Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a micro-oxysulfur micro-spirillum and application thereof in biological desulfurization, in particular to halophilic basophilic micro-oxysulfur micro-spirillum with high elemental sulfur yield and application thereof in biological desulfurization.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides a halophilic basophilic micro oxysulfuricus bacterium for high elemental sulfur yield, which is named as micro oxysulfuricus bacterium (Thiomicrospira microaerophila) BDL05 strain, the preservation unit is China general microbiological culture Collection center, the preservation time is 2020, 6, month and 10 days, the preservation number is CGMCC No.20060, and the address is as follows: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North.
The high-yield elemental sulfur refers to that when the sulfur-containing gas or wastewater is treated by the sulfur oxysulfide micro-spirochete, the generation rate of elemental sulfur is more than 95%.
The microaerophilic sulfur-micro spirillum BDL05 strain related by the invention is separated from the sediment at the bank of the desert saline-alkali lake of inner Mongolia badam and has the characteristic of oxidizing sulfide to generate elemental sulfur under the conditions of high salt and high alkali. The strain is chemoautotrophic microorganism, the cell energy is derived from chemical energy generated by oxidation of reduced sulfur-containing compounds such as hydrosulfide, elemental sulfur, sodium thiosulfate, etc., and the carbon source is derived from carbon dioxide in atmosphere, and can be pH8.0-12.0, 0.5-4.0M Na+And the growth is carried out under the conditions of high salt and high alkali.
The invention relates to a method for separating and culturing a Microspirillum oxysulfide BDL05 strain, which comprises the following steps:
(1) collecting sediments beside a saline-alkali lake of a desert of inner Mongolia badan Jilin, putting 1g of the sediments in a 100mL liquid culture medium of sodium thiosulfate, and carrying out shaking culture on a shaking table at 30 ℃ and 150rpm for 60 hours;
(2) inoculating 1mL of the suspension into 100mL of sodium thiosulfate liquid culture medium, and continuing shaking culture at 30 ℃ and 150rpm for 60 hours by a shaking table;
(3) coating 100 mu L of suspension on the surface of a sodium thiosulfate solid culture medium, standing and culturing at 30 ℃ for 60h, and selecting white colonies for re-screening and identification;
wherein the sodium thiosulfate liquid medium includes, for exampleThe following components: 30g/L Na2S2O3、30g/L NaHCO3、5g/L NaOH、10g/L NaCl、0.5g/L NH4Cl、0.5g/L KNO3、2.0g/L K2HPO4、0.1g/L MgCl2。
Wherein, the other components in the sodium thiosulfate solid culture medium are as follows: 30g/L Na2S2O3、30g/L NaHCO3、5g/L NaOH、10g/L NaCl、0.5g/L NH4Cl、0.5g/L KNO3、2.0g/L K2HPO4、0.1g/L MgCl21.5% agar powder;
after multiple purifications, a micro-aerobic sulfur micro-spirillum BDL05 capable of producing elemental sulfur is obtained.
In a second aspect, the invention provides a method for culturing high-yield elemental sulfur-producing halophilic microspiromyces oxysporum, which comprises culturing the high-yield elemental sulfur-producing halophilic microspiromyces oxysporum in a medium containing sodium thiosulfate.
Preferably, the sodium thiosulfate-containing medium comprises Na2S2O3、NaHCO3、NH4Cl、KNO3、K2HPO4And MgCl2。
Preferably, the medium containing sodium thiosulfate also comprises NaOH and/or NaCl.
Preferably, the culture medium containing sodium thiosulfate comprises 10-30g/L of Na in mass concentration2S2O3、30-60g/L NaHCO3、0-30g/L NaOH、0-100g/L NaCl、0.1-1.0g/L NH4Cl、0.1-1.0g/L KNO3、0.5-5.0g/L K2HPO4And 0.05-0.3g/L MgCl2。
The Na is2S2O3The mass concentration of the organic acid can be 10g/L, 15g/L, 20g/L, 25g/L or 30g/L, and the like, and specific point values in the numerical value range can be selected, so that the description is omitted.
Said NaHCO3The mass concentration of (b) can be 30g/L, 35g/L, 40g/L, 45g/L, 50g/L, 55g/L or 60g/L, etcThe specific point values in the range can be selected, and are not described in detail herein.
The mass concentration of the NaOH can be 5g/L, 10g/L, 15g/L, 20g/L, 25g/L or 30g/L, and the like, specific point values in the numerical value range can be selected, and are not repeated here, and when the mass concentration is 0, the NaOH is not added.
