CN114045239A - Pan-cultured paracoccus YBH-7 with dimethylacetamide degradation capability and application thereof - Google Patents
Pan-cultured paracoccus YBH-7 with dimethylacetamide degradation capability and application thereof Download PDFInfo
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Abstract
The invention discloses a Paracoccus pantoea (Paracoccus pantophus) YBH-7 with Dimethylacetamide (DMAC) degradation capability and degradation application thereof, wherein the Paracoccus pantoea YBH-7 is preserved in China center for type culture collection with the address: china, wuhan university, zip code: 430072, deposit number: CCTCC NO: m2021656, date of deposit 2021, 6 months, 01 days; the paracoccus pantotrophus YBH-7 with DMAC degradation performance provided by the invention can react to the initial concentration of 500 mg.L within 48h‑1The degradation rate of DMAC reaches 100 percent, and the discovery of the degrading bacteria is about the effect of containing DMACThe high-efficiency purification of industrial wastewater has important significance.
Description
Technical Field
The invention relates to the field of microorganisms, in particular to a pantococcus YBH-7 with dimethylacetamide degradation capability and application thereof.
Background
Dimethylacetamide (DMAC) is an important chemical raw material and a solvent with excellent performance, and is widely applied to the industries of polyurethane, acrylon, medicine, pesticide, dye, electronics and the like. DMAC has certain toxicity, can have toxic action on the environment and human beings when being discharged into the environment, and is difficult to biodegrade.
DMAC is synthetic fiber like the important solvent in acrylonitrile, polyurethane weaving and polyurethane resin synthesis industry, also can regard as C8 fraction separation styrene extraction solvent to be used simultaneously, because its extremely strong intersolubility, DMAC also receives extensive use in medicine, polymer film, the synthetic field of dyestuff coating. The DMAC wastewater mainly comes from industries such as dye, leather manufacturing, chemical synthesis and the like, and is high in concentration, strong in toxicity and difficult to degrade. The biochemical method is a method for biologically degrading pollutants in wastewater by using bacteria, and has the process characteristics of low energy consumption, high treatment efficiency, low treatment cost and the like. In the water treatment industry today, biochemical methods are the most widely used process flow and are recognized in water treatment processes in many industries. Among different wastewater treatment methods, biochemical treatment has the advantages of low cost, high treatment efficiency and no secondary pollution, so that the method is widely used and is a water treatment process with the greatest application. How to improve the efficiency of biochemical treatment is followed by research and attention of people. The biological strengthening technology, namely the biological strengthening technology, is a method for screening some dominant bacteria from sludge or soil and putting the dominant bacteria into wastewater, and can be used for specifically removing one or more pollutants in the wastewater so as to improve the biological treatment effect of the wastewater. This technology has been generated in the mid seventies of the last century and has gained considerable attention and development in the future. The main action mechanisms of bioaugmentation include direct action of highly effective degrading bacteria and co-metabolism of microorganisms. The direct action of the high-efficiency degrading bacteria is the most widely and most common action mode of the application of the microbial degradation technology, and the high-efficiency degrading strains which take target pollutants as unique carbon sources or nitrogen sources and other energy sources are thrown into wastewater after enrichment and domestication, so that the microorganisms can directly degrade the target pollutants in water to achieve the purpose of removing the pollutants. The microbial co-metabolism refers to a process that certain microbes and primary energy substances can degrade pollutants together only when the microbes exist along with the primary energy substances.
Disclosure of Invention
In order to solve the technical problem, the invention provides a paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capability and application thereof in dimethylacetamide degradation.
The specific technical scheme of the invention is as follows:
in a first aspect, the invention provides a Paracoccus pantoea with dimethylacetamide degradation capacity, which is classified and named as Paracoccus pantoea (Paracoccus pantophus) YBH-7 and has been deposited in China Center for Type Culture Collection (CCTCC) at 6.1.2021 with the addresses: wuhan university in Wuhan, China, the postal code is 430072; the preservation number is CCTCC NO. M2021656, the 16S rRNA sequence of YBH-7 is shown in SEQ ID NO. 1.
The paracoccus pantotrophus YBH-7 is derived from sludge in an aerobic tank of a certain medical wastewater treatment plant in Zhejiang, is obtained by separation and purification, and can utilize dimethylacetamide as the only carbon source for growth.
