization of strains
1. Isolation of the Strain
Collecting 3mL of cement mixture in anaerobic reactor of river-south university B102, Xingdao Dong 855 in south village of Muyu district, Guangzhou province, centrifuging (4000rpm, 10min), collecting 1mL of supernatant, adding into 100mL of serum bottle containing 20mL of sterilized AMS liquid culture medium, culturing at 150rpm and 30 deg.C for two days, collecting 1mL of concentrate, and adding into 9mL of 0.01mol/L PBS buffer solution (sterilized) with pH of 7.2, wherein the PBS buffer solution mainly comprises KH2PO4、Na2HPO4NaCl and KCl are evenly blasted to obtain 10-2The concentration of (a) of (b),class pushing to 10 in turn-6Respectively sucking 30 mu L of bacterial suspension with each dilution multiple and the corresponding flat plate by using a sterile pipette, uniformly coating the bacterial suspension and the corresponding flat plate on an AMS solid culture medium by using a sterilized and cooled triangular coater, then inversely placing the coated flat plate in an anaerobic box, opening a small opening on an air bag filled with methane, then placing the air bag in the anaerobic box, and culturing for 14 days at the temperature of 30 ℃. After a single colony grows on the plate, the single colony is selected for multiple streaking purification, and a strain of bacteria is obtained by separation and stored on an inclined plane, which is numbered M9.
The AMS medium composition for screening for metysospora M9 contained (per liter): 0.1g MgSO4﹒7H2O，0.5g(NH4)Cl，0.56g KH2PO4，0.3g Na2HPO4，5mg CaCl，5mg FeSO4﹒7H2O, 1ml of trace element solution, agar 16-18g, pH 7.1.
The trace element solution comprises (per liter): 0.44g ZnSO4﹒7H2O，0.2g CuSO4﹒5H2O，0.17g MnSO4﹒2H2O，0.06g Na2MoO4﹒H2O，0.1g H3BO3，0.08g CoCl﹒6H2O。
Taking 3mL of the liquid sample (same as above) in the reactor (same as above), centrifuging (4000rpm, 10min), taking 1mL of supernatant and 9mL of PBS buffer solution (sterilizing), and washing uniformly to obtain 10-2The concentration of the bacterial suspension is sequentially similar to 10-6The bacterial suspension of each dilution of 30 mul is respectively absorbed into the corresponding plate by a sterile pipette, and is evenly coated on a screening solid culture medium (337 culture medium) by a triangular coater cooled after sterilization, and then the coated plate is placed in an anaerobic box upside down and cultured for 7 days at 30 ℃. After a single colony grows on the plate, the single colony is selected for multiple streaking purification, and a strain of bacteria is obtained by separation and stored on an inclined plane, which is numbered L1.
The "337" medium components used for screening Microbacterium filaginosum L1 include (per liter): 1.3g KH2PO4，1.13gNa2HPO4,0.5g(NH4)2SO4，0.2g MgSO4﹒7H2O，2.33mg CaCl，2.0g FeSO4﹒7H2O，1.0mg Na2MoO4﹒2H2O，0.67mg MnSO4﹒H2O，10ug CuCl2﹒2H2O，0.3％CH3OH, agar 16-18g, pH 6.8.
2. Identification of strains
(1) The physiological and biochemical identification results of the strains are shown in table 1:
TABLE 1 physiological and biochemical Properties of Strain M9 and Strain L1
| Item | M9 | L1 |
| Citric acid salt | + | + |
| Liquefaction of gelatin | - | - |
| H2S generation | + | - |
| Methyl Red test | - | - |
| Starch hydrolysis | + | + |
| Indole test | - | - |
| Contact enzyme | + | + |
| Nitrate reduction | + | + |
Note: "+" indicates positive reaction, and "-" indicates negative reaction
(2) 16S rDNA molecular characterization of M9 strain:
total bacterial DNA was extracted and the bacterial genome was PCR amplified using 16S rDNA universal primers (27F/1492R). After the PCR product is sequenced (finished by Huada gene), the sequencing result (SEQ ID NO.1) is subjected to homology comparison with 16S rDNA sequences of other strains reported in GenBank through NCBI blast, and related strains are selected for evolutionary tree analysis, as shown in figure 1, the classification position of the strains is closest to that of Methylocystis sp;
combined with its morphological characteristics and physiological and biochemical properties, it is most similar to Methylocystis sp. Bacterium M9 was therefore identified as Methylocystis sp and was designated Methylocystis M9.