The mass concentration of NaCl can be 5g/L, 10g/L, 20g/L, 30g/L, 40g/L, 50g/L, 60g/L, 80g/L or 100g/L, and the like, specific point values in the numerical value range can be selected, and are not repeated one by one, and when the mass concentration is 0, NaCl is not added.
The NH4The mass concentration of Cl can be 0.1g/L, 0.2g/L, 0.3g/L, 0.4g/L, 0.5g/L, 0.6g/L, 0.7g/L, 0.9g/L or 1.0g/L, and the like, and specific point values in the numerical value range can be selected, so that the details are not repeated.
The KNO3The mass concentration of the organic acid can be 0.1g/L, 0.2g/L, 0.3g/L, 0.4g/L, 0.5g/L, 0.6g/L, 0.7g/L, 0.9g/L or 1.0g/L, and the like, and specific point values in the numerical value range can be selected, so that the description is omitted.
Said K2HPO4The mass concentration of the organic acid can be 0.5g/L, 0.6g/L, 0.8g/L, 1.0g/L, 1.5g/L, 2.0g/L, 3.0g/L, 4.0g/L or 5.0g/L, and the like, and specific point values in the numerical value range can be selected, so that the description is omitted.
Said MgCl2The mass concentration of the organic acid can be 0.05g/L, 0.1g/L, 0.15g/L, 0.2g/L, 0.25g/L or 0.3g/L, and the like, and specific point values in the numerical value range can be selected, so that the repeated description is omitted.
Preferably, the pH of the medium containing sodium thiosulfate is 8.0 to 12.0, for example, 8.0, 9.0, 10.0, 11.0 or 12.0, and the sodium ion concentration is 0.5 to 4.0M, for example, 0.5M, 1.0M, 1.5M, 2.0M, 2.5M, 3.0M or 4.0M, and the specific values in the above numerical range can be selected, which is not repeated herein.
In a third aspect, the invention provides an application of halophilic basophilic micro sulfur-oxygen micro spirillum with high elemental sulfur yield in biological desulfurization of hydrogen sulfide-containing gas.
In the present invention, the method of application comprises the steps of:
(1) absorbing hydrogen sulfide in the gas by using an alkaline washing solution to form a high-sulfur solution;
(2) and (2) inoculating the halophilic basophilic micro oxysulfur micro spirillum with high elemental sulfur yield into the high sulfur-containing solution obtained in the step (1) for desulfurization treatment.
In the desulfurization treatment process, sodium hydrosulfide is oxidized into elemental sulfur by microaspirillum oxysulfide to generate sodium hydroxide; the elemental sulfur is agglomerated to form micron-sized sulfur particles, naturally settled for 0.1min-2h and then discharged from the bioreactor, so as to achieve the aim of desulfurization; the lye discharged from the sulfur particles will continue to be recycled for absorption of hydrogen sulfide in the gas. The generation rate of elemental sulfur is more than 95 percent, and the removal rate of hydrogen sulfide reaches more than 99 percent.
Preferably, the gas comprises natural gas, biogas, petroleum refinery gas, coal refinery gas, landfill gas or viscose production waste gas.
Preferably, the alkaline washing solution contains any one or a combination of at least two of sodium carbonate, sodium bicarbonate or sodium hydroxide; the combination of at least two of the above-mentioned compounds, such as the combination of sodium carbonate and sodium bicarbonate, the combination of sodium bicarbonate and sodium hydroxide, etc., can be selected in any other combination manner, and thus, the details are not repeated herein.
Preferably, the pH of the alkaline washing solution is 8.0-12.0, for example, 8.0, 9.0, 10.0, 11.0 or 12.0, and the sodium ion concentration is 0.5-4.0M, for example, 0.5M, 1.0M, 1.5M, 2.0M, 2.5M, 3.0M or 4.0M, and the specific values in the above-mentioned value range can be selected, which is not repeated herein.
Preferably, the concentration of sodium hydrosulfide in the high sulfur-containing solution is 1.0-3.0g/L, such as 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L or 3.0g/L, and the like, and specific values within the above numerical value range can be selected, and are not repeated herein.
Preferably, the inoculation amount of the halophilic basophilic microspiromyces for high yield of elemental sulfur is 1-5%, for example, 1%, 2%, 3%, 4%, or 5%, and the specific values in the above numerical range can be selected, and are not described in detail herein.
In a fourth aspect, the invention provides an application of the halophilic basophilic micro sulfur-oxygen micro spirillum with high elemental sulfur yield in biological desulfurization of alkaline wastewater containing sulfide.