The paracoccus pantotrophus YBH-7 is characterized in that: the colony color is white, the colony is small-size single colony, no spore, opaque, the surface is smooth, the edge is neat, the form of observing this thallus under the scanning electron microscope is the bacillus.
In a second aspect, the invention provides a strain of bacteria-containing suspension taking paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capacity as an active ingredient and a preparation method of the bacteria-containing suspension.
The strain-containing suspension is prepared by performing slant culture, seed culture and fermentation on pantococcus YBH-7 with dimethylacetamide degradation capability.
The preparation method of the bacteria-containing suspension comprises the following specific steps:
(1) slant culture: inoculating paracoccus YBH-7 with dimethylacetamide degradation capability to a slant culture medium, and culturing at 30-38 deg.C for 1-3 daysObtaining inclined-plane thalli; the final concentration of the slant culture medium is as follows: k2HPO41500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, the pH value is 7.0-8.0, and the agar is 18-20 g.L-1；
(2) Seed culture: selecting colony from the slant thallus, inoculating to seed culture medium, and culturing at 30-38 deg.C for 18-24 hr to obtain seed solution; the final concentration of the seed culture medium is as follows: NaCl 10 g.L-15 g.L of yeast extract powder-1Peptone 10 g. L-1The solvent is water, and the pH value is 7.0-8.0;
(3) fermentation: inoculating the seed solution to a fermentation culture medium by an inoculation amount with a volume concentration of 1%, and culturing at 30-38 ℃ for 18-24h to obtain a fermentation culture solution, namely a bacterium-containing suspension; the final concentration of the fermentation medium is as follows: k2HPO4 1500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, and the pH value is 7.0-8.0.
In a third aspect, the invention provides an application of a bacterial suspension taking paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capability as an active ingredient and a specific method thereof in the degradation of dimethylacetamide.
The specific method comprises the following steps: inoculating the bacteria-containing suspension to a solution containing the bacteria at a final concentration of 500 mg-L-1In the liquid selective culture medium of the dimethylacetamide, the dimethylacetamide is taken as the only carbon source, and the shaking culture is carried out on a constant temperature shaking table at the temperature of 30-40 ℃ and the rotation speed of 160-300rpm, so as to obtain a culture solution; the final concentration composition of the liquid selection medium is as follows: k2HPO4 1500mg·L-1，KH2PO4500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, and the pH value is 8.0-9.0.
The invention has the beneficial effects that: the present invention providesA strain of paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capability and application of the strain in removing dimethylacetamide in wastewater, wherein the strain can react with an initial concentration of 500 mg.L within 48 hours-1The degradation rate of the DMAC reaches 100%, and the discovery of the degrading bacteria has important significance for further research on the biodegradation of the DMAC.
Drawings
FIG. 1 is a sample diagram and a scanning electron microscope diagram of strain YBH-7, wherein FIG. 1(a) is a sample diagram of strain YBH-7, and FIG. 1(b) is a scanning electron microscope diagram of strain YBH-7;
FIG. 2 is a phylogenetic tree diagram of strain YBH-7;
FIG. 3 is a comparison of strain YBH-7 with a control group for DMAC removal performance;
FIG. 4 is a graph of test results of different DMAC initial concentrations, wherein FIG. 4(a) is a graph of the change in Dimethylacetamide (DMAC) concentration at different DMAC initial concentrations, and FIG. 4(b) is a graph of the bacterial density (OD) at different DMAC initial concentrations600) A variation graph;
FIG. 5 variation of Dimethylacetamide (DMAC) concentration and bacterial density (OD) at different temperatures600) A variation graph;
FIG. 6 variation of Dimethylacetamide (DMAC) concentration and bacterial density (OD) at different pH600) A variation graph;
FIG. 7 is a graph showing the change in concentration of Dimethylacetamide (DMAC) and the change in density of bacteria (OD600) at different inoculum sizes.
Detailed Description
The invention provides a pantographs Paracoccus with dimethylacetamide degradation capability, which is classified and named as Paracoccus pantoea (Paracoccus pantophus) YBH-7, and is preserved in China Center for Type Culture Collection (CCTCC) at 6.1.2021, with the preservation number of M2021656; the 16S rRNA sequence of the YBH-7 is shown as SEQ ID NO. 1.
