A chemoautotrophic Annwoodia sulfur-oxidizing bacteria strain and its application

Abstract

The invention discloses a chemoautotrophic plant Annwoodia The invention relates to a sulfur oxidizing bacteria strain and its application, belonging to the technical field of microorganisms and environmental protection. The strain is named Annwoodia sp. C03, with a deposit number of GDMCC No: 65042. Thiobacillaceae Corey Annwoodia A new species of the genus was added Annwoodia The strain C03 can bio-oxidize sulfide, elemental sulfur, thiosulfate, thiocyanate, sulfite, tetrathionate and other reduced sulfides under aerobic conditions. In addition, the strain C03 can also simultaneously carry out sulfur oxidation and denitrification under hypoxic conditions, that is, the strain has good application value in the treatment of wastewater containing reduced sulfides.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and environmental protection, and particularly relates to a chemoautotrophic Annwoodia The invention relates to a sulfur oxidizing bacteria strain and application thereof. Background Art

[0002] Sulfur is an important non-metallic element, widely present in natural environments such as ore bodies, rocks, water bodies, and the atmosphere. It is also one of the important elements that constitute organic life. It has rich chemical properties and a wide range of applications. It plays an important role in nature, organisms, and industrial production. The outermost electron number of sulfur atoms is 6. In chemical reactions, it can form -2 to +6 ion forms by gaining and losing electrons. Among them, low-valence reduced sulfides are a type of environmental pollutants that cannot be ignored. Most of them are toxic and highly corrosive, and have great environmental damage. Among the environmental damage caused by reduced sulfides, acid rain is undoubtedly one of the most well-known forms of harm. With the increasing scale of global industrialization and the increase in population, the total amount of reduced sulfides emitted by humans in the process of production and life has increased rapidly. In addition, the concept of green development has been increasingly valued, which has put forward new requirements for development. Traditional treatment methods such as physical methods and chemical methods can no longer meet the growing demand for reduced sulfide treatment. Compared with traditional physical and chemical methods, biological method is a green and environmentally friendly treatment method. It uses the metabolic ability of microorganisms on reduced sulfides to convert harmful reduced sulfides into non-toxic sulfates. It has the advantages of environmental friendliness, mild treatment conditions, simple operation, safety, efficiency, sustainability, etc. in treating pollutants such as reduced sulfides, showing broad application prospects and great application value.

[0003] Sulfur oxidizing bacteria (SOB) play a core role in the biological treatment of reduced sulfide, and chemoautotrophic sulfur oxidizing bacteria are a type of sulfur oxidizing bacteria that can oxidize reduced sulfide into high-valent sulfur-containing compounds, and produce enough energy to fix carbon dioxide during the sulfur oxidation process, synthesize organic matter to meet their own growth metabolism, and do not rely on exogenous organic matter. Chemoautotrophic sulfur oxidizing bacteria are widely present in nature and play an important role in biogeochemical cycles, such as sulfur cycle, carbon cycle, and nitrogen cycle. They have unique ecological functions and application values ​​in ecosystems.

[0004] Therefore, isolating and identifying new chemoautotrophic sulfur-oxidizing strains from the environment and studying the sulfur-oxidizing characteristics of the strains and their applications can deepen our understanding of chemoautotrophic sulfur-oxidizing bacteria, explore the application value of chemoautotrophic sulfur-oxidizing bacteria, promote the application of chemoautotrophic sulfur-oxidizing bacteria, and provide new microbial resources with application value for environmental governance and ecological protection. Summary of the invention

[0005] In order to overcome the shortcomings and deficiencies of the prior art, the present invention aims to provide a chemoautotrophic Annwoodia Belongs to the sulfur oxidizing bacteria strain.

[0006] Another object of the present invention is to provide the above-mentioned chemoautotrophic Annwoodia Application of sulfur oxidizing bacteria strains.

