Alkalophilic halomonas with efficient sulfur oxidation function as well as culture method and application thereof

Abstract

The invention relates to a halomonas alkali with an efficient sulfur oxidation function, a culture method and application of the halomonas alkali, the halomonas alkali is named as DS-4, the preservation number of the halomonas alkali is CGMCC (China General Microbiological Culture Collection Center) No.: 30669, and the preservation date of the halomonas alkali is May 17, 2024. According to the present invention, the optimal desulfurization conditions of the halomonas alkalicola DS-4, the sulfide removal characteristics of the halomonas alkalicola DS-4, and the product of the halomonas alkalicola DS-4 are researched; the optimal desulfurization condition of the strain is that the temperature is 35 DEG C, the pH value is 8-9, the rotating speed of a shaking table is 180rpm, and the inoculation amount is controlled so that the culture medium OD600 is 0.08. Under the optimum condition, the removal effect on 1200mg / L sulfide is up to 99.93% after the strain is cultured for 11h. According to removal and main metabolite analysis of 400 mg / L sulfide, the removal rate of S < 2-> reaches 99.9% within 5 h, the main product is elemental sulfur, and the yield of the elemental sulfur reaches up to 75.55%. The strain has a wide application prospect in the aspects of treatment of hydrogen sulfide-containing gas and sulfide-containing wastewater and sulfur resource recovery.

Description

Technical Field

[0001] The invention belongs to the technical field of environmental biological application, and relates to an alkaliphilic halomonas with high-efficiency sulfur oxidation function, a cultivation method and application thereof. Background Art

[0002] Energy and environmental crises are increasingly becoming a problem that plagues human sustainable development, and people's demand for renewable energy and environmental governance is becoming more and more urgent. The demand for low-carbon energy such as biogas and natural gas is increasing year by year. The hydrogen sulfide contained in them will not only corrode pipelines and equipment, but also cause serious odors that affect the environment and even seriously endanger human health; the rapid development of industries such as wastewater treatment, papermaking, animal husbandry, and metallurgical industry has brought a large amount of gaseous sulfur-containing odorous gases and sulfur-rich sulfide wastewater, which poses a serious threat to the water ecological environment. Accelerating the removal of hydrogen sulfide gas in energy sources such as biogas and natural gas and the treatment of sulfide-rich wastewater has become an environmental problem that my country urgently needs to solve.

[0003] Hydrogen sulfide (H2S) is a toxic and smelly gas produced in nature and during human activities. Its olfactory threshold is only 0.0041ppm. The presence of trace amounts of H2S in the air (>2ppm) will cause damage to human health. If the concentration rises to 100ppm, it will paralyze the human olfactory nerves, and exceeding 400ppm may even cause death. Its aqueous solution is sulfide or hydrosulfide, which can easily cause black and smelly water bodies and has strong biological toxicity; when exposed to the atmosphere, it is easily oxidized to SO2 or SO3, and then forms acid rain. It can be seen that the presence of sulfides will have adverse effects on people, the environment and society.