The invention provides a bacterium-containing suspension taking paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capability as an active ingredient, and the bacterium-containing suspension is prepared by performing slant culture, seed culture and fermentation on the paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capability.
The preparation method of the bacteria-containing suspension comprises the following specific steps:
(1) slant culture: inoculating paracoccus YBH-7 with dimethylacetamide degradation capability to a slant culture medium, and culturing at 30-38 ℃ for 1-3 days to obtain slant thalli; the final concentration of the slant culture medium is as follows: k2HPO41500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, the pH value is 7.0-8.0, and the agar is 18-20 g.L-1；
(2) Seed culture: selecting colony from the slant thallus, inoculating to seed culture medium, and culturing at 30-38 deg.C for 18-24 hr to obtain seed solution; the final concentration of the seed culture medium is as follows: NaCl 10 g.L-15 g.L of yeast extract powder-1Peptone 10 g. L-1The solvent is water, and the pH value is 7.0-8.0;
(3) fermentation: inoculating the seed solution to a fermentation culture medium by an inoculation amount with a volume concentration of 1%, and culturing at 30-38 ℃ for 18-24h to obtain a fermentation culture solution, namely a bacterium-containing suspension; the final concentration of the fermentation medium is as follows: k2HPO4 1500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, and the pH value is 7.0-8.0.
The invention provides application of a bacterial suspension taking paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capability as an active ingredient in degrading dimethylacetamide.
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto.
The final concentration composition of the liquid selection medium used in the present invention is: k2HPO4 1500mg·L-1，KH2PO4500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water.
Example 1: isolation, purification and identification of Paracoccus pantoea (Paracoccus pantophus) YBH-7
1. Isolation and purification of Paracoccus pantoea (Paracoccus pantophus) YBH-7
Paracoccus pantoea (Paracoccus pantophus) YBH-7 is screened from sludge in an aerobic tank of a certain medical wastewater treatment plant in Taizhou, Zhejiang, and comprises the following specific steps:
(1) sampling: sampling sludge in an aerobic tank of a certain medical wastewater treatment plant in Zhejiang at multiple points respectively to be used as a raw material for screening Paracoccus pantoea (Paracoccus pantophus) YBH-7 with dimethylacetamide degradation capacity;
(2) strain separation: taking a proper amount of activated sludge in an aerobic pool, standing for 2 hours, taking 10mL of supernatant, inoculating the supernatant into a 250mL culture bottle containing 100mL of enrichment culture solution, carrying out shake culture on a constant-temperature shaking table at 30 ℃ and 160rpm for 24 hours, and repeating for 3 times, wherein the final concentration components of the enrichment culture solution are as follows: tryptone 2.5 g.L-11.25 g.L yeast extract -1500 mg.L of agar powder-1，K2HPO4500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1(ii) a Diluting the bacterial liquid with sterile water 10-2、10-3、10-4、10-5、10-6、10-7Doubling; and (3) separating and purifying the obtained bacterial liquid by using a liquid selective culture medium through multiple flat plate streaking to obtain a single bacterial colony, and marking the single bacterial colony as a bacterial strain YBH-7.
2. Identification of Strain YBH-7
a. Physiological and biochemical characteristics of strain YBH-7
Morphological observation and physiological and biochemical identification are carried out on the obtained strain YBH-7, and the result shows that the strain YBH-7 can be cultured in a common bacterium LB culture medium and has the final concentration of 500 mg.L-1The liquid selective culture medium of the dimethylacetamide grows, the colony color is white, the colony is a small single colony, has no spore, is opaque, has a smooth surface and is neat in edge, the strain YBH-7 is shown in figure 1(a), the shape of the thallus observed under a scanning electron microscope is bacillus shown in figure 1(b), the optimal pH value for growth is 8.0, and the optimal temperature is 35 ℃.