[0007] The purpose of the present invention is achieved through the following technical solutions:

[0008] A chemoautotrophic plant Annwoodia It is a sulfur oxidizing bacteria strain named Annwoodia sp. C03, with the accession number GDMCC No: 65042, was deposited on August 28, 2024 at the Guangdong Microbial Culture Collection Center, Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou, Guangdong, China.

[0009] Chemoautotrophic Annwoodia Application of sulfur-oxidizing bacterial strains in the redox sulfide process.

[0010] Furthermore, the application comprises the following steps:

[0011] The above chemoautotrophic Annwoodia The sulfur oxidizing bacteria strain is placed in an environment containing reduced sulfide for growth. Furthermore, the environment includes at least one of water and soil.

[0012] Furthermore, the redox sulfide is obtained by oxidizing the reducing sulfide into sulfate.

[0013] Further, the reducing sulfide includes at least one of elemental sulfur, thiosulfate, alkali metal sulfide, tetrathionate, thiocyanate and sulfite; further, the thiosulfate includes sodium thiosulfate, the alkali metal sulfide includes sodium sulfide, the tetrathionate includes sodium tetrathionate, the thiocyanate includes sodium thiocyanate, and the sulfite includes sodium sulfite.

[0014] Furthermore, the growth is performed at an oxygen concentration of 0.1% to 21%; further, the growth is performed at an oxygen concentration of 1% to 21%; further, the growth is performed at an oxygen concentration of 3% to 21%.

[0015] Furthermore, the growth temperature is 25-35°C; furthermore, 30-35°C; furthermore, 30°C.

[0016] Chemoautotrophic AnnwoodiaThe invention relates to the application of sulfur oxidizing bacteria strains in oxidizing reduced sulfide and achieving simultaneous denitrification under hypoxic conditions.

[0017] Furthermore, the application comprises the following steps:

[0018] The above chemoautotrophic Annwoodia The sulfur oxidizing bacteria strain is placed in a low oxygen environment containing reducing sulfide and inorganic nitrogen source for growth. Furthermore, the environment includes at least one of water and soil.

[0019] Furthermore, the redox sulfide is obtained by oxidizing the reducing sulfide into sulfate.

[0020] Further, the reducing sulfide includes at least one of elemental sulfur, thiosulfate, alkali metal sulfide, tetrathionate, thiocyanate and sulfite; further, the thiosulfate includes sodium thiosulfate, the alkali metal sulfide includes sodium sulfide, the tetrathionate includes sodium tetrathionate, the thiocyanate includes sodium thiocyanate, and the sulfite includes sodium sulfite.

[0021] Furthermore, the denitrification is to reduce nitrogen in the inorganic nitrogen source to gaseous nitrogen; further, the gaseous nitrogen is N2O.

[0022] Furthermore, the denitrification is to reduce nitrate ions or nitrite ions to gaseous nitrogen.

[0023] Further, the inorganic nitrogen source includes at least one of nitrate and nitrite;

[0024] Furthermore, the nitrate includes sodium nitrate, and the nitrite includes sodium nitrite.

[0025] Furthermore, the hypoxia is a condition where the oxygen concentration is 0.1% to 2%, preferably a condition where the oxygen concentration is 0.1% to 1%.

[0026] Furthermore, the growth temperature is 25-35°C; furthermore, 30-35°C; furthermore, 30°C.

[0027] Compared with the prior art, the present invention has the following advantages and effects:

[0028] The present invention provides Annwoodia sp. C03 Thiobacillaceae Corey Annwoodia A new species of the genus was added AnnwoodiaThe strain C03 can bio-oxidize sulfide, elemental sulfur, thiosulfate, thiocyanate, sulfite, tetrathionate and other reduced sulfides under aerobic conditions. In addition, the strain C03 can also simultaneously carry out sulfur oxidation and denitrification under hypoxic conditions, that is, the strain has good application value in the treatment of wastewater containing reduced sulfides. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Figure 2 is the colony and cell morphology of strain C03; A is the colony morphology of strain C03 on the enrichment screening solid inorganic culture medium plate, and the culture conditions are microaerobic conditions (3% to 5% oxygen concentration), B is a photo of bacterial Gram staining observed under an optical microscope (100×), and C is a scanning electron microscope photo (25.00 k×).

