CN116925934A - Bacterial strain for degrading odor substance n-butyric acid and application thereof - Google Patents
Bacterial strain for degrading odor substance n-butyric acid and application thereof Download PDFInfo
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- CN116925934A CN116925934A CN202210336760.4A CN202210336760A CN116925934A CN 116925934 A CN116925934 A CN 116925934A CN 202210336760 A CN202210336760 A CN 202210336760A CN 116925934 A CN116925934 A CN 116925934A
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- Prior art keywords
- strain
- butyric acid
- pig manure
- manure water
- degrading
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- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 title claims abstract description 48
- 239000000126 substance Substances 0.000 title claims abstract description 18
- 230000000593 degrading effect Effects 0.000 title claims abstract description 14
- 230000001580 bacterial effect Effects 0.000 title description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 36
- 210000003608 fece Anatomy 0.000 claims abstract description 32
- 239000010871 livestock manure Substances 0.000 claims abstract description 32
- 238000004321 preservation Methods 0.000 claims abstract description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 4
- 241000222175 Diutina rugosa Species 0.000 claims abstract description 4
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 4
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 238000012258 culturing Methods 0.000 claims description 4
- 239000003205 fragrance Substances 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- 230000001877 deodorizing effect Effects 0.000 claims description 2
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 abstract description 11
- 230000015556 catabolic process Effects 0.000 abstract description 6
- 238000006731 degradation reaction Methods 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 241000894006 Bacteria Species 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 238000000424 optical density measurement Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
- B01D53/84—Biological processes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/46—Removing components of defined structure
- B01D53/72—Organic compounds not provided for in groups B01D53/48 - B01D53/70, e.g. hydrocarbons
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
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- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Health & Medical Sciences (AREA)
- Environmental & Geological Engineering (AREA)
- Analytical Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a strain for degrading an odor substance N-butyric acid and application thereof, wherein the strain number is N-BA-9, and the strain is classified and named as candida rugosa @ and the strain is a strain of candida rugosa @Candida rugose) The strain is preserved in China general microbiological culture Collection center (CGMCC) with a preservation date of 2021, 11 and 2 days and a preservation number of CGMCC No.23709. The strain N-BA-9 provided by the invention can grow by taking N-butyric acid as the only carbon source, and degrade N-butyric acid. The strain N-BA-9 is mixed with the pig manure water for culture, the growth effect of the strain N-BA-9 is good, the ozone degree M.OU is reduced to 2.604 after 24 hours of culture in a state that the ozone degree M.OU of the pig manure water is 6.233, and the degradation rate is 58.22%.
Description
Technical Field
The invention relates to a strain for degrading an odor substance n-butyric acid and application thereof, belonging to the technical field of microorganisms.
Background
With the construction of national ecology civilization, pig farm odor has become a major point of social complaints. According to research, more than 500 odor substances in a pig farm are reported, the odor components change along with the change of a cultivation mode and seasons, and n-butyric acid is one of 21 characteristic odor substances for generating odor emission of pig farm manure water, belongs to volatile fatty acids, and is difficult to eliminate. In recent years, research of breeding high-efficiency microorganism strains for degrading pig farm odorants becomes one of economic and reliable methods for degrading pig farm odorants, and screening microorganism strains for efficiently degrading odorant n-butyric acid has not been reported yet.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a strain for degrading an odor substance N-butyric acid and application thereof, wherein the strain N-BA-9 is mixed with pig manure water for culture, the growth effect of the strain N-BA-9 is good, and the ozone degree is reduced by 58.22 percent after the strain is cultured for 24 hours under the state of the ozone degree 6.233 of the pig manure water.
In order to solve the technical problems, the invention provides a strain for degrading the N-butyric acid of an odor substance, wherein the strain number is N-BA-9, and the strain is classified and named as candida rugosaCandida rugose) The strain is preserved in China general microbiological culture Collection center (CGMCC) with a preservation date of 2021, 11 and 2 days and a preservation number of CGMCC No.23709.
The invention provides application of the strain in degrading the n-butyric acid of an odor substance in a pig farm.
Further, the strain N-BA-9 is mixed with the pig manure water for culture, the strain N-BA-9 grows by taking the odor substance N-butyric acid in the pig manure water as a carbon source, and the N-butyric acid is degraded.
Further, the specific method for mixed culture of the strain N-BA-9 and the pig manure water comprises the following steps: inoculating strain N-BA-9 into fresh pig manure water with an inoculum size of 5-10%, standing and culturing at pH of 6.5-8.0 and C/N of 27-35 at 25-30deg.C for 24-48 hr, and detecting odor deodorizing rate.
Further, the inoculum size of the strain N-BA-9 was 5%.
Further, the pH was 7.2 and the C/N was 31.4.
Further, the culture temperature was 30℃and the culture time was 24 hours.
The invention has the beneficial effects that: the strain N-BA-9 provided by the invention can grow by taking the odor substance N-butyric acid in pig manure water as a carbon source, and degrade N-butyric acid. The strain N-BA-9 is mixed with the pig manure water for culture, the growth effect of the strain N-BA-9 is good, the ozone degree M.OU is reduced to 2.604 after 24 hours of culture in a state that the ozone degree M.OU of the pig manure water is 6.233, and the degradation rate is 58.22%.
