CN106754462B - Sphingobacterium desulfurates and application thereof - Google Patents
Sphingobacterium desulfurates and application thereof Download PDFInfo
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- 241001136275 Sphingobacterium Species 0.000 title claims abstract description 20
- 238000006477 desulfuration reaction Methods 0.000 claims abstract description 36
- 230000023556 desulfurization Effects 0.000 claims abstract description 36
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims abstract description 24
- WWUZIQQURGPMPG-UHFFFAOYSA-N (-)-D-erythro-Sphingosine Natural products CCCCCCCCCCCCCC=CC(O)C(N)CO WWUZIQQURGPMPG-UHFFFAOYSA-N 0.000 claims abstract description 17
- WWUZIQQURGPMPG-KRWOKUGFSA-N sphingosine Chemical compound CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](N)CO WWUZIQQURGPMPG-KRWOKUGFSA-N 0.000 claims abstract description 17
- 241000894006 Bacteria Species 0.000 claims abstract description 16
- 238000000034 method Methods 0.000 claims abstract description 10
- 230000008569 process Effects 0.000 claims abstract description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- 230000001580 bacterial effect Effects 0.000 claims description 12
- 229910052799 carbon Inorganic materials 0.000 claims description 12
- 230000001651 autotrophic effect Effects 0.000 claims description 10
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 8
- 238000004321 preservation Methods 0.000 claims description 7
- 235000016709 nutrition Nutrition 0.000 claims description 6
- 230000035764 nutrition Effects 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- 235000015278 beef Nutrition 0.000 claims description 4
- 239000001569 carbon dioxide Substances 0.000 claims description 4
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 150000004763 sulfides Chemical class 0.000 claims description 4
- 210000003495 flagella Anatomy 0.000 claims description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims description 2
- 239000001301 oxygen Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 150000003467 sulfuric acid derivatives Chemical class 0.000 claims description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 claims 1
- 150000003464 sulfur compounds Chemical class 0.000 claims 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 abstract description 28
- 241000193830 Bacillus <bacterium> Species 0.000 abstract description 13
- 238000006243 chemical reaction Methods 0.000 abstract description 9
- 229910052717 sulfur Inorganic materials 0.000 abstract description 8
- 239000011593 sulfur Substances 0.000 abstract description 8
- 239000002351 wastewater Substances 0.000 abstract description 6
- 230000007613 environmental effect Effects 0.000 abstract description 4
- 238000000926 separation method Methods 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract 1
- 150000003568 thioethers Chemical class 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 description 21
- 229910052979 sodium sulfide Inorganic materials 0.000 description 11
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 description 11
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 7
- 235000019345 sodium thiosulphate Nutrition 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 244000005700 microbiome Species 0.000 description 6
- 238000012216 screening Methods 0.000 description 6
- 241001052560 Thallis Species 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000002245 particle Substances 0.000 description 5
- 239000007836 KH2PO4 Substances 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 241000605268 Thiobacillus thioparus Species 0.000 description 4
- 238000005273 aeration Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L magnesium chloride Substances [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
- 229910001629 magnesium chloride Inorganic materials 0.000 description 4
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 239000011573 trace mineral Substances 0.000 description 4
- 235000013619 trace mineral Nutrition 0.000 description 4
- 239000011782 vitamin Substances 0.000 description 4
- 229940088594 vitamin Drugs 0.000 description 4
- 229930003231 vitamin Natural products 0.000 description 4
- 235000013343 vitamin Nutrition 0.000 description 4
- 150000003722 vitamin derivatives Chemical class 0.000 description 4
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 3
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 3
- 238000004626 scanning electron microscopy Methods 0.000 description 3
- 239000010865 sewage Substances 0.000 description 3
- 239000010802 sludge Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 241000685602 Sphingobacterium sp. Species 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000736131 Sphingomonas Species 0.000 description 1
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- 238000004065 wastewater treatment Methods 0.000 description 1
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- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C10L3/10—Working-up natural gas or synthetic natural gas
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- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
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Abstract
The invention relates to a new sphingosine bacterium desulfurated by separation and purification and application thereof in biological desulfurization, belonging to the technical field of biology. The invention discloses a newly screened Sphingobacterium sp DS-4 and application thereof in removing sulfur in sulfides, belonging to the technical field of biology and environmental protection. The Sphingobacterium strain of the invention has a 16SrDNA sequence shown in SEQ ID No. 1. The sphingosine bacillus strain disclosed by the invention can well grow in wastewater with high sulfide content, can tolerate higher sulfide concentration, has high sulfide removal efficiency in a desulfurization process, can achieve a 5-hour removal rate of 99.13%, can convert sulfide into recoverable elemental sulfur in the removal process, has a sulfur conversion rate of 77.9%, and has higher environmental protection value and economic benefit.