(3) 16S rDNA molecular characterization of L1 strain:
total bacterial DNA was extracted and the bacterial genome was PCR amplified using 16S rDNA universal primers (27F/1492R). After the PCR product is sequenced (finished by Huada gene), the sequencing result (SEQ ID NO.2) is subjected to homology comparison with the 16S rDNA sequences of other strains reported in GenBank through NCBI blast, and related strains are selected for evolutionary tree analysis, as shown in figure 2, the classification position of the strains is closest to that of Hyphomicrobium sp);
combined with its morphological characteristics and physiological and biochemical properties, it is most similar to the genus Hyphomicrobium sp. Bacterium L1 was therefore identified as Hyphomicrobium sp and was designated Hyphomicrobium sp L1.
Example 2
Mixed bacterial agent and application thereof
(1) The strain M9 of the methyl spore fungus is inoculated into a serum bottle containing sterilized NMS liquid culture medium, high-purity methane is injected, the headspace methane concentration of an anaerobic bottle is 10% (v/v), the anaerobic bottle is activated in a shaker at the rotating speed of 150rpm/min at 30 ℃, the anaerobic bottle is cultured until the OD600 of the logarithmic growth phase is 0.5, and then the strain is inoculated into the culture medium according to the volume ratio of 5% for 7 days at 30 ℃. Collecting the enriched bacterial liquid, re-suspending in NMS culture medium, and obtaining colony number about 1.4X 10 by dilution plate counting method8CFU/mL。
(2) The strain Microbacterium filaginosum L1 is inoculated into a serum bottle containing sterilized '337' liquid culture medium, cultured in a seed culture medium at 30 ℃ for 3 days, and then inoculated into the culture medium at 5% by volume at 30 ℃ for 5 days. Collecting the enriched bacterial liquid, re-suspending the '337' culture medium, and obtaining the colony number of about 1.4X 10 by a dilution plate counting method8CFU/mL。
(3) And mixing the resuspended M9 bacterial liquid and the L1 bacterial liquid according to the ratio of 1: 5.
Example 3
Mixed bacterial agent and application thereof
Steps (1) and (2) were the same as in example 2;
(3) the resuspended M9 bacterial liquid and the L1 bacterial liquid are mixed according to the proportion of 1: 14.
Comparative example
The strain M9 of the methyl spore fungus is inoculated into a serum bottle containing sterilized NMS liquid culture medium, high-purity methane is injected, the headspace methane concentration of an anaerobic bottle is 10% (v/v), the anaerobic bottle is activated in a shaker at the rotating speed of 150rpm/min at 30 ℃, the anaerobic bottle is cultured until the OD600 of the logarithmic growth phase is 0.5, and then the strain is inoculated into the culture medium according to the volume ratio of 5% for 5 days at 30 ℃. Collecting the enriched bacterial liquid, re-suspending in NMS culture medium, and obtaining colony number about 1.4X 10 by dilution plate counting method8CFU/mL。
Respectively taking the bacterial liquid obtained finally in the examples 2 and 3 and the comparative example, inoculating the bacterial liquid into a serum bottle, wherein the culture solution in the serum bottle is sterilized NMS liquid culture medium, each of which is provided with 5 parallel cells, after sealing, flushing the serum bottle with argon gas to remove oxygen in the serum bottle, pumping a certain volume of gas after balancing the air pressure, injecting high-purity methane with the same content into the high-purity methane, wherein the methane concentration is about 220 mu mol, the nitrate nitrogen concentration is 8.0mg/L, placing the serum bottle at 30 ℃, oscillating at the rotating speed of 150rpm/min, culturing for 5 days, and comparing the effects of the product and a reference product on removing methane and nitrate in an anoxic environment in the culture period.