[0030] Figure 2 This is a phylogenetic tree constructed based on the 16S rDNA of strain C03.

[0031] Figure 3 For different oxygen concentrations Annwoodia sp. C03 sulfur oxidation metabolic characteristics; wherein, A is the consumption result of thiosulfate in the sulfur oxidation metabolism process under different oxygen concentrations, B is the generation result of sulfate in the sulfur oxidation metabolism process under different oxygen concentrations, C is the effect of pH in the sulfur oxidation metabolism process under different oxygen concentrations, and D is the cell number result at 0 hour and 60 hours in the sulfur oxidation metabolism process under different oxygen concentrations.

[0032] Figure 4 For different oxygen concentrations Annwoodia sp. C03 on the formation of colonies on the enrichment screening solid inorganic culture medium plate; A is the effect diagram of colony formation under normal oxygen conditions, and B is the effect diagram of colony formation under microaerobic conditions.

[0033] Figure 5 for Annwoodia sp. C03 sulfur oxidation growth curve results under normal oxygen conditions; wherein, A is the consumption result of thiosulfate ion in the process of sulfur oxidation metabolism under normal oxygen conditions, B is the generation result of sulfate ion in the process of sulfur oxidation metabolism under normal oxygen conditions, C is the effect of pH in the process of sulfur oxidation metabolism under normal oxygen conditions, and D is the change result of cell number in the process of sulfur oxidation metabolism under normal oxygen conditions.

[0034] Figure 6 for Annwoodiasp. C03 sulfur oxidation and denitrification metabolic characteristics under hypoxic conditions; wherein, A is a graph showing the consumption of thiosulfate ions during sulfur oxidation and denitrification metabolism under hypoxic conditions, B is a graph showing the generation of sulfate ions during sulfur oxidation and denitrification metabolism under hypoxic conditions, C is a graph showing the consumption of nitrate nitrogen during sulfur oxidation and denitrification metabolism under hypoxic conditions, and D is a graph showing the accumulation of nitrite nitrogen during sulfur oxidation and denitrification metabolism under hypoxic conditions.

[0035] Figure 7 for Annwoodia Graph showing the results of sp. C03's degradation of thiocyanate under normal oxygen conditions. DETAILED DESCRIPTION

[0036] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0037] The experimental methods in the following examples without specific experimental conditions are usually carried out under conventional experimental conditions or the experimental conditions recommended by the manufacturer. The materials and reagents used are commercially available unless otherwise specified.

[0038] Example 1

[0039] Chemoautotrophic Annwoodia Enrichment, screening, separation and identification of sp. C03

[0040] (1) Enrichment of sulfur-oxidizing bacteria: Water samples (0-30 cm) were collected from the Daxintian Village Creek site in Guangzhou City, Guangdong Province (113°35'27.028" east longitude, 23°14'44.603" north latitude) using a square 5-point sampling method. Environmental samples were inoculated in serum bottles containing enrichment screening inorganic culture medium with a headspace oxygen concentration of 1%. The sulfate content was determined after culturing in a light-proof shaker at 30°C and 150 rpm for 120 hours. Under this condition, the transfer was repeated for 5-8 generations to obtain a stable sulfur-oxidizing bacterial community. Among them, the components of the enrichment and screening inorganic culture medium are: KH2PO4 0.4g / L, K2HPO4 0.11g / L, MgCl2 0.15g / L, CaCl2·2H2O 0.15g / L, NaCl 0.75g / L, Na2S2O3·5H2O 0.75g / L, NaHCO3 0.5g / L, NH4Cl 0.11g / L, FeCl2·4H2O 0.008g / L, and trace element solution 0.5mL / L. The composition of the trace element solution is as follows: MnCl2·4H2O 0.5 g / L, CoCl2·6H2O 0.1 g / L, ZnCl2 0.1 g / L, CuCl2·2H2O 0.02 g / L, AlK(SO4)2 0.01 g / L, H3BO3 0.01 g / L, Na2MoO4·2H2O 0.1 g / L, NiCl2·6H2O 0.03 g / L, Na2WO4·2H2O 0.03 g / L, and the solvent is deionized water.