Description
Technical Field
The invention relates to a sphingosine bacillus, in particular to a newly screened desulfurated sphingosine bacillus and application thereof in biological desulfurization, belonging to the technical field of biology and environmental protection.
Background
The release of hydrogen sulfide is a main problem associated with biogas and wastewater treatment. Namely, the biogas and the hydrogen sulfide in the wastewater of the sewage treatment plant are directly or indirectly discharged into the air, which not only causes pollution to the atmospheric environment, but also threatens the health of people. Due to the problems of corrosiveness, toxicity, stink and high oxygen consumption of the sulfide, the waste gas and the waste water discharged into the environment in industrial production must enable the sulfide to reach relevant industrial standards to be discharged. Currently, commonly used desulfurization means include physical desulfurization, chemical desulfurization, and biological desulfurization.
The traditional desulfurization process usually uses a chemical method to remove sulfide, but the operation cost is higher because the desulfurizing agent needs to be continuously consumed. The biological desulfurization technology mainly relies on microorganisms capable of self-propagating to catalyze desulfurization, the desulfurizer can be self-regenerated, the operation cost is low, and the application prospect is wide.
In view of the above phenomena, in the 50 s of the 20 th century, research on desulfurization using microorganisms has been started, and biological desulfurization has been one of the commonly used methods for removing sulfides. In the prior art of biological desulfurization, autotrophic sulfur-oxidizing bacteria are often used for removing sulfides, but some problems also exist, such as too slow growth speed of bacterial strains, difficulty in obtaining high-density bacteria, low desulfurization efficiency and increased cost.
For example, an alkali-resistant thiobacillus thioparus and an application (application number: 201110149623.1) thereof in biological desulfurization have strong capability of removing hydrogen sulfide under certain conditions, but have the defects that the nutrition type of the thiobacillus thioparus is strictly autotrophic, high-density thalli are not easy to obtain quickly in practical engineering application, the growth speed of the thiobacillus thioparus is slow, and the thiobacillus thioparus needs to be cultured for 48 hours.
In conclusion, some high-efficiency desulfurization strains which grow rapidly and can rapidly remove inorganic sulfur are urgently needed in the biological desulfurization industry at present.
Disclosure of Invention
The invention aims to provide an efficient desulfurization strain and application thereof in biological desulfurization of sulfide-containing wastewater, natural gas or methane, aiming at the problems and defects of slow growth, difficult acquisition of high-density thalli and slow desulfurization rate of the existing desulfurization microorganisms.
In order to achieve the purpose, the invention firstly provides a sulfur oxidizing bacterium, namely sphingosine bacillus (Sphingobacterium sp.) DS-4, which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms at 2016, 16.06.16.7 (address: Beijing city, West Lu No.1 Hospital No. 3 of the rising district of the republic of China, institute of microbiology, postal code: 100101), wherein the preservation number is as follows: CGMCC No. 12632.
The sphingosine bacillus DS-4 provided by the invention is obtained by screening sludge in an aerobic pool of a second sewage treatment plant in metropolis, and the screening method comprises the following steps: adopting a sodium thiosulfate culture medium for enrichment and domestication for 7 days, repeating for 3-4 times, adopting a sodium sulfide culture medium for plate separation, generating bacterial colonies after 3-4 days, selecting a single bacterial colony to be inoculated into the sodium sulfide culture medium for culture, repeating for more than 4 times to obtain pure bacteria, and having the physiological characteristics that: the bacterium is in the shape of a short rod, 1.2-2 mu m long and 0.8-0.1.2 mu m wide, is gram-negative and has no flagella; the growth pH is 5.0-9.0, and the optimum pH is 7.0; the growth temperature is 20-35 ℃, and the optimal temperature is 30 ℃; the bacterial colony is oval, the surface is concave, and the color is milky white; the nutrition type of the strain is facultative, and the strain can utilize organic carbon sources such as beef extract, peptone and yeast powder to perform heterotrophic rapid growth and can also utilize carbon dioxide as a unique carbon source to perform autotrophic growth; when the self-curing process is carried out by taking carbon dioxide as the only carbon source, the bacteria can utilize oxygen to oxidize sulfides into elemental sulfur or sulfates.