In the present invention, the method of application comprises the steps of:
the halophilic alkalophilic micro-oxysulfurin microspira bacteria with high elemental sulfur yield are inoculated into alkaline wastewater containing sulfide for desulfurization treatment.
In the desulfurization treatment process, the sulfur compound is oxidized into elemental sulfur by micro sulfur-oxidizing micro spirillum, and meanwhile, alkali is regenerated; the elemental sulfur is agglomerated to form micron-sized sulfur particles, naturally settled for 0.1min-2h and then discharged from the bioreactor, so that the aim of desulfurization is fulfilled, and the generation rate of the elemental sulfur is more than 95%.
Preferably, the sulfide-containing alkaline wastewater comprises leather wastewater, petroleum refining wastewater, pesticide production wastewater or synthetic ammonia wastewater.
Preferably, the pH of the sulfide-containing alkaline wastewater is 8.0 to 12.0, for example, 8.0, 9.0, 10.0, 11.0 or 12.0, and the concentration of sodium ions is 0.5 to 4.0M, for example, 0.5M, 1.0M, 1.5M, 2.0M, 2.5M, 3.0M or 4.0M, and the like, and specific values within the above-mentioned value range can be selected, which is not described in detail herein.
Preferably, the concentration of sulfide in the sulfide-containing alkaline wastewater is 0.1-3.0g/L, such as 0.1g/L, 0.5g/L, 1.0g/L, 2.0g/L, 2.5g/L or 3.0g/L, and the like, and specific values in the above numerical value range can be selected, and are not repeated herein.
Preferably, the inoculation amount of the halophilic basophilic microspiromyces for high yield of elemental sulfur is 1-5%, for example, 1%, 2%, 3%, 4%, or 5%, and the specific values in the above numerical range can be selected, and are not described in detail herein.
Preferably, the oxidation-reduction potential of the sulfide-containing alkaline wastewater is-150 mV to-400 mV, such as-150 mV, -200mV, -250mV, -300mV, -350mV or-400 mV, and the like, and specific values in the above numerical value ranges can be selected, and are not repeated herein.
Preferably, the adjustment of the oxidation-reduction potential of the sulfide-containing alkaline wastewater comprises adjusting the supply of oxygen.
Compared with the prior art, the invention has the following beneficial effects:
the halophilic basophilic micro oxysulfuricus (Thiomicrospira microaerophila BDL05) capable of producing the elemental sulfur at high yield can oxidize hydrosulfide to generate the elemental sulfur under the condition of high salt and high alkali, the yield of the elemental sulfur reaches more than 95%, the stability of a biological desulfurization system is remarkably improved, and the application range of the biological desulfurization technology is expanded.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
Example 1
The present example provides a method for screening and separating halophilic basophilic microspira BDL05 with high elemental sulfur yield, which is performed as follows:
(1) collecting sediments beside a saline-alkali lake of a desert of inner Mongolia badan Jilin, putting 1g of the sediments in a 100mL liquid culture medium of sodium thiosulfate, and carrying out shaking culture on a shaking table at 30 ℃ and 150rpm for 60 hours;
(2) inoculating 1mL of the suspension into 100mL of sodium thiosulfate liquid culture medium, and continuing shaking culture at 30 ℃ and 150rpm for 60 hours by a shaking table;
(3) coating 100 mu L of suspension on the surface of a sodium thiosulfate solid culture medium, standing and culturing at 30 ℃ for 60h, and selecting white colonies for re-screening and identification;
wherein, the sodium thiosulfate liquid culture medium comprises the following components: 30g/L Na2S2O3、30g/L NaHCO3、5g/L NaOH、10g/L NaCl、0.5g/L NH4Cl、0.5g/L KNO3、2.0g/L K2HPO4、0.1g/L MgCl2。
Wherein, the other components in the sodium thiosulfate solid culture medium are as follows: 30g/L Na2S2O3、30g/L NaHCO3、5g/L NaOH、10g/L NaCl、0.5g/L NH4Cl、0.5g/L KNO3、2.0g/L K2HPO4、0.1g/L MgCl21.5% agar powder;
after multiple purifications, a micro-aerobic sulfur micro-spirillum BDL05 capable of producing elemental sulfur is obtained.
The colony morphology of the obtained microaspirillum BDL05 was observed, as shown in fig. 1: the colony is white, round-like and irregular in edge, and the white colony gradually becomes transparent in the later culture period.
The obtained Microspirillum oxysporum BDL05 was gram-stained and negative, as shown in FIG. 2. The evolutionary position of Microspirillum oxysulphatum BDL05 is shown in FIG. 3.