[0004] At present, there are many industrially effective methods for treating malodorous sulfide wastewater and hydrogen sulfide gas at home and abroad. According to their weak acidity and strong reducing properties, they can be divided into physical, chemical and biological methods. Among them, biological desulfurization has gradually become the research focus and hotspot in the current desulfurization industry due to its advantages such as simple equipment, mild reaction conditions, low cost, high efficiency, no need for catalysts, and no secondary pollution. The key technology of biological sulfur oxidation is the screening of efficient sulfur oxidizing bacteria (SOB), using the enzymes it contains to catalyze the desulfurization of sulfur-containing compounds (H2S, S 2-) is converted into elemental sulfur or sulfate. There are currently two technologies for biological removal of hydrogen sulfide from biogas, natural gas or other gases that have been industrially applied: the Bio-SR process and the Shell-Paques process. The Bio-SR process uses an aqueous solution of ferric sulfate as an absorbent to absorb and oxidize hydrogen sulfide. At the same time, ferric sulfate is reduced to ferrous sulfate, and ferrous sulfate is oxidized by acidophilic bacteria such as Thiobacillus ferrooxidans, thereby realizing the recycling of iron. However, the Bio-SR process has harsh operating conditions and needs to operate under strongly acidic conditions. It has extremely high requirements for the corrosion resistance of the equipment, and the produced potassium ferrosite by-product is easy to clog the equipment. Although the Bio-SR process was developed and applied in the 1980s, its promotion speed has been slow (Anyang, East China University of Science and Technology, 2021.). The Shell-Paques process uses sodium carbonate aqueous solution (containing sulfur-containing bacteria brought in by the circulating fluid and the nutrient solution required for their growth) as an absorbent, with the pH controlled at 8-10. Under the action of a mixed bacteria composed of various thiobacilli, sulfide is directly oxidized into elemental sulfur, which overflows from the top of the bioreactor in the form of slurry. Since biosulfur is hydrophilic and has good fluidity, sulfur blockage and equipment corrosion can be avoided (Gao Shuang, Harbin Institute of Technology, 2021; Liu Fei, Environmental Science and Management, 2018, 43(11):76-79.). The Shell-Paques process has the advantages of simple operation, low energy consumption, internal circulation of alkali solution, and automatic regeneration of strains (Ma Ning, Petroleum and Natural Gas Industry, 2019, 40(9):47-50.), and is widely used, but the screening and cultivation of new and efficient desulfurization strains remains the focus and difficulty in this field.

[0005] At present, most of the biological oxidative desulfurization strains reported in the literature are chemoautotrophic or heterotrophic desulfurization, and their living environment is mainly a neutral pH environment. The advantage of chemoautotrophic desulfurization strains is that they do not require exogenous organic nutrients, and are currently widely used in biological desulfurization, but these strains grow slowly, which will slow down the start-up of the desulfurization project and affect the desulfurization efficiency of the entire process. For example, Juan P. et al. used autotrophic desulfurization strains to start the desulfurization project. Initially, the abundance of each bacteria in the community was less than 10%. After 105 days of cultivation, the peak degree of each strain reached more than 20% (Juan PM et al, Chemosphere, 2010, 80 (8): 872-880; Zhao Peng, University of Chinese Academy of Sciences, 2017.) Heterotrophic desulfurization bacteria are currently mostly used for the removal of organic sulfides (Wei, Z Set al, Environmental Science, 2015.). They have the characteristics of rapid growth and high efficiency in removing organic sulfides, but there are few reports on the removal of inorganic sulfides by organic heterotrophic desulfurization bacteria. Due to the pH tolerance of neutral sulfur oxidizing bacteria, the operating pH of the desulfurization process cannot exceed 8.5, and the salinity cannot exceed 0.5 mol / L Na + (Mu Tingzhen et al., Journal of Biotechnology, 2021, 37(02): 461-472.), under this process condition, the system's ability to absorb H2S is very limited, and its impact resistance is weak. Based on the above characteristics, autotrophic bacteria and desulfurization bacteria grown under neutral pH conditions are subject to certain limitations in practical applications. Therefore, the screening and application of alkaliphilic heterotrophic sulfur-oxidizing bacteria have received great attention. For example, Wageningen University and Delft University of Technology in the Netherlands have jointly developed a laboratory-scale halophilic and alkaliphilic biological desulfurization system, demonstrating the application potential of Thioalkalivibrio in biological desulfurization (Sorokin DY et al, Applied Microbiology and Biotechnology, 2008, 80(6): 965-975; Van den BPL F et al, Biotechnology and Bioengineering, 2007, 97(5): 1053-1063.); The strain DS-8 screened by Zhao Peng grew in heterotrophic medium for 28 hours and its OD 600 The value increased from 0.135 to 1.358 (Zhao Peng, University of Chinese Academy of Sciences, 2017.). Developing alkaliphilic heterotrophic sulfur-oxidizing bacteria resources and using them for the treatment of hydrogen sulfide gas and sulfide wastewater can recover elemental sulfur resources while treating sulfur pollution, which has a very broad application prospect. Summary of the invention