[0041] (2) Isolation of single colonies: The sulfur-oxidizing bacterial flora that has undergone multiple rounds of passages is diluted with sterile water and inoculated into an enrichment screening solid inorganic medium (15-20 g / L of agar is added to the enrichment screening inorganic medium). After culturing at 30°C under microaerobic conditions (3%-5% oxygen concentration) for 5-6 days, the growth of colonies is observed, and colonies with different colors, edges, transparency and other appearances are selected for further separation and purification. Screening by aerobic sulfur oxidation, the specific steps are as follows:

[0042] Aerobic sulfur oxidation experiment: Inoculate a single colony into a 100mL conical flask containing 50mL of enriched screening inorganic culture medium, with an oxygen concentration of 3%, and culture in a shaker at 30°C and 150rpm. Take samples every 24 hours to determine the thiosulfate content, sulfate content, pH and cell number. The aerobic sulfur oxidation experiment is judged based on the decrease in thiosulfate content, increase in sulfate content, decrease in pH and increase in cell number.

[0043] A strain was obtained through screening and named C03. In the aerobic sulfur oxidation experiment, as the culture time increased, the thiosulfate content continued to decrease, the sulfate content continued to increase, the pH continued to decrease, and the cells also proliferated rapidly. The colony morphology and cell morphology of the C03 strain are shown in Figure 2. Figure 1 The specific morphological and physiological and biochemical characteristics are described in Table 1.

[0044] Table 1 Colony morphology and physiological and biochemical characteristics of strain C03

[0045]

[0046] Note: "+": positive; "-": negative. When detecting available metabolic substrates and carbon sources, the culture medium used is basically the same as the enrichment screening inorganic culture medium, the difference is that the substances in the table are used to replace the NaHCO3 or Na2S2O3·5H2O in the enrichment screening inorganic culture medium, and the liquid culture is carried out under normal oxygen (21%) conditions; the composition of LB (Luria-Bertani) medium is: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, pH adjusted to 6.5-7.0, and the solvent is deionized water; the composition of MH (Müller-Hinton) medium is: beef extract powder 300g / L, soluble starch 1.5g / L, casein acid hydrolyzate 17.5g / L, pH adjusted to 6.5-7.0, and the solvent is deionized water; MRS (De Man, Rogosa The composition of the medium is as follows: casein digest 10.0 g / L, beef extract powder 10 g / L, yeast extract powder 4 g / L, glucose 20 g / L, Tween-80 1.08 g / L, K2HPO4 2 g / L, sodium acetate 5 g / L, ammonium citrate tribasic 2 g / L, MgSO4·7H2O 0.2 g / L, MnSO4·4H2O 0.05 g / L, L-cysteine ​​0.5 g / L, pH was adjusted to 6.5-7.0, and the solvent was deionized water.