The obtained Microspirillum microaerophilum BDL05 was subjected to 16S rDNA sequence determination, and the sequence was as follows:
GAAGTGCGCAGCTACCATGCAGTCGAACGGTAACAGAAGAGCTTGCTCTTGCTGACGAGTGGCGGACGGGTGAGTAATGCATAGGAATCTGCCCTCTAGTTGGGGATACCGTAGGGAAACTTACGTTAATACCGAATAGTCTCTAAGGAGTAAAGGTGGCCTCTACTTGTAAGCTATCGCTAGAGGATGAGCCTATGTTAGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCTATAGCTGGTTTGAGAGGATGATCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGCCCGAGGGTTGTAAAGCACTTTTAGCGAGGAGGAAAGGATGTAGATTAATACCCTGCATCTGTGACGTTACTCGCAGAAAAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGTAATACAGAGGGTGCAAGCGTTATTCGGAATTACTGGGCGTAAAGCGCGCGTAGGCGGATTGTTAAGTCAGTTGTGAAAGCCCTGGGCTCAACCTAGGAACGGCGATTGAAACTGGCAATCTAGAGTTTAGTAGAGGGAAGGGGAATTTCTGGAGTAGCAGTGAAATGCGTAGATATCAGAAGGAACATCAGTGGCGAAGGCGCCTTCCTGGACTAAAACTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAACGGGATTAGATACCCCGGTAGTCCACGCCCTAAACGATGTCAACTAGCTGTTGGTCTTATTAAAAAGATTAGTAGCGCAGCTAACGCGATAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCATCCCTTGACATCCACAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAGCTGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATCATTAG。
the above analysis results indicate that it is a Thiomicrospira microaerophila (Thiomicrospira microaerophila).
Example 2
This example explores the characteristics of the growth of Microspirillum oxysporum BDL05 using sodium thiosulfate and operates as follows:
after streak activation culture of the strain of the sulfur micro spirillum BDL05 preserved at 4 ℃, inoculating the strain into 100mL of sodium thiosulfate liquid culture medium, placing the strain in a shaking table at 30 ℃ and 160r/min for 30h, and entering a logarithmic phase to be used as seed liquid for later use. Inoculating the strain in 100mL of sodium thiosulfate liquid medium at an inoculation amount of 2%, sampling at regular intervals, and determining S2O3 2-Concentration, SO4 2-Concentration and OD values were varied with time.
Wherein, the sodium thiosulfate liquid culture medium comprises the following components: 30g/L Na2S2O3、30g/L NaHCO3、5g/L NaOH、10g/L NaCl、0.5g/L NH4Cl、0.5g/L KNO3、2.0g/L K2HPO4、0.1g/L MgCl2。
Growth curves and S of the culture of Strain BDL052O3 2-And SO4 2-The change in concentration is shown in FIG. 4, and it can be seen from FIG. 4 that: the growth of the strain is slower in the first 7h under the conditions of 30 ℃ and 160 r/min; after the adaptation adjustment period, the strain BDL05 rapidly grows, the OD of the thallus of the culture solution reaches 0.6 at 32h, and then the OD value begins to decrease; s2O3 2-The concentration was initially 44mmol/L, S as the incubation time was extended2O3 2-The concentration is continuously reduced and is reduced to 0.85mmol/L in 32 h; SO (SO)4 2-The concentration gradually increases with the culture time, and SO is added until the concentration reaches 40h4 2-The concentration reaches 70.3 mmol/L. The sulfur utilization rate was 1.34 mmol/L/h.
It was described that BDL05 can be grown using sodium thiosulfate, and the above sodium thiosulfate liquid medium can be used as a culture solution for culturing BDL 05.
Example 3
This example uses Microspirillum oxysulphide BDL05 for the treatment of a hydrogen sulphide-containing gas, operating as follows:
(1) a test hydrogen sulfide-containing gas containing 6000ppm hydrogen sulfide and the balance nitrogen was prepared.
(2) The gas containing hydrogen sulfide flows through an absorption tower in a countercurrent way, during the process, the gas is contacted with alkali liquor, wherein the hydrogen sulfide is absorbed by the alkali liquor to form high-sulfur solution containing high sodium hydrosulfide; the high-sulfur solution enters a bioreactor; in a bioreactor, oxidizing sulfur hydride into elemental sulfur by microaspirillum BDL05 by controlling oxidation-reduction potential, and simultaneously regenerating alkali liquor for recycling;
wherein the alkali solution is an aqueous solution containing sodium carbonate, sodium bicarbonate or sodium hydroxide, the pH value is 9.7, and the sodium ion concentration is 3.0M; the concentration of sodium hydrosulfide in the high-sulfur solution is 2 g/L. In the biological sulfur oxidation process, the oxidation-reduction potential is controlled to be about-380 mv by controlling the sulfur oxidation products through the aeration quantity.