2HS-+O2→2S+2OH-
HS-+2O2→SO4 2-+H+
Secondly, the invention also provides the application of the strain CGMCC No.12632, and the desulfurization effect verification shows that the strain realizes 99.13 percent of sulfide removal and 77.9 percent of elemental sulfur conversion rate within 5 hours under the autotrophic condition. The strain is used for biological desulfurization.
The invention also provides a culture medium used in the processes of strain screening and culture, wherein the formula of the culture medium is obtained by optimization in the domestication and enrichment process, and the formula is as follows: (1) sodium sulfide culture medium for strain screening and desulfurization effect experiment, and its preparation methodThe formula is as follows: na (Na)2S.9H2O 10.0g，KH2PO41.0g，K2HPO41.0g，NH4Cl 0.2g，MgCl20.2g，NaHCO31.0g, 1ml of trace element solution and 1ml of vitamin solution, and adjusting the pH to be neutral by using 1mol/L HCl solution. (2) The sodium thiosulfate culture medium is used for strain enrichment, and the formula of the culture medium is as follows: na (Na)2S2O3.5H2O 10.0g，KH2PO41.0g，K2HPO41.0g，NH4Cl0.2g，MgCl20.2g，NaHCO31.0g, 1ml of trace element solution and 1ml of vitamin solution, and adjusting the pH to be neutral by using 1mol/L NaOH or HCl solution. (3) An organic carbon source heterotrophic culture medium is used for expanding culture and verifying the nutrition type of the strain: 5.0g/L beef extract, 10.0g/L peptone, 5.0g/L sodium chloride and pH 7.0-7.2.
The invention provides the 16SrDNA identification result of the strain. By 16SrDNA identification and BLAST comparison, the homology of DS-4 and Sphingobacterium reaches 99 percent, so that the strain is identified as Sphingobacterium DS-4. The 16SrDNA gene sequence of the sphingosine bacillus provided by the invention is shown as SEQ ID No. 1:
GAAGGATGGCGTCTGTCTACAATGCAGTCGAGCGCACCCTTCGGGGTGCTCGGTGTCCGGGTGAATAGTGCGTGACGGTAGGCCCTTTGGTGCGCAATCTACCCGTATCACGGGGATAGCCCCAGTATGCGCCCTTCGGGGCAAAGATTTATCTCCCCCGGCATCCCCGCGTTGGATTAAGTTATTGGAGGGGTATGGGCTCACGTGACCTAAGATCCTTGGTGGTTTGAGAGGATGATCAGCGAACATGTCACTGAGATCTGACCAGACTCCTACCACACGGGTACTGGGACAATCTTAAAACTCCTACGCAACCCTGATCTAACGATGCCTGGTGAATGATGACAGCCCTGAACCTGCCATGCTCTTTGCACGATGACTGCCCTATGGGTTGTAAACTGCTTTTGTTAGGCTAACTCCCCGTCTACATGTACGTGGCTGAATGTACCGCTAGCGTTGTGCTCAATTACTGGCCGGCCAGCGCACGTAGTAATACCGAAGATCCGAGCGTTATCCGGATTTATTGGGTTTAAGAGGTGCCTACGCGGTACTTTAAGTCGAGGGTGAGAGACTGCGTGTTATTGTCGTGTGCCTTTGATACTGAAGTGCTTGAATGCAATACAAGACGGCGGAGTGAGACACTGACTCGATACATGCATAGAAGTGTCTCAGAATGCCGATTGCGAAAGGATCTGTCTACTGCGTTACTGACCCCTGATGCAAAAAATGCTGGTCATCGAACAGCATGATGTACCCTTGACAGACCCTACCCCTATAAAGATGACAACTCCGATGATTGCACCCTACCCGATCACATCCCAAACGAAATCGATAAGTTTCTCCATCTTGGCGAGGGCGCTTCGTGCGTTTTGTATTTA
the relevant preservation information related to the invention is as follows:
the preservation unit: china general microbiological culture Collection center; address: western road No.1 institute of north chen, institute of microbiology, academy of sciences of china, tokyo, japan; the preservation date is as follows: 2016, month 06, 16; the preservation number is: CGMCC No. 12632; is named as: sphingobacterium sp DS-4.