And (3) test results:
using the M9 bacterial suspension alone of the comparative example, the average consumption rate of methane was 6.57. mu. mol d-1And the average formation rate of carbon dioxide was 0.593. mu. mol d-1The removal rate of nitrate was 1.12mg L-1d-1。
By using the mixed bacterial liquid of the embodiment 2, the effect of the mixed bacterial agent M9+ L1 on removing methane and nitrate simultaneously in the absence/anaerobic environment is obviously better than that of M9, and the methane removal rate of the mixed bacterial agent M9+ L is 8.65 mu mol d-1The carbon dioxide generation rate was 0.851 and the nitrate removal rate was 1.18mg L-1d-1The methane degradation effect of the mixed microbial inoculum is 1.32 times that of the control, and the removal effect of the nitrate is about 1.05 times.
By using the mixed bacterial liquid of the embodiment 3, the effect of the mixed bacterial agent M9+ L1 on removing methane and nitrate simultaneously in the absence/anaerobic environment is obviously better than that of M9, and the methane removal rate of the mixed bacterial agent M9+ L is 13.2 mu mol d-1The rate of formation of carbon dioxide was 1.02 and the rate of removal of nitrate was 1.30mg L-1d-1The methane degradation effect of the mixed microbial inoculum is 2 times that of the control, and the nitrate removal effect is about 1.2 times.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Sequence listing
<110> river-south university
<120> mixed microbial inoculum and application thereof in removing methane and nitrate
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1355
<212> DNA
<213> Methylocystis sp
<220>
<223> 16S rDNA
<400> 1
tgcagtcgaa cgctgtagca atacagagtg gcagacgggt gagtaacgcg tgggaacgtg 60
cctttcggtt cggaataact cagggaaact tgagctaata ccggatacgc cctttggggg 120
aaagatttat tgccgaaaga tcggcccgcg tccgattagc tagttggtga ggtaatggct 180
caccaaggcg acgatcggta gctggtctga gaggatgatc agccacactg ggactgagac 240
acggcccaga ctcctacggg aggcagcagt ggggaatatt ggacaatggg cgcaagcctg 300
atccagccat gccgcgtgag tgatgaaggc cctagggttg taaagctctt tcgccaggga 360
cgataatgac ggtacctgga taagaagccc cggctaactt cgtgccagca gccgcggtaa 420
tacgaagggg gctagcgttg ttcggaatca ctgggcgtaa agcgcacgta ggcggatctt 480
taagtcaggg gtgaaatccc gaggctcaac ctcggaactg cctttgatac tggaggtctc 540
gagtccggga gaggtgagtg gaactgcgag tgtagaggtg aaattcgtag atattcgcaa 600
gaacaccagt ggcgaaggcg gctcactggc ccggaactga cgctgaggtg cgaaagcgtg 660
gggagcaaac aggattagat accctggtag tccacgccgt aaacgatgga tgctagccgt 720
tggggagctt gctcttcagt ggcgcagcta acgctttaag catcccgcct ggggagtacg 780
gtcgcaagat taaaactcaa aggaattgac gggggcccgc acaagcggtg gagcatgtgg 840
tttaattcga agcaacgcgc agaaccttac cagcttttga catgcccggt atgatcgcca 900
gagatggctt tcttcccgca aggggccgga gcacaggtgc tgcatggctg tcgtcagctc 960
gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa ccctcgccct tagttgccat 1020
cattaagttg ggcactctag ggggactgcc ggtgataagc cgcgaggaag gtggggatga 1080
cgtcaagtcc tcatggccct tacaggctgg gctacacacg tgctacaatg gcggtgacaa 1140
tgggaagcga aagggcgacc tggagcaaat ctcaaaaagc cgtctcagtt cggattgcac 1200
tctgcaactc gagtgcatga aggtggaatc gctagtaatc gcagatcagc acgctgcggt 1260
gaatacgttc ccgggccttg tacacaccgc ccgtcacacc atgggagttg gttttacccg 1320
aaggcgtttc gccaaccgca agggggcagg cgacc 1355
<210> 2
<211> 1395
<212> DNA
<213> Micromycelial sp (Hyphomicrobium sp.)