Through calculation, the generation rate of the elemental sulfur reaches more than 95 percent; the content of hydrogen sulfide in the purified gas is lower than 10ppm, and the removal rate of the hydrogen sulfide reaches more than 99%. The micro sulfur oxysulfide micro spirillum BDL05 has high-efficiency sulfur oxidizing capability, and the indexes of elemental sulfur generation rate, hydrogen sulfide removal rate and the like reach or exceed the existing strains, and can be applied to the treatment of the gas containing hydrogen sulfide.
Example 4
In this example, the operation of using the Microspirillum oxysulfide BDL05 for the treatment of alkaline sulfide wastewater is as follows:
(1) the wastewater for the preparation test is wastewater of a depilation section of a tanning enterprise, wherein the sodium sulfide reaches 2500mg/L, the pH value is 11.0, and the sodium ion concentration reaches 2.5M.
(2) Filtering the wastewater to remove substances such as large fat and the like; then adding the microspiromyces oxysulfuricus BDL05 seed liquid into sulfur-containing wastewater in an inoculation amount of 5%; by regulating and controlling aeration quantity and controlling the oxidation-reduction potential to be about-350 mv, the sulfur hydride is oxidized into elemental sulfur by the microaspirillum BDL05, and the elemental sulfur is separated from the wastewater through the sedimentation effect. The experiment is continued for 15 days, and the sulfide concentration and the elemental sulfur yield in the wastewater are uninterruptedly determined.
As shown in FIG. 5, the elemental sulfur yield was 95% or more; the content of sodium sulfide in the purified wastewater is lower than 25mg/L, and the removal rate of the sodium sulfide reaches more than 99%. The micro sulfur oxysulfide micro spirillum BDL05 has high-efficiency sulfur oxidizing capability, and the indexes of elemental sulfur generation rate, sulfide removal rate and the like reach or exceed the prior strains, and can be applied to the treatment of the wastewater containing the sulfide.
The applicant states that the present invention is illustrated by the above examples, but the present invention is not limited to the above examples, i.e. it does not mean that the present invention must rely on the above examples to be implemented. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
SEQUENCE LISTING
<110> institute of Process engineering of Chinese academy of sciences
<120> halophilic basophilic micro oxysulfuricus bacterium for high yield of elemental sulfur and application thereof in biological desulfurization
<130> 2020
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1084
<212> DNA
<213> Microspira microaerophila (Thiomicrospira microaerophila) BDL05
<400> 1
gaagtgcgca gctaccatgc agtcgaacgg taacagaaga gcttgctctt gctgacgagt 60
ggcggacggg tgagtaatgc ataggaatct gccctctagt tggggatacc gtagggaaac 120
ttacgttaat accgaatagt ctctaaggag taaaggtggc ctctacttgt aagctatcgc 180
tagaggatga gcctatgtta gattagctag ttggtgaggt aatggctcac caaggcgacg 240
atctatagct ggtttgagag gatgatcagc cacactggga ctgagacacg gcccagactc 300
ctacgggagg cagcagtggg gaatattgga caatgggcgc aagcctgatc cagccatgcc 360
gcgtgtgtga agaaggcccg agggttgtaa agcactttta gcgaggagga aaggatgtag 420
attaataccc tgcatctgtg acgttactcg cagaaaaagc accggctaac tctgtgccag 480
cagccgcggt aatacagagg gtgcaagcgt tattcggaat tactgggcgt aaagcgcgcg 540
taggcggatt gttaagtcag ttgtgaaagc cctgggctca acctaggaac ggcgattgaa 600
actggcaatc tagagtttag tagagggaag gggaatttct ggagtagcag tgaaatgcgt 660
agatatcaga aggaacatca gtggcgaagg cgccttcctg gactaaaact gacgctgagg 720
tgcgaaagcg tggggagcaa acgggattag ataccccggt agtccacgcc ctaaacgatg 780
tcaactagct gttggtctta ttaaaaagat tagtagcgca gctaacgcga taagttgacc 840
gcctggggag tacggtcgca agattaaaac tcaaaggaat tgacgggggc ccgcacaagc 900
ggtggagcat gtggtttaat tcgatgcaac gcgaagaacc ttaccatccc ttgacatcca 960
cagaactttc cagagatgga ttggtgcctt cgggagctgt gagacaggtg ctgcatggct 1020
gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccctatca 1080
ttag 1084