[0006] The present invention aims to solve the problems of poor alkali resistance, slow growth rate and long sulfide oxidation cycle of autotrophic or heterotrophic sulfur oxidizing bacteria grown under the above neutral pH conditions, and provides a strain of alkaliphilic halomonas with efficient sulfur oxidation function and its application. The sulfur oxidizing bacteria DS-4 provided by the present invention has fast growth rate, wide adaptability and good salt tolerance. + Under the condition of 1M concentration, the removal efficiency of 1200mg / L sulfide is as high as 99.93% after 11h of cultivation, and the removal efficiency of 400mg / L sulfide is as high as 99.9% after 5h of cultivation. The main product of sulfur oxidation is elemental sulfur.

[0007] The technical solution of the present invention:

[0008] A strain of alkaliphilic halomonas with efficient sulfur oxidation function, characterized in that the bacterium is alkaliphilic halomonas Halomonas alkalicola DS-4, the preservation number is CGMCC No.30669, the preservation location is: General Microbiology Center of China Microbiological Culture Collection Administration, and the preservation time is: May 17, 2024.

[0009] The culturing method of the alkaliphilic halomonas comprises the following steps:

[0010] The sludge used for separating and screening strains in the present invention comes from the bottom mud of Suminjilin Lake in Badain Jaran Desert, Alxa Right Banner, Alxa League, Inner Mongolia. It is collected at 0.5-1m underwater with a mud bucket, and stones and animal and plant remains are removed. Under aerobic conditions, the strain alkaliphilic halomonas is inoculated in LB medium, pH=9, and cultured in a constant temperature shaker at 30°C and 150rpm;

[0011] LB medium components: peptone 10g / L, NaCl 10g / L, yeast extract powder 5g / L.

[0012] The application of the alkaliphilic halomonas refers to the alkaliphilic halomonas having a highly efficient sulfur oxidation function.

[0013] The specific applications are as follows:

[0014] The alkaliphilic Halomonas bacteria were inoculated into LB medium and cultured to the end of the logarithmic phase, centrifuged at 10,000 rpm for 5 min, the supernatant was discarded, the bacterial cake was washed twice with saline, and finally resuspended with saline to obtain a bacterial suspension. With a certain inoculum amount and under certain conditions, the bacterial suspension was added to the sulfide medium to remove sulfide in hydrogen sulfide gas or sulfide in wastewater and produce elemental sulfur;

[0015] The inoculation amount is the OD value after the bacterial suspension is added to the sulfide removal medium. 600 The absorbance is expressed as 600is 0.02-0.10, sulfide removal medium components: C4H4Na2O4 0.5-2g / L, NaHCO3 5-10g / L, NH4Cl 0.5-1.5g / L, MgCl2 0.2-0.5g / L, K2HPO4·3H2O 0.2-1g / L, NaCl 0-90g / L, sulfide concentration is 0.2-2.5g / L, use 1M HCl and 1M NaOH to adjust pH.

[0016] The optimal desulfurization conditions for the DS-4 strain are: 35°C, pH = 8-9, shaking speed of 180 rpm, and controlling the inoculation amount to make the culture medium OD 600 is 0.08.

[0017] The DS-4 strain has a removal rate of up to 99.93% for 1200 mg / L sulfide after being cultured for 11 hours, a removal rate of up to 99.9% for 400 mg / L sulfide after being cultured for 5 hours, and a yield of elemental sulfur of up to 75.55%.

[0018] Beneficial effects of the present invention:

[0019] (1) The alkaliphilic Halomonas alkalicola DS-4 provided by the present invention is a salt-tolerant, heterotrophic sulfur-oxidizing strain screened from a saline-alkali lake sediment sample from the Badain Jaran Desert in Alxa Right Banner, Alxa League, Inner Mongolia. It can + =0.1~2M conditions and remove sulfide. In LB medium, the bacteria enters the logarithmic growth phase in 8 hours, with a fast growth rate. Under the optimal conditions, the bacteria achieves a 99.93% removal effect on 1200mg / L sulfide in 11 hours, which is better than the existing experimental results. The removal experiment of 400mg / L sulfide was carried out, and the analysis showed that the product was mainly elemental sulfur, and the elemental sulfur yield was as high as 75.55%. The strain has broad application prospects in the treatment of hydrogen sulfide gas and sulfide wastewater.