Drawings
FIG. 1 shows the morphology of single colonies grown by streaking strain N-BA-9 on LB medium for 48 hours;
FIG. 2 shows the morphology of the single colony of strain N-BA-9 as observed by a microscope (10X 100);
FIG. 3 shows the detection value of ozone M.OU in pig manure water without inoculating the N-BA-9 strain;
FIG. 4 shows the detection of ozone M.OU after incubation for 24 hours with the N-BA-9 strain.
Detailed Description
The invention is further described below with reference to examples. The following examples are only for more clearly illustrating the technical aspects of the present invention, and are not intended to limit the scope of the present invention.
EXAMPLE 1 isolation, screening and identification of Strain N-BA-9
1. Isolation and screening of Strain N-BA-9
45ml of distilled water is taken in a 250ml triangular flask, a certain amount of glass beads are added, sterilization is carried out for 20min at 121 ℃, and cooling is carried out to room temperature. Weighing 5g of preserved soil sample, adding into sterile water, crushing and mixing in a constant temperature shaking table at 30deg.C and 180r/min for 1-2h to free bacteria, transferring 10ml of mud-water mixture into 90ml of basic culture medium, adding n-butyric acid into the basic culture medium, and culturing and enriching at 30deg.C and 180 r/min.
Diluting enriched and domesticated bacteria liquid with sterile water to obtain 10 -1 ~10 -6 Gradient of different concentrations, 10 -4 、10 -5 、10 -6 Three gradients of bacteria-containing water of 0.1ml are coated on a selective culture medium flat plate, then the culture is carried out in an inverted incubator at a constant temperature of 30 ℃ for 2-5 d, the further separation and purification are carried out by using a flat plate dilution separation method, the purified strains are subjected to classification numbering, and the strains are stored on a bevel at 4 ℃ for standby.
2. Identification of Strain N-BA-9
After pure culture on LB medium, single colony and bacterial form of N-BA-9 strain are observed, and FIG. 1 shows single colony form of strain N-BA-9 growing on LB medium after streak culture for 48 hr; FIG. 2 shows the morphology of the single colony of strain N-BA-9 as observed by a microscope (10X 100). Colony is small round, white, neat in edge and smooth in surface. The thallus is ellipsoidal and does not produce spores.
The results of physiological and biochemical detection show that the N-BA-9 strain is a gram-positive strain, hydrolase and catalase are positive, and methyl red, indole and oxidase are negative in reaction.
The characteristic sequence of the strain is shown in a sequence table SEQ ID NO. 1 through molecular biological identification of ITS1-5.8S-ITS 2.
EXAMPLE 2 Effect of Strain N-BA-9 on degradation of pig farm malodorous substance N-butyric acid
1. Preparation of pig manure water
Mixing 20 g fresh pig manure and 80g clean water uniformly to prepare fresh pig manure water with a solid content of about 5%, and placing the fresh pig manure water in a 37 ℃ incubator for standing fermentation for 2 days to obtain pig manure water with a ozone degree M.OU of 6.233.
2. N-BA-9 bacterium suspension for degrading odor substance N-butyric acid in pig manure water
5ml of N-BA-9 strain fermentation broth was inoculated into the above 100ml of pig manure water, the pH of the pig manure water was 7.2, the C/N was 31.4, the mixture was uniformly mixed, and after standing and culturing in an incubator at 30℃for 24 hours, the ozone degree M.OU of the pig manure water was measured, and the measurement value was 2.604, as shown in FIG. 4.
3. Calculation of degradation rate of N-BA-9 strain on odor substances in pig manure water
FIG. 3 shows the detection value of the ozone degree M.OU in the pig manure water without inoculating the strain N-BA-9, wherein the degradation rate of the odor substances in the pig manure water by the strain N-BA-9 is [ (6.233-2.604)/6.233 ] ×100% = 58.22%.
4. Description of growth conditions of bacterial strain N-BA-9 in pig manure
The growth condition of the N-BA-9 strain in the pig manure water is evaluated by microscopic observation and OD measurement, and is comprehensively evaluated by adopting 10-point scoring, wherein the comprehensive score of the growth condition of the N-BA-9 strain in the pig manure water is 6 points. The result shows that the N-BA-9 strain has higher growth speed in fecal sewage, stronger reproductive capacity and better degradation capacity on N-butyric acid.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that modifications and variations could be made by those skilled in the art without departing from the technical principles of the present invention, and such modifications and variations should also be regarded as being within the scope of the invention.