<220>
<223> 16S rDNA
<400> 2
gggaagtgcg gcgggcctaa acatgcaagt cgaacgctgt agcaatacag agtggcagac 60
gggtgagtaa cacgtgggaa tcttcctatc ggtacggaat agctcaggga aacttggggt 120
aataccgcat acgcccttcg ggggaaagat ttatcgccga tagatgagcc cgcgtctgat 180
tagctagttg gtgaggtaat ggctcaccaa ggcgacgatc agtagctggt ctgagaggat 240
gaccagccac actgggactg agacacggcc cagactccta cgggaggcag cagtggggaa 300
tattggacaa tgggcgcaag cctgatccag ccatgccgcg tgagtgacga aggtcttcgg 360
attgtaaagc tcttttggcg gggacgataa tgacggtacc cgcagaataa gccccggcta 420
acttcgtgcc agcagccgcg gtaatacgaa gggggctagc gttgttcgga attactgggc 480
gtaaagcgca cgtaggcgga ttgctaagtc aggggtgaaa tcccggggct caacctcgga 540
actgcctttg atactgacag tcttgagtcc ggaagaggtg agtggaattc ctagtgtaga 600
ggtgaaattc gtagatatta ggaagaacac cagtggcgaa ggcggctcac tggtccggta 660
ctgacgctga ggtgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg 720
ccgtaaacta tggatgctag ccgtcggcaa gcttgcttgt cggtggcgca gctaacgctt 780
taagcatccc gcctggggag tacggccgca aggttaaaac tcaaaggaat tgacgggggc 840
ccgcacaagc ggtggagcat gtggtttaat tcgacgcaac gcgaagaacc ttaccagctc 900
ttgacattta cggaccgttt ccagagatgg attcatccta gcaataggcc gtaggacagg 960
tgctgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg 1020
caaccctcgc cattagttgc catcatttag ttgggcactc tagtgggact gccggtgata 1080
agccggagga aggtggggat gacgtcaagt catcatggcc cttacgggct gggctacaca 1140
cgtgctacaa tggcggtgac aatgggcagc aacacagcaa tgtgaagcta atctcaaaaa 1200
gccgtctcag ttcggattgg gctctgcaac tcgagcccat gaagttggaa tcgctagtaa 1260
tcgcgcatca gcatggcgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca 1320
ccatgggagt tggttttacc cgaagacggt gcgctaaccg caaggaggca gccgcctcac 1380
gtaagtcagg ggggt 1395
Claims (6)
1. A mixed bacterial preparation is characterized by containing a methyl ascomycete (Methylcystis sp.) strain M9 and a Hyphomicrobium sp strain L1.
2. The mixed bacterial agent according to claim 1, wherein: in the mixed microbial inoculum, the viable bacteria content of the methylocyclosporium strain M9 and the raw silk micro-bacterium strain L1 respectively at least reaches 1.0 multiplied by 108CFU/mL。
3. The mixed bacterial agent according to claim 2, wherein: in the mixed microbial inoculum, the viable bacteria content ratio of the methylocyclosporium strain M9 to the raw silk micro-bacterium strain L1 is 1: 3-1: 15.
4. The mixed bacterial agent according to claim 1, wherein: the dosage form of the mixed microbial inoculum is powder, solution or suspension emulsion.
5. Use of the mixed bacterial agent of any one of claims 1 to 4 for removing methane and/or nitrate.
6. Use of a mixed bacterial agent according to any one of claims 1 to 4 for the removal of methane and/or nitrate in an anoxic/anaerobic environment.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4250259A (en) * | 1979-03-30 | 1981-02-10 | Exxon Research & Engineering Co. | Microbiological production of ketones from C3 -C6 secondary alcohols |
| KR101827834B1 (en) * | 2016-08-19 | 2018-02-09 | 부산대학교 산학협력단 | Novel Strain Hyphomicrobium sp. NM3 and Application Using the Same |
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