b. 16S rRNA sequence analysis of strain YBH-7
The strain YBH-7 is determined to be Paracoccus pantophus through 16S rRNA sequence analysis and physiological and biochemical experiment identification. The sequencing result is as follows:
cgcacccttcggggtgagcggcggacgggtgagtaacgcgtgggaatatgccctttggtacggaatagtcctgggaaactgggggtaataccgtatgcgcccttcgggggaaagatttatcgccaaaggattagcccgcgttggattaggtagttggtggggtaatggcctaccaagccgacgatccatagctggtttgagaggatgatcagccacactgggactgagacacggcccagactcctacgggaggcagcagtggggaatcttagacaatgggggcaaccctgatctagccatgccgcgtgagtgatgaaggccctagggttgtaaagctctttcagctgggaagataatgacggtaccagcagaagaagccccggctaactccgtgccagcagccgcggtaatacggagggggctagcgttgttcggaattactgggcgtaaagcgcacgtaggcggaccggaaagttgggggtgaaatcccggggctcaaccccggaactgccttcaaaactatcggtctggagttcgagagaggtgagtggaattccgagtgtagaggtgaaattcgtagatattcggaggaacaccagtggcgaaggcggctcactggctcgatactgacgctgaggtgcgaaagcgtggggagcaaacaggattagataccctggtagtccacgccgtaaacgatgaatgccagtcgtcgggcagcatgctgttcggtgacacacctaacggattaagcattccgcctgggggagtacggtcgcaagattaaaactcaaaggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgcagaaccttaccaacccttgacatcccaggaccggcccggagacgggtctttcacttcggtgacctggagacaggtgctgcatggctgtcgtcagctcgtgtcgtgagatgttcggttaagtccggcaacgagcgcaacccacactcttagttgccagcatttggttgggcactctaagagaactgccgatgataagtcggaggaaggtgtggatgacgtcaagtcctcatggcccttacgggttgggctacacacgtgctacaatggtggtgacagtgggttaatccccaaaagccatctcagttcggattggggtctgcaactcgaccccatgaagttggaatcgctagtaatcgcggaacagcatgccgcggtgaatacgttcccgggccttgtacacaccgcccgtcacaccatgggagttgggtctacccgacg。
uploading the 16S rDNA sequence of YBH-7 to Genbank to obtain the accession number MZ144121 of Genbank, and performing homology comparison with the gene sequence in Genbank, wherein FIG. 2 is the phylogenetic tree diagram of the strain. In order to further determine the reliability of the identification result, the strain YBH-7 is finally determined to belong to Paracoccus pantophus through physiological and biochemical experiments, and therefore, the strain is named as Paracoccus pantophus (Paracoccus pantophus) YBH-7.
EXAMPLE 2 preparation of a suspension containing As active ingredient Paracoccus pantotrophus YBH-7 with dimethylacetamide-degrading ability
The preparation process of the bacteria-containing suspension comprises the following steps:
(1) slant culture: inoculating paracoccus YBH-7 with dimethylacetamide degradation capability to a slant culture medium, and culturing for 3 days at 30 ℃ to obtain slant thalli; the final concentration of the slant culture medium is as follows: k2HPO4 1500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, the pH value is 7.0, and the agar is 18 g.L-1；
(2) Seed culture: selecting a bacterial colony from the inclined-plane thallus, inoculating the bacterial colony to a seed culture medium, and culturing for 18h at 30 ℃ to obtain a seed solution; the final concentration of the seed culture medium is as follows: NaCl 10 g.L-15 g.L of yeast extract powder-1Peptone 10 g. L-1The solvent is water, and the pH value is 7.0;
(3) fermentation: inoculating the seed solution to a fermentation culture medium by an inoculation amount with the volume concentration of 1%, and culturing at 30 ℃ for 24h to obtain a fermentation culture solution, namely a bacterium-containing suspension; the final concentration of the fermentation medium is as follows: k2HPO4 1500mg·L-1，KH2PO4500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, and the pH value is 7.0.