The invention has the advantages that:
(1) the nutrition type of the sphingosine bacillus related by the invention is facultative, the rapid propagation of the thallus is realized under the heterotrophic condition, and the high-efficiency desulfurization is realized under the autotrophic condition. The strain is subjected to amplification culture by using an organic carbon source heterotrophic culture medium to obtain high-density thalli, and the high-density thalli is used for realizing the quick start of the biological desulfurization engineering.
(2) The desulfurization rate of the sphingosine bacillus related by the invention is higher, the removal rate of sulfide can reach 99.13% in 5h under the autotrophic condition, and the conversion rate of elemental sulfur can reach 77.9%;
(3) the sphingosine bacillus can tolerate sulfide with higher concentration and sulfur (S) tolerant under autotrophic condition2-) The concentration is up to 1098.75 mg/L;
(4) the microbial strain is obtained by screening from a selection environment stressed by high sulfide for a long time and does not cause harm to the surrounding environment and ecological balance.
Drawings
FIG. 1 cellular morphology of Sphingobacterium DS-4 of the present invention under Scanning Electron Microscopy (SEM).
FIG. 2 is a graph showing the growth curve of Bacillus sphingomonas DS-4 according to the present invention at an initial pH of 7.0 and a temperature of 30 ℃ wherein: (a) a growth curve of Sphingobacterium DS-4 under autotrophic conditions, (b) a growth curve of Sphingobacterium DS-4 under heterotrophic conditions.
FIG. 3 shows the variation of various factors in the course of desulfurization effect experiment of Sphingobacterium DS-4 of the present invention, wherein: (a) a change in sulfide removal effect, (b) a change in elemental sulfur concentration.
FIG. 4 morphology of biosulfur particles formed by desulfurization of Sphingobacterium DS-4 of the present invention under Scanning Electron Microscopy (SEM).
Detailed Description
The present invention will be described in detail with reference to examples. The embodiments are provided to facilitate a better understanding of the invention and are not intended to limit the invention.
The medium formulations referred to in the examples were:
(1) the sodium sulfide culture medium comprises the following components in percentage by weight: na (Na)2S.9H2O 10.0g，KH2PO41.0g，K2HPO41.0g，NH4Cl0.2g，MgCl20.2g，NaHCO31.0g, 1ml of trace element solution and 1ml of vitamin solution, and adjusting the pH to be neutral by using 1mol/L HCl solution.
(2) The sodium thiosulfate culture medium comprises the following components in percentage by weight: na (Na)2S2O3.5H2O 10.0g，KH2PO41.0g，K2HPO41.0g，NH4Cl 0.2g，MgCl20.2g，NaHCO31.0g, 1ml of trace element solution and 1ml of vitamin solution, and adjusting the pH to be neutral by using 1mol/L NaOH or HCl solution.
(3) The organic carbon source heterotrophic culture medium comprises the following components in percentage by weight: 5.0g/L beef extract, 10.0g/L peptone, 5.0g/L sodium chloride and pH 7.0-7.2.
Example 1: strain screening
Taking a proper amount of sludge in an aerobic aeration tank of a second sewage treatment plant in metropolis, adding the sludge into a reactor containing a 5L sodium thiosulfate culture medium according to the inoculation amount of 20%, carrying out enrichment culture for 7 days under the conditions of pH 7.0, aeration 0.5L/min and 30 ℃, repeating for 3-4 times, carrying out plate separation by adopting a sodium sulfide culture medium, generating bacterial colonies after 3-4 days, selecting a single bacterial colony, inoculating the single bacterial colony into the sodium sulfide culture medium for culture, and repeating for more than 4 times to obtain pure bacteria.