[0020] (2) The alkaliphilic Halomonas alkalicola DS-4 provided by the present invention, in the application process of treating sulfide-containing gas and sulfide wastewater, under microaerobic conditions, the strain oxidizes sulfide, and the product is mainly elemental sulfur, and the elemental sulfur is secreted outside the cell and no longer oxidized, which effectively prevents the secondary pollution of water bodies and solution acidification caused by sulfate formation. The generated elemental sulfur can be used in the fields of sulfur production, agriculture and medicine, which not only solves the pollution but also recovers sulfur resources, generating environmental benefits while also having certain economic benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 (a) is a colony morphology of strain DS-4 on a plate culture medium;

[0022] Figure 1(b) is a scanning electron micrograph of strain DS-4 on a plate culture medium;

[0023] Figure 2 Phylogenetic tree of strain DS-4;

[0024] Figure 3(a) shows the growth of strain DS-4 under different inoculation conditions and the removal effect of sulfide;

[0025] FIG3( b ) is a graph showing the growth of strain DS-4 under different temperature conditions and the removal effect on sulfide;

[0026] Figure 3(c) shows the growth of strain DS-4 under different pH conditions and the removal effect on sulfide;

[0027] FIG3( d ) shows the growth of strain DS-4 under different shaking speed conditions and the removal effect of sulfide;

[0028] Figure 4 This is the removal effect of strain DS-4 on different concentrations of sulfide under optimal conditions;

[0029] FIG5 is a comparison of the sulfide removal efficiency of strain DS-4 before and after condition optimization;

[0030] Figure 6 is a graph showing sulfide removal and product generation by strain DS-4; wherein (a) is a graph showing the relationship between sulfide removal and product generation, and (b) is a graph showing the relationship between sulfide removal rate and product generation rate. DETAILED DESCRIPTION

[0031] To further illustrate the technical means and effects of the present invention, the present invention is further described below in combination with the technical scheme, drawings and application examples. It is understood that the specific implementation methods described herein are only used to explain the present invention, rather than to limit the present invention.

[0032] Example 1: Screening of strains

[0033] (1) Enrichment and separation and purification

[0034] Take 5g of sediment and add it to 100mL of enrichment medium. Incubate the mixture in a shaker at 30℃ and 150rpm for enrichment. Take samples at regular intervals to measure S 2- concentration, wait for S in the system 2- After the removal effect is stable, take 10mL of enriched bacterial solution and add it to fresh enriched culture medium. Continue to culture according to the above method to continuously improve S 2-The enrichment process was completed after 10 times of continuous enrichment culture. Enrichment medium composition: C4H4Na2O4 1.5g / L, NaHCO3 5g / L, NH4Cl 1g / L, MgCl2 0.2g / L, K2HPO4·3H2O 0.5g / L, NaCl 2.5g / L, sterilized at 115℃ for 30min. After cooling, add Na2S·9H2O solution filtered through 0.22μm filter membrane to make the solution S 2- The concentrations were 0.2, 0.4, and 0.6 g / L, respectively. 1 M HCl and 1 M NaOH were used to adjust the pH to 9.

[0035] Take 100 μL of the enriched bacterial solution and dilute it with physiological saline to 10% of the original bacterial solution. -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 times, take 50 μL of each and spread on sulfide screening medium and culture for 24-48 hours until strains with different morphologies grow, and use LB medium for activation. Sulfide screening medium composition: peptone 10g / L, NaCl 10g / L, yeast extract 5g / L, agar 20g / L, pH = 9. Sterilize at 121℃ for 20min. After cooling slightly, add Na2S·9H2O solution filtered through 0.22μm filter membrane to make the medium S 2- The concentration is 0.2g / L.