Example 3: detection of degradation performance of Paracoccus pantoea (Paracoccus pantophus) YBH-7 on dimethylacetamide and degradation condition screening
1. Detection of degradation performance of Paracoccus pantoea (Paracoccus pantophus) YBH-7 on Dimethylacetamide (DMAC)
The bacterial suspension prepared in example 2 and containing paracoccus pantoea YBH-7 with dimethylacetamide degradation capability as active component was inoculated to a final concentration of 500 mg.L-1In the liquid selection culture medium of dimethylacetamide, dimethylacetamide is used as a unique carbon source, the initial pH value of the culture medium is 8.0, and 100m is takenPlacing the L into a 250mL triangular flask, and carrying out shake culture on a constant temperature shaking table at 30 ℃ and 160rpm to obtain a culture solution (used as an experimental group);
at the same time, 100mL of the solution is filled to contain the final concentration of 500 mg.L-1A250 mL Erlenmeyer flask of the culture medium was selected as a liquid of dimethylacetamide so that the initial pH of the culture medium was 8.0, and the culture was performed with shaking on a constant temperature shaker at 30 ℃ and 160rpm as a blank control.
During the culture, the culture solutions of the experimental group and the blank control group were extracted at intervals of 8 hours, and the concentration of dimethylacetamide was measured and a DMAC degradation rate graph was plotted, as shown in fig. 3. The detection method comprises the following steps:
the detection of the concentration of dimethylacetamide adopts High Performance Liquid Chromatography (HPLC): treating the sample, taking 1ml of culture solution every 8 hours, centrifuging the culture solution at 10000rpm for 5min to remove microorganisms to obtain a supernatant, and then carrying out HPLC quantitative analysis to obtain the concentration of the dimethylacetamide.
As can be seen from FIG. 3, the Paracoccus pantoea (Paracoccus pantoeus) YBH-7 has a good degradation effect on Dimethylacetamide (DMAC), and the initial concentration is 500 mg.L in 48h-1The degradation rate of the dimethylacetamide reaches 100 percent.
2. Detection of influence of initial concentration of Dimethylacetamide (DMAC) on degradation of dimethylacetamide by Paracoccus pantoea (Paracoccus pantoeus) YBH-7
The bacterial suspensions prepared in example 2 and containing paracoccus pantoea YBH-7 with dimethylacetamide degradation capability as active ingredients were inoculated to final concentrations of dimethylacetamide (1 g. L)-1、5g·L-1、10g·L-1、15g·L-1、20g·L-1) In the liquid selective medium (2), dimethylacetamide is used as a sole carbon source, the initial pH value of the medium is 8.0, 100mL of the medium is respectively placed in 5 triangular flasks of 250mL, the medium is subjected to shaking culture on a constant temperature shaking bed of 160rpm at 30 ℃, and the concentration of dimethylacetamide and the bacterial density (OD) are measured at intervals of 24h600) The results are shown in FIG. 4. Bacterial density (OD)600) The detection method comprises the following steps: measuring the reaction solution with Shimadzu UV2401 UV-visible spectrophotometer at a wavelength of 600nmAbsorbance.
FIG. 4(a) is a graph showing the change in Dimethylacetamide (DMAC) concentration at different initial DMAC concentrations, and FIG. 4(b) is a graph showing the bacterial density (OD) at different initial DMAC concentrations600) And (5) a variation graph.
As shown in FIG. 4, the Paracoccus pantoea (Paracoccus pantoea) YBH-7 has a good degradation effect on Dimethylacetamide (DMAC), the DMAC removal efficiency is reduced along with the increase of the initial concentration of DMAC, and the bacterial density (OD) of each group is the same600) With increasing concentration, the initial concentration is 1 g.L-1The degradation rate reaches 100 percent at 120h, and the initial concentration is 5 g.L-1The degradation rate reaches 50% at 120 h.
In conclusion, Paracoccus pantotrophus (Paracoccus communis) YBH-7 has a good degradation effect on Dimethylacetamide (DMAC), has strong degradation capability on high-concentration dimethylacetamide wastewater, and can basically reduce dimethylacetamide to a dischargeable standard in a short time.
3. Optimum temperature condition screening of Paracoccus pantoea (Paracoccus pantophus) YBH-7 on Dimethylacetamide (DMAC) degradation
The bacterial suspensions prepared in example 2 and containing paracoccus pantoea YBH-7 with dimethylacetamide degradation capability as active ingredients were respectively inoculated to a final concentration of dimethylacetamide of 500 mg.L-1In the liquid selective culture medium, the initial pH value of the culture medium is 8.0, 100mL of the culture medium is respectively placed in 5 triangular flasks of 250mL, and the triangular flasks are respectively placed on a constant temperature shaking table of 25 ℃, 30 ℃, 35 ℃, 40 ℃ or 45 ℃ and 160rpm for shaking culture; the concentration and bacterial density (OD) of dimethylacetamide was measured after 24h of shaking culture600) The results are shown in FIG. 5.