Example 2: identification of strains
Inoculating the strain into a sodium thiosulfate culture medium according to the same inoculation amount, and culturing in shaking tables (20 ℃, 25 ℃, 30 ℃, 35 ℃ and 40 ℃) at different temperatures for 30h to determine the optimal growth temperature of the strain to be 30 ℃; culturing at the optimum growth temperature for 30h with initial pH of 5.0, 6.0, 7.0, 8.0, and 9.0, respectively, and determining the optimum growth pH to be 7.0.
The separated and purified strain was observed under a Scanning Electron Microscope (SEM), and the results are shown in FIG. 1; the strain is in the shape of a short rod, 1.2-2 mu m long and 0.8-0.1.2 mu m wide and has no flagella; gram identifies the bacterium as negative.
A bacterial whole genome rapid extraction kit is adopted to extract the whole genome of a pure strain, PCR is carried out through 16SrDNA universal primers 27F and 1492R, then sequencing analysis is carried out, the sequencing result is compared through BLAST, the strain is identified to be sphingosine bacillus, and the strain is named DS-4, namely the preservation strain sphingosine bacillus CGMCC No.12632(Sphingobacterium sp.DS-4).
The growth curve study of Sphingobacterium DS-4 was carried out, and sodium thiosulfate medium and organic carbon source heterotrophic medium were selected for autotrophic culture and heterotrophic culture, respectively, and the results are shown in the following FIGS. 2(a), (b): (a) the Sphingobacterium DS-4 can grow in a sodium thiosulfate culture medium, and the OD value of a bacterial liquid reaches 0.21 after the bacterial liquid is cultured for 30 hours; (b) the Sphingobacterium DS-4 can grow rapidly in an organic carbon source heterotrophic culture medium, and after the culture is carried out for 30 hours, the OD value of a bacterial liquid reaches 0.99, which is far higher than the result of the culture in an inorganic environment. This indicates that the nutritional type of the bacterium is facultative; in engineering application, the organic carbon source heterotrophic culture medium can be used for quickly obtaining high-density thalli.
Example 3: desulfurization bottle test
200ml of sodium sulfide medium solution (600 mg/LS)2-) Sterilizing under ultraviolet for 30 min, aseptically packaging into sterilized 500ml triangular flask, adding 50ml Sphingobacterium bacteria solution for 30 hr, plugging the flask with aerobic plug, shaking in shaking table (30 deg.C, 200r/min), and measuring initial sulfide concentration to be 460mg/L S2-(ii) a The control group is added with the same amount of sterile water, each experimental group is provided with three repetitions, samples are taken every 30 minutes to determine the sulfide concentration and the elemental sulfur concentration, and the results are averaged. After 5 hours, the measurement calculation results are shown in FIG. 3. After the desulfurization bacteria are added into the sodium sulfide solution of an experimental group, the concentration of sulfide is obviously reduced compared with that of illumination, the conversion rate of elemental sulfur is obviously higher, wherein the removal rate of sulfide (a) reaches 99.13 percent, and the elemental sulfur isThe cumulative concentration (b) reached 358.49mg/L, corresponding to an elemental sulphur conversion of 77.9%. The removal of sulfide in the control group was due to shaking the flask to promote sulfide evolution, but as can be seen from elemental sulfur conversion, sulfide removal was purely physical and not biotransformation.
Example 4: devulcanizer test
5L of sodium sulfide medium solution (designed to have an initial concentration of 1200mg/L S)2-Actually measured concentration of 1098.75mg/L) is filled into a sterilized 8L reactor (the reactor is added with soft filler for biofilm culturing of microorganisms), then 500ml of sphingosine bacillus liquid for 30h of culture is added, aeration is carried out by an air pump, the aeration rate is 0.4L/min, the environmental temperature is kept at 28-30 ℃, biofilm culturing is carried out for 7 days under the condition, microorganisms can be attached to the filler, and then a verification test for removing sulfide is carried out; after the bacteria successfully form a membrane in the reactor, adding 1000ml of a sodium sulfide culture medium solution which is sterilized by ultraviolet into the reactor; the control group is added with the same amount of sterile water, each experimental group is provided with three repetitions, samples are taken every 30 minutes to determine the sulfide concentration and the elemental sulfur concentration, and the average values are taken. After 3.5h, the measurement calculation results are shown in the following table 1.