[0047] (3) Cloning and identification of 16S rDNA of strain C03: The 16S rDNA fragment of the isolated strain C03 was amplified by colony PCR. The isolated strain C03 was inoculated into an enrichment and screening inorganic culture medium and then centrifuged to harvest the bacteria. The genomic DNA of the bacteria was extracted using the TIANamp Bacteria DNA Kit bacterial genomic DNA extraction kit. The extracted DNA was then used as a template for PCR amplification using 16S rDNA universal primers 27F and 1492R. The PCR reaction system was: 1 μL genomic DNA, 1 μL upstream primer 27F (10 μmol / L), 1 μL downstream primer 1492R (10 μmol / L), 12.5 μL DreamTaq Green PCR Master Mix (2×), and ddH2O was added to 25 μL. The PCR reaction conditions were as follows: 94°C pre-denaturation for 5 min; 94°C denaturation for 30 s, 54°C annealing for 30 s, 72°C extension for 90 s, 30 cycles; 72°C extension for 10 min. The amplified PCR products were subjected to agarose gel electrophoresis, and the single target DNA band was cut from the agarose gel and then recovered and purified using a DNA purification kit (Beijing Zhuangmeng Bio). The recovered target fragment was ligated to the vector at room temperature using a TA zero background rapid cloning kit (Beijing Zhuangmeng Bio). The ligation product was then transformed into DH5α Escherichia coli competent cells. After the transformation, the competent cells were added with 500 μL LB liquid culture medium and cultured at 37°C and 180 rpm for 1 h. Subsequently, 100 μL of the bacterial solution was spread on a LB solid plate containing 100 μg / mL ampicillin and cultured inverted at 37°C overnight. The positive clones on the plate were picked and PCR amplified using plasmid DNA primers M13F / M13R. The PCR reaction system was: single colony, upstream primer M13F (10µmol / L) 1μL, downstream primer M13R (10µmol / L) 1μL, DreamTaq Green PCR Master Mix (2×) 12.5μL, and ddH2O was added to 25μL. The PCR reaction conditions were: 94℃ pre-denaturation for 5min; 94℃ denaturation for 30s, 54℃ annealing for 30s, 72℃ extension for 90s, 30 cycles; 72℃ extension for 10min. The amplified PCR products were subjected to agarose gel electrophoresis, and the PCR products with the correct fragment size were sent to Guangzhou Tianyi Huiyuan Gene Technology Co., Ltd. for 16SrDNA full-length sequencing using M13F / M13R. The results are shown below.

[0048] 27F: 5′-AGAGTTTGATCCTGGCTCAG-3′;

[0049] 1492R: 5′-GGTTACCTTGTTACGACTT-3′;

[0050] M13F: 5′-TGTAAAACGACGGCCAGT-3′;

[0051] M13R: 5′-CAGGAAACAGCTATGACC-3′.

[0052] Sequencing results:

[0053]

[0054] (4) The sequencing results were compared on NCBI. The results showed that strain C03 and strain Annwoodia aquaesulis The 16S rDNA sequence of DSM 4255 had the highest similarity, with a sequence similarity of 94.97% and a sequence coverage of 99%. The 16S rDNA sequence of strain C03 was then compared with that of strain DSM 4255 using MEGA 11.0 software. Thiobacillaceae The 16S rDNA sequences of the strains of the family and the outgroup were aligned by ClustalW, and the maximum likelihood method was used. The Tamura-Nei model was selected, and the Ratesamong Sites parameter was selected as Gamma Distributed With Invariant Sites (G+I). The phylogenetic tree was constructed, and the topological structure of the phylogenetic tree was tested by 1000 bootstrap repeated samplings. The phylogenetic tree results are shown in Figure 2 According to the reference (Kook JK, et al. Genome-Based Reclassification of Fusobacterium nucleatum Subspecies at the Species Level. Curr Microbiol (2017) 74: 1137-1147.), the 16S rDNA sequence similarity of the strain is 98.65%, which can be used as the threshold for distinguishing the two strains. Therefore, based on the above 16S rDNA similarity, it can be judged that strain C03 is taxonomically similar to Annwoodia The existing species in the genus are different, so it can be judged that Annwoodia The resulting strain was named Annwoodia sp. C03, with the accession number GDMCC No: 65042, was deposited on August 28, 2024 at the Guangdong Microbial Culture Collection Center, Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou, Guangdong, China.