The results are shown in FIG. 5: the strain YBH-7 has good tolerance to 30 ℃, 35 ℃ and 40 ℃; the optimal temperature is 35 ℃, and the DMAC degradation rate of the strain YBH-7 reaches 33% in 24 h; the optimum growth and propagation temperature of the strain YBH-7 under the experimental condition is 30-40 ℃, and the strain can better exert degradation performance within the temperature range.
4. Optimum pH value condition screening of Paracoccus pantoea (Paracoccus pantophus) YBH-7 on Dimethylacetamide (DMAC) degradation
The bacterial suspensions prepared in example 2 and containing paracoccus pantoea YBH-7 with dimethylacetamide degradation capability as active ingredients were respectively inoculated to a final concentration of dimethylacetamide of 500 mg.L-1In the liquid selective culture medium of (1), 100mL of the culture medium is respectively placed in 5 triangular flasks of 250mL, the pH values of the culture medium are respectively adjusted to 5.0, 6.0, 7.0, 8.0 or 9.0, and shaking culture is carried out on a constant temperature shaking bed of 160rpm at 30 ℃; the concentration and bacterial density (OD) of dimethylacetamide was measured after 24h of shaking culture600) The measured results are shown in fig. 6;
the results are shown in FIG. 6: the strain YBH-7 has good tolerance to pH values of 7.0, 8.0 and 9.0; the pH value of 8.0 is taken as the best, and the DMAC degradation rate of the strain YBH-7 reaches 40% in 24 h; the optimum growth and reproduction pH value of the strain YBH-7 under the experimental condition is 7.0-9.0, and the strain can better exert degradation performance within the pH value range.
5. Screening of optimal inoculum size of Paracoccus pantoea (Paracoccus pantophus) YBH-7 for Dimethylacetamide (DMAC) degradation
Inoculating the seed solution in the step (3) of the example 2 into a fermentation medium by the inoculation amount of 1%, 2%, 3%, 4% and 5% in volume concentration respectively, and preparing the bacterial-containing suspension with the inoculation amount of 1%, 2%, 3%, 4% and 5% and the paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capability as an active ingredient without changing other steps of the example 2.
Respectively inoculating 1%, 2%, 3%, 4%, 5% of strain-containing suspension containing Paracoccus pantropic YBH-7 with dimethylacetamide degradation capability as active component to dimethylacetamide with final concentration of 500 mg.L-1In the liquid selective medium of (1), 100mL of each of the culture media was placed in 5 250mL triangular flasks at an initial pH of 8.0, and shake-cultured on a constant temperature shaker at 30 ℃ and 160 rpm; the concentration and bacterial density (OD) of dimethylacetamide was measured after 24h of shaking culture600) The measured results are shown in fig. 7;
the results are shown in FIG. 7: the DMAC degradation rate measured in 24h is increased along with the increase of the inoculation amount; when the inoculation amount is 5%, the DMAC degradation rate of the strain YBH-7 reaches 84% in 24 h.