[0036] (2) Initial screening

[0037] Preparation method of bacterial suspension: inoculate the strain into LB medium and culture it to the end of the logarithmic phase, centrifuge at 10000rpm for 5min, pour off the supernatant, wash the bacterial cake twice with saline, and finally resuspend it with saline.

[0038] A certain amount of bacterial suspension was added to the primary screening culture medium to make the OD of the culture medium 600 =0.1, and the initial screening was carried out according to the sulfide removal effect. The mixture was cultured at 30°C and 150 rpm for 6 h, and samples were taken every 1 h to measure S 2- , finally select S 2- The strains with better removal effect enter the rescreening. The components of the primary screening medium: C4H4Na2O4 1.5g / L, NaHCO3 5g / L, NH4Cl 1g / L, MgCl2 0.2g / L, K2HPO4·3H2O 0.5g / L, NaCl 2.5g / L, sterilized at 115℃ for 30min. After cooling, add 0.2g / L Na2S·9H2O filtered through a 0.22μm filter membrane. Use 1M HCl and 1M NaOH to adjust pH to 9.

[0039] (3) Rescreening

[0040] A certain amount of bacterial suspension was added to the rescreening medium to make the OD of the medium 600 =0.1, incubate at 30°C, 150 rpm for 6 h, take samples and measure S 2- Concentration to determine the sulfide removal effect. Rescreening medium composition: C4H4Na2O4 1.5g / L, NaHCO3 5g / L, NH4Cl 1g / L, MgCl2 0.2g / L, K2HPO4·3H2O 0.5g / L, NaCl 2.5g / L, sterilized at 115℃ for 30min. After cooling, add 0.4g / L Na2S·9H2O filtered through a 0.22μm filter membrane. Use 1M HCl and 1MNaOH for adjustment.

[0041] (4) Identification of strains

[0042] The strain DS-4 was identified by PCR amplification using primers (27F-1492R).

[0043] The 16S rDNA sequence table of the alkaliphilic Halomonas alkalicola DS-4 is SEQ ID No. 1, as shown below:

[0044]

[0045] Application Example 1: Application of a strain of alkaliphilic Halomonas DS-4 with efficient sulfur oxidation function in sulfide removal

[0046] The strain DS-4 was inoculated into LB medium and cultured to the end of the logarithmic phase, centrifuged at 10000rpm for 5min, the supernatant was discarded, the bacterial cake was washed twice with saline, and finally resuspended with saline to prepare a bacterial suspension. With a certain inoculum amount and under certain conditions, the bacterial suspension was added to the sulfide medium for the strain removal sulfide experiment.

[0047] The sulfide removal experiment was carried out in a 250 mL conical flask filled with 100 mL of culture medium and covered with a sealing film. Three experimental groups and three control groups were set up for each experiment.

[0048] The specific experiments are as follows:

[0049] (1) Effect of inoculum dose on strain growth and S 2- The removal effect

[0050] Effects of different inoculum doses on the growth of strains and S 2- The removal effect is shown in Figure 3(a). The inoculum amount is the OD value after the bacterial suspension is added to the sulfide removal medium. 600 As shown in Figure 3(a), the growth of the strain (expressed as protein concentration, the same below) and the effect of S 2- The removal efficiency of the bacteria increases with the increase of the inoculum amount. 600 0.02-0.08, S 2- The removal rate showed a rapid growth trend. 2- The removal rate was 31.2%-91.1%; when the inoculum volume was controlled to make the culture medium OD 600 When S is 0.08-0.10, 2- The removal rate growth slowed down, and the S 2- The removal rate was 91.4%. 600 When it is greater than or equal to 0.08, the hysteresis period of microbial growth can be reduced, thereby quickly removing S 2- However, if the inoculum amount is too large, it may cause competition for nutrients between bacteria, thus slowing down the removal effect. Therefore, the inoculum amount should be controlled to make the culture medium OD 600 0.08-0.10 is most suitable.