Sequence listing
<110> Nanjing agricultural university
<120> bacterial strain for degrading odor substance n-butyric acid and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 493
<212> DNA
<213> Candida rugosa (Candida rugosa)
<400> 1
ggatgctcag tacggcgagt gaagcggcaa cagctcaaat ttgaaagccc gcgggcgttg 60
taatttgcag gcggatgttt tggggcgggc gctgtctacg ttccttggaa caggacgccg 120
cagagggtga gagccccgtg cgatggcgcc tctaaccgcg taaaactccg ccgacgagtc 180
gagttgtttg ggaatgcagc tccaagtggg tggtaaattc catctaaagc taaatactgg 240
cgagagaccg atagcgaaca agtacagtga tggaaagatg aaaagcactt tgaaaagaga 300
gtgaaacagc acgtgaaatt gttgaaaggg aagggtatgc gattagcggc cagcaggagg 360
tgccttctcg tgaaaaggcc gtgcaccgtc ttcggacacc gtgcgcggag atggcgaggg 420
ggcgcctgag gtctgcgaac tcgaggttgc tggcgtaatg attgcatacc acccgtcttg 480
acccccggga cca 493
Claims (7)
1. A strain for degrading an odor substance N-butyric acid is characterized in that the strain number is N-BA-9, and is classified and named as candida rugosa @Candida rugose) The strain is preserved in China general microbiological culture Collection center (CGMCC) with a preservation date of 2021, 11 and 2 days and a preservation number of CGMCC No.23709.
2. Use of the strain of claim 1 for degrading n-butyric acid, an odorant of a pig farm.
3. The use according to claim 2, wherein the strain N-BA-9 is mixed with pig manure water, and the strain N-BA-9 grows with N-butyric acid, an odor substance in pig manure water, as a carbon source, and degrades N-butyric acid.
4. The use according to claim 3, wherein the specific method for mixed culture of the strain N-BA-9 with the pig manure water is: inoculating strain N-BA-9 into fresh pig manure water with an inoculum size of 5-10%, standing and culturing at pH of 6.5-8.0 and C/N of 27-35 at 25-30deg.C for 24-48 hr, and detecting odor deodorizing rate.
5. The use according to claim 4, wherein the inoculum size of strain N-BA-9 is 5%.
6. The use according to claim 4, wherein the pH is 7.2 and the c/N is 31.4.
7. The use according to claim 4, wherein the incubation temperature is 30℃and the incubation time is 24 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210336760.4A CN116925934B (en) | 2022-03-31 | 2022-03-31 | Bacterial strain for degrading odor substance n-butyric acid and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210336760.4A CN116925934B (en) | 2022-03-31 | 2022-03-31 | Bacterial strain for degrading odor substance n-butyric acid and application thereof |
Publications (2)
| Publication Number | Publication Date |
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| CN116925934A true CN116925934A (en) | 2023-10-24 |
| CN116925934B CN116925934B (en) | 2024-03-08 |
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| CN202210336760.4A Active CN116925934B (en) | 2022-03-31 | 2022-03-31 | Bacterial strain for degrading odor substance n-butyric acid and application thereof |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI233450B (en) * | 2001-08-31 | 2005-06-01 | Academia Sinica | Recombinant Candida rugosa lipases |
| US20060035352A1 (en) * | 2004-08-16 | 2006-02-16 | Payne Mark S | Production of 3-hydroxycarboxylic acid using nitrilase |
| CN1807641A (en) * | 2005-05-11 | 2006-07-26 | 国家海洋局第一海洋研究所 | Low temperature lipase gene sequence separated from psychrophiles and its recombinant strain for expression |
| CN103468586A (en) * | 2013-08-29 | 2013-12-25 | 北京环氧环保科技发展有限公司 | Penicillium strain and application in treatment of sewage and odor pollution thereof |
| CN106754572A (en) * | 2017-03-04 | 2017-05-31 | 江西顺泉生物科技有限公司 | One bacillus amyloliquefaciens and application thereof |
| CN112725205A (en) * | 2021-02-03 | 2021-04-30 | 淮阴工学院 | Saccharomyces strain and screening method and application thereof |
-
2022
- 2022-03-31 CN CN202210336760.4A patent/CN116925934B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI233450B (en) * | 2001-08-31 | 2005-06-01 | Academia Sinica | Recombinant Candida rugosa lipases |
| US20060035352A1 (en) * | 2004-08-16 | 2006-02-16 | Payne Mark S | Production of 3-hydroxycarboxylic acid using nitrilase |
| CN1807641A (en) * | 2005-05-11 | 2006-07-26 | 国家海洋局第一海洋研究所 | Low temperature lipase gene sequence separated from psychrophiles and its recombinant strain for expression |
| CN103468586A (en) * | 2013-08-29 | 2013-12-25 | 北京环氧环保科技发展有限公司 | Penicillium strain and application in treatment of sewage and odor pollution thereof |
| CN106754572A (en) * | 2017-03-04 | 2017-05-31 | 江西顺泉生物科技有限公司 | One bacillus amyloliquefaciens and application thereof |
| CN112725205A (en) * | 2021-02-03 | 2021-04-30 | 淮阴工学院 | Saccharomyces strain and screening method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| N. ANNAPURNA DEVI 等: ""Lipase catalyzed transesterification of ethyl butyrate synthesis in n-hexane— a kinetic study"", 《J FOOD SCI TECHNOL》, pages 1 - 7 * |
| 徐启峻 等: ""一株柱状假丝酵母产脂肪酶条件的优化研究"", 《中国油脂》, vol. 40, no. 5, pages 70 - 75 * |
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