[0055] Example 2

[0056] Different oxygen concentrations Annwoodia The results of the influence of sulfur oxidation metabolism characteristics of sp. C03 are as follows:

[0057] Select the isolated, purified and identified AnnwoodiaA single colony of sp. C03 was inoculated in a serum bottle containing an enriched and screened inorganic culture medium (the composition is shown in Example 1), and nitrogen was pumped and added to obtain a headspace oxygen gas volume fraction of 3%. The culture temperature was 30°C, and the light-proof shaking (rotation speed was 150 rpm) was cultured for 3 days as a seed solution. A serum bottle containing an enriched and screened inorganic culture medium (wherein the concentration of Na2S2O3·5H2O was about 0.75 g / L (3 mmol / L)) was pumped and filled with nitrogen and oxygen was added to obtain a system with a headspace oxygen gas volume fraction of 0.1, 1, 3, 5, 8, 10, 15, and 21%, respectively. Then, the seed solution was inoculated into the serum bottle at an inoculum amount of 1% volume fraction, and the thiosulfate concentration, sulfate concentration, and pH were measured every 12 hours. The number of cells was counted at the 60th hour, and the total reaction time was 120 hours.

[0058] At different oxygen concentrations, Annwoodia The sulfur oxidation metabolism characteristics of sp. C03 in the culture medium containing sodium thiosulfate are as follows Figure 3 Shown: From Figure 3 As can be seen from A in the figure, in the culture medium inoculated with strain C03, the concentration of thiosulfate gradually decreased over time and basically entered a stable period after the 36th hour. In addition, the higher the oxygen concentration, the faster the thiosulfate consumption rate and the higher the conversion rate. In the range where the oxygen concentration is higher than 8%, all thiosulfate is consumed. Figure 3 As can be seen from B, in the culture solution inoculated with strain C03, the sulfate concentration gradually increased over time, and the amount generated basically reached stability at the 36th hour. In addition, the higher the oxygen concentration, the faster the sulfate generation rate and the higher the yield. Figure 3 As can be seen from C, the pH gradually decreases over time and gradually stabilizes after 36 hours. The higher the oxygen concentration, the lower the final pH of the culture medium. Figure 3 It can be seen from D that at the 60th hour of the reaction, when the oxygen concentration was 10%, the number of cells of strain C03 was the largest, and when the oxygen concentration was 0.1%, the number of cells of strain C03 was the smallest. Generally speaking, with the increase of oxygen concentration, the number of cells of strain C03 increased. However, since the sulfur oxidation rate in the experimental groups with oxygen concentrations of 15% and 21% was faster, the total conversion rate was higher, and the pH was lower, which affected the final activity of the bacteria, the number of cells decreased.

[0059] It can be seen that within the oxygen concentration range of 0.1% to 21%, the higher the oxygen concentration, the more conducive it is to sulfur oxidation by strain C03.

[0060] Example 3

[0061] Different oxygen concentrations Annwoodia The results of the influence of sp. C03 on colony formation on the enrichment screening solid inorganic medium plate are as follows:

[0062] The strain C03 seed solution prepared in Example 2 was diluted 1000 times, and 100 μL was inoculated onto an enrichment and screening solid inorganic culture medium plate (the composition is shown in Example 1) and evenly coated. There were 10 enrichment and screening solid inorganic culture medium plates in the experimental group, half of which were placed under conventional oxygen conditions (the volume fraction of headspace oxygen gas was about 21%) and cultured at 30°C, and the other half were placed under microaerobic conditions (the volume fraction of headspace oxygen gas was 5%) and cultured at 30°C to observe the formation of colonies on the enrichment and screening solid inorganic culture medium plates.

[0063] On the 5th day of culture, Figure 4 As shown in A, no colonies were observed on the enrichment screening solid inorganic medium plates cultured under normal oxygen conditions. Figure 4 In B, all enrichment screening solid inorganic medium plates cultured under microaerobic conditions have formed colonies visible to the naked eye. In subsequent observations, no colonies were observed on the enrichment screening solid inorganic medium plates cultured under normal oxygen conditions on the 12th, 30th, and 36th days of culture.