Sequence listing
<110> Zhejiang tree college (Zhejiang tree university)
<120> paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capacity and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1030
<212> RNA
<213> Paracoccus pan-cultured (Paracoccus pantrophus)
<400> 1
cgcaccccgg gggagcggcg gacggggaga acgcggggaa agcccggacg gaaagccggg 60
aaacggggga aaccgagcgc cccgggggaa agaacgccaa aggaagcccg cgggaaggag 120
ggggggaagg ccaccaagcc gacgaccaag cgggagagga gacagccaca cgggacgaga 180
cacggcccag acccacggga ggcagcaggg ggaacagaca agggggcaac ccgacagcca 240
gccgcggagg agaaggccca ggggaaagcc cagcgggaag aaagacggac cagcagaaga 300
agccccggca acccggccag cagccgcgga aacggagggg gcagcggcgg aaacgggcga 360
aagcgcacga ggcggaccgg aaaggggggg aaacccgggg ccaaccccgg aacgcccaaa 420
acacggcgga gcgagagagg gagggaaccg aggagaggga aacgagaacg gaggaacacc 480
agggcgaagg cggccacggc cgaacgacgc gagggcgaaa gcgggggagc aaacaggaag 540
aacccggagc cacgccgaaa cgagaagcca gcgcgggcag cagcgcggga cacaccaacg 600
gaaagcaccg ccgggggaga cggcgcaaga aaaaccaaag gaagacgggg gcccgcacaa 660
gcggggagca gggaacgaag caacgcgcag aaccaccaac ccgacaccca ggaccggccc 720
ggagacgggc caccgggacc ggagacaggg cgcaggcgcg cagccggcgg agagcggaag 780
ccggcaacga gcgcaaccca caccaggcca gcagggggca ccaagagaac gccgagaaag 840
cggaggaagg gggagacgca agcccaggcc cacggggggc acacacggca caagggggac 900
aggggaaccc caaaagccac cagcggaggg gcgcaaccga ccccagaagg gaacgcagaa 960
cgcggaacag cagccgcggg aaacgcccgg gccgacacac cgcccgcaca ccagggaggg 1020
gcacccgacg 1030
Claims (5)
1. A strain of Paracoccus pantotrophus with dimethylacetamide degradation capability is characterized in that microorganisms are classified and named as Paracoccus pantotrophus (Paracoccus pantoprazus) YBH-7, and are preserved in China Center for Type Culture Collection (CCTCC) at 6 months and 1 days in 2021, wherein the preservation number is M2021656; the 16S rRNA sequence of the YBH-7 is shown as SEQ ID NO. 1.
2. An bacteria-containing suspension with paracoccus pantotrophus as an active ingredient in claim 1, wherein the bacteria-containing suspension is prepared by performing slant culture, seed culture and fermentation on paracoccus pantotrophus YBH-7 with dimethylacetamide degradation capacity.
3. A method for preparing a suspension containing bacteria according to claim 2, which comprises the following steps:
(1) slant culture: inoculating paracoccus YBH-7 with dimethylacetamide degradation capability to a slant culture medium, and culturing at 30-38 ℃ for 1-3 days to obtain slant thalli; the final concentration of the slant culture medium is as follows: k2HPO41500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, the pH value is 7.0-8.0, and the agar is 18-20 g.L-1；
(2) Seed culture: selecting colony from the slant thallus, inoculating to seed culture medium, and culturing at 30-38 deg.C for 18-24 hr to obtain seed solution; the final concentration of the seed culture medium is as follows: NaCl 10 g.L-15 g.L of yeast extract powder-1Peptone 10 g. L-1The solvent is water, the pH value is 7.0-8.0；
(3) Fermentation: inoculating the seed solution to a fermentation culture medium by an inoculation amount with a volume concentration of 1%, and culturing at 30-38 ℃ for 18-24h to obtain a fermentation culture solution, namely a bacterium-containing suspension; the final concentration of the fermentation medium is as follows: k2HPO4 1500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, and the pH value is 7.0-8.0.
4. Use of the bacterial-containing suspension of claim 2 for the degradation of dimethylacetamide.
5. The use according to claim 4, characterized in that it is in particular: inoculating the bacteria-containing suspension to a solution containing the bacteria at a final concentration of 500 mg-L-1In the liquid selective culture medium of the dimethylacetamide, the dimethylacetamide is taken as the only carbon source, and the shaking culture is carried out on a constant temperature shaking table at the temperature of 30-40 ℃ and the rotation speed of 160-300rpm, so as to obtain a culture solution; the final concentration composition of the liquid selection medium is as follows: k2HPO4 1500mg·L-1，KH2PO4 500mg·L-1，NaCl 1000mg·L-1，DMAC 500mg·L-1，MgSO4·7H2O 200mg·L-1The solvent is water, and the pH value is 7.0-9.0.
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| CN117126781A (en) * | 2023-09-01 | 2023-11-28 | 贵州省材料产业技术研究院 | Arthrobacter protoglass for degrading dimethylacetamide and application thereof |
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