TABLE 1 Sphingobacterium pot desulfurization Effect
From the above table, after the sodium sulfide solution is added into the reactor after the film formation of the experimental group for 3.5 hours, the removal rate of sulfide is obviously higher than that of the control group, and the conversion rate of elemental sulfur is obviously higher, wherein the removal rate of sulfide can reach 98.03%, and the conversion rate of elemental sulfur can reach 66.8%. Compared with example 3, in the reactor with successful biofilm formation, the strain has higher removal rate and efficiency on sulfide, and can be applied to desulfurization application. The removal of sulfide in the control group was due to the evolution of sulfide by air stripping, but as can be seen from elemental sulfur conversion, sulfide removal was purely physical and not bioconversion.
Example 5: biological sulfur particle observation
Scanning electron microscope observation is carried out on biological sulfur particles formed by polymerization of elemental sulfur in wastewater treated by the Sphingobacterium DS-4, and the result is shown in the attached figure 4: the biological sulfur particles in the wastewater have different shapes and sizes, and the surfaces of the particles are provided with bulges.
Sequence listing
<110> institute of biological research of Chengdu of Chinese academy of sciences
<120> sphingosine desulfurization bacterium and application thereof
<160>1
<210>1
<211>878
<212>DNA
<213> Sphingobacterium sp
<400>1
gaaggatggc gtctgtctac aatgcagtcg agcgcaccct tcggggtgct cggtgtccgg 60
gtgaatagtg cgtgacggta ggccctttgg tgcgcaatct acccgtatca cggggatagc 120
cccagtatgc gcccttcggg gcaaagattt atctcccccg gcatccccgc gttggattaa 180
gttattggag gggtatgggc tcacgtgacc taagatcctt ggtggtttga gaggatgatc 240
agcgaacatg tcactgagat ctgaccagac tcctaccaca cgggtactgg gacaatctta 300
aaactcctac gcaaccctga tctaacgatg cctggtgaat gatgacagcc ctgaacctgc 360
catgctcttt gcacgatgac tgccctatgg gttgtaaact gcttttgtta ggctaactcc 420
ccgtctacat gtacgtggct gaatgtaccg ctagcgttgt gctcaattac tggccggcca 480
gcgcacgtag taataccgaa gatccgagcg ttatccggat ttattgggtt taagaggtgc 540
ctacgcggta ctttaagtcg agggtgagag actgcgtgtt attgtcgtgt gcctttgata 600
ctgaagtgct tgaatgcaat acaagacggc ggagtgagac actgactcga tacatgcata 660
gaagtgtctc agaatgccga ttgcgaaagg atctgtctac tgcgttactg acccctgatg 720
caaaaaatgc tggtcatcga acagcatgat gtacccttga cagaccctac ccctataaag 780
atgacaactc cgatgattgc accctacccg atcacatccc aaacgaaatc gataagtttc 840
tccatcttgg cgagggcgct tcgtgcgttt tgtattta 878
Claims (3)
1. A sphingosine desulfurization (Sphingobacterium sp) DS-4 strain, which is characterized by the following preservation number: CGMCC No.12632, which is short rod-shaped, 1.2-2 μm long, 0.8-0.1.2 μm wide, gram-negative and has no flagella; the growth pH is 5.0-9.0; the growth temperature is 20-35 ℃; the bacterial colony is oval, the surface is concave, and the color is milky white; the nutrition type of the strain is facultative, and the strain can utilize organic carbon sources such as beef extract, peptone and yeast powder to perform heterotrophic rapid growth and can also utilize carbon dioxide as a unique carbon source to perform autotrophic growth; when the self-curing process is carried out by taking carbon dioxide as the only carbon source, the bacteria can utilize oxygen to oxidize sulfides into elemental sulfur or sulfates.
2. The desulfospironobacter of claim 1, wherein the 16S rDNA sequence of the sphingosine bacterium is set forth in SEQ ID No. 1.
3. Use of the bacterium sphingosine desulfurated according to claim 1 for sulfur compound desulfurization.
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| CN104357367A (en) * | 2014-11-24 | 2015-02-18 | 中国烟草总公司郑州烟草研究院 | Strain capable of degrading di(2-ethylhexyl) phthalate and application of strain to tobacco planting |
| CN104593347A (en) * | 2015-03-05 | 2015-05-06 | 深圳市海普瑞药业股份有限公司 | Heparinases obtained from Sphingobacterium daejeonense as well as preparation and application thereof |
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