[0064] It can be seen that microaerobic conditions are more conducive to the formation of colonies by strain C03 on enrichment and screening solid inorganic culture medium plates, while conventional oxygen conditions with relatively high oxygen concentration are not conducive to the formation of colonies by strain C03 on enrichment and screening solid inorganic culture medium plates.

[0065] Example 4

[0066] Annwoodia The sulfur oxidation growth curve of sp. C03 under normal oxygen conditions is as follows:

[0067] Select the isolated, purified and identified Annwoodia A single colony of sp. C03 was inoculated in a serum bottle containing an enriched and screened inorganic culture medium (composition as shown in Example 1), with a headspace oxygen gas volume fraction of 21%, a culture temperature of 30°C, and cultured for 3 days in a dark and oscillating manner (speed of 150rpm) as a seed solution. A serum bottle containing an enriched and screened inorganic culture medium, with a headspace oxygen gas volume fraction of 21%, was inoculated with the prepared strain C03 seed solution in a serum bottle at an inoculum volume fraction of 1%, and the thiosulfate concentration, sulfate concentration, pH, and cell number were measured in phase for a certain period of time, and the total reaction time was 36h.

[0068] Annwoodia The sulfur oxidation growth curve of sp. C03 under normal oxygen conditions is as follows Figure 5 Shown: From Figure 5As can be seen from A, the concentration of thiosulfate ions in the culture medium of the inoculated strain C03 decreased rapidly over time and was completely consumed after 24 hours. The consumption rate of thiosulfate was 0.113mmol / L·h. Figure 5 As can be seen from B in the figure, the concentration of sulfate ions in the culture medium of the inoculated strain C03 gradually increased with time, reaching the maximum concentration after 24 hours and then tending to stabilize. The sulfate generation rate was 0.195 mmol / L·h, and the sulfate conversion rate reached 100%. Figure 5 It can be seen from C in that the pH of the culture medium of the inoculated strain C03 gradually decreased to about 5.83 over time and remained stable after 24 hours. Figure 5 As can be seen from Figure D, the cell number of strain C03 increased first and then decreased over time, reaching a maximum cell number of 1.41×10 at 27h. 8 CFU / mL.

[0069] Example 5

[0070] Annwoodia The results of sulfur oxidation and denitrification metabolic characteristics of sp. C03 under hypoxic conditions are as follows:

[0071] A serum bottle containing an enriched screening inorganic culture medium (wherein the concentration of Na2S2O3·5H2O is about 0.75 g / L (3 mmol / L), and 0.17 g / L (2 mmol / L) NaNO3 replaces NH4Cl as a nitrogen source and electron acceptor) is filled with nitrogen and the corresponding oxygen is added to obtain a system with a headspace oxygen gas volume fraction of 0.1%. Then, the seed solution prepared in Example 4 is inoculated into the serum bottle at an inoculum amount of 1% by volume, and the thiosulfate concentration, sulfate concentration, nitrate concentration, and nitrite concentration are phase-determined within a certain period of time, and the total reaction time is 204 h.

[0072] Annwoodia The growth curve of sulfur oxidation and denitrification of sp. C03 under hypoxic conditions is shown in Figure 6 Shown: From Figure 6 As can be seen from A, the concentration of thiosulfate ions in the culture medium of the inoculated strain C03 decreased rapidly over time. After 84 hours, the consumption rate of thiosulfate slowed down, and the total consumption rate of thiosulfate during the reaction was 0.0105mmol / L·h. Figure 6 As can be seen from B, the concentration of sulfate ions in the culture medium of the inoculated strain C03 increased rapidly over time, and the growth rate gradually slowed down after 84 hours, finally reaching a maximum concentration of 4.07mmol / L; Figure 6 As can be seen from C in the figure, the nitrate concentration in the culture medium of the inoculated strain C03 decreased rapidly and was almost completely consumed in 84 hours. The nitrate generation rate was 0.002 mmol / L·h. Figure 6As can be seen from D, there was almost no accumulation of nitrite in the culture medium of the inoculated strain C03, except for a nitrite concentration of 0.053 mmol / L detected at 84 h. The generation of N2O gas products was also detected in the headspace.

[0073] This shows that strain C03 can achieve sulfur oxidation and denitrification under low oxygen conditions.

[0074] Example 6

[0075] Annwoodia The results of the degradation characteristics of sp. C03 under normal oxygen conditions are as follows:

[0076] A serum bottle containing an enriched screening inorganic culture medium (wherein NaSCN replaces Na2S2O3·5H2O as a sulfur source with a concentration of about 0.082 g / L (2 mmol / L)) and a headspace oxygen concentration of 21% was inoculated with the seed solution prepared in Example 4 at an inoculum amount of 1% by volume into the serum bottle. After a certain period of time, the thiocyanate concentration and the sulfate concentration were measured. The total reaction time was 164 h.

[0077] Annwoodia The results of the degradation characteristics of sp. C03 under normal oxygen conditions are as follows Figure 7 As shown in the figure: the thiocyanate concentration in the culture medium of the inoculated strain C03 was completely consumed and converted into sulfate after 164 hours. The consumption rate of thiocyanate was 0.01 mmol / L·h, the removal rate reached 100%, and the generation rate of sulfate was 0.0135 mmol / L·h.

[0078] It can be seen that strain C03 can degrade thiocyanate under normal oxygen conditions.

[0079] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A chemoautotrophic plant Annwoodia A sulfur oxidizing bacteria strain characterized by: Chemoautotrophic Annwoodia The name of the sulfur oxidizing bacteria strain is Annwoodia sp. C03, with the accession number GDMCC No: 65042, was deposited on August 28, 2024 at the Guangdong Microbial Culture Collection Center, Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou, Guangdong, China.

2. The chemoautotrophic type according to claim 1 Annwoodia The invention relates to the use of a sulfur oxidizing bacteria strain, characterized in that: One of the following applications: (1) Application in redox sulfides; (2) Application in oxidation of reduced sulfides and simultaneous denitrification under hypoxic conditions; The redox sulfide is a reductive sulfide oxidized to sulfate; the reductive sulfide is at least one of elemental sulfur, thiosulfate, alkali metal sulfide, tetrathionate, thiocyanate and sulfite; The denitrification is to reduce nitrogen in an inorganic nitrogen source to N2O; the inorganic nitrogen source is at least one of nitrate and nitrite; The hypoxia is a condition where the oxygen concentration is 0.1% to 2%.

3. The use according to claim 2, characterized in that: The thiosulfate includes sodium thiosulfate, the alkali metal sulfide includes sodium sulfide, the tetrathionate includes sodium tetrathionate, the thiocyanate includes sodium thiocyanate, and the sulfite includes sodium sulfite; The nitrate includes sodium nitrate, and the nitrite includes sodium nitrite.

4. The use according to any one of claims 2 to 3, characterized in that: The application (1), The invention comprises the following steps: Annwoodia The sulfur-oxidizing bacteria strains were grown in an environment containing reduced sulfide; The application (2) comprises the following steps: Annwoodia The sulfur oxidizing bacteria strain is placed in a low oxygen environment containing reducing sulfide and inorganic nitrogen source for growth; the low oxygen refers to a condition where the oxygen concentration is 0.1% to 2%.

5. The use according to claim 4, characterized in that: The application (1), the growth is performed at an oxygen concentration of 0.1% to 21%.

6. The use according to claim 5, characterized in that: The application (1), the growth is performed under an oxygen concentration of 1% to 21%.

7. The use according to claim 4, characterized in that: In the application (1) or (2), the environment includes at least one of water and soil.

8. The use according to claim 4, characterized in that: In the application (1) or (2), the growth temperature is 25 to 35°C.

9. The use according to claim 8, characterized in that: In the application (1) or (2), the growth temperature is 30-35°C.

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