[0051] (2) Effects of temperature on the growth of strains and S 2- The removal effect

[0052] Temperature is one of the important factors affecting enzyme activity in microorganisms. Different temperatures have an impact on the growth of strains and the activity of S2- The removal effect of is shown in Figure 3(b). As shown in Figure 3(b), when the temperature rises from 20℃ to 40℃, the growth of the strain and S 2- The removal efficiency of S 2- The removal rate of S was the highest, and the removal rate of S 2- The removal rate was 90.5%. When the temperature increased from 20℃ to 35℃, the S 2- The removal rate gradually increased from 21.0% to 90.5%. When the temperature increased from 35℃ to 40℃, the S 2- The removal rate dropped from 90.5% to 86.8%. The temperature range of strain DS-4 was wide, and its optimum growth and 2- The removal temperature was 35°C.

[0053] (3) Effect of initial pH on strain growth and S 2- The removal effect

[0054] pH is one of the important factors affecting the growth of microorganisms. Different initial pH has an impact on the growth of strains and the effect of S 2- The removal effect of is shown in Figure 3(c). As shown in Figure 3(c), S 2- The removal rate showed a slow downward trend with the increase of initial pH. When the initial pH value was between 7 and 8, the growth of the strain was good. 2- The removal rate of S was higher, and the removal rate of S 2- The removal rate reached about 90.0%. When pH = 9, the growth of the strain and S 2- The removal rate of S was also high. 2- The removal rate was 82.8%. When the pH continued to rise, the growth of the strain and S 2- The above results show that the strain DS-4 has a wide pH adaptability range and a strong sulfide removal ability in a neutral to alkaline environment. In order to reduce H2S escape and increase its absorption performance, it is more appropriate to adjust pH to 9.

[0055] (4) Effect of shaker speed on strain growth and S 2- The removal effect

[0056] The shaking speed leads to different dissolved oxygen content in the solution, which in turn affects the growth of the strain and its effect on S 2- Effect of different shaking speeds on the growth of strains and the removal of S 2- The removal effect of S is shown in Figure 3(d). As shown in Figure 3(d), the growth of strain DS-4 at different shaking speeds and its effect on S 2- When the shaker speed was 60-180 rpm, as the shaker speed increased, the growth of the strain and its removal effect on S 2-When the speed was between 180-210 rpm, with the increase of the shaking speed, the growth of the strain and the removal rate of S 2- The removal rate of DS-4 began to decline slowly. This indicates that the growth rate of strain DS-4 at 180 rpm is similar to that of S. 2- Optimal removal effect.

[0057] (5) The strain DS-4 was sensitive to different concentrations of S under optimal conditions. 2- The removal effect

[0058] The prepared bacterial solution was inoculated into an enrichment medium containing different concentrations of sulfide (200, 400, 600, 800, 1000, 1200 mg / L), and the pH of the medium was adjusted to 9 with HCl and NaOH solutions filtered through a 0.22 μm sterilized microporous filter membrane. The medium was cultured at 35°C and 180 rpm, and the S in the reaction solution was measured every 1 h. 2- Content. Different concentrations of S 2- The strain S under the conditions 2- Removal effect Figure 4 The test results showed that strain DS-4 was resistant to 200, 400, 600, 800, 1000, and 1200 mg / L S 2- The removal rates were all above 99.9%, and the removal rates were fast, namely 4, 5, 6, 8, 10, and 11 h, respectively, indicating the high efficiency of the strain in removing sulfide.

[0059] (6) Effect of strain DS-4 on S before and after optimization 2- Comparison of removal effects

[0060] The present invention explores the effect of strain DS-4 on S 2- The removal effect of S after optimization is compared with that before optimization. The results are shown in Figure 5(a) and Figure 5(b). At the same time, the optimized S 2- The removal rate is higher than before optimization, at 400mg / LS 2- Under the conditions of concentration, the removal rate reached 92.9% after 4 hours of optimization, which was 38.6% higher than that before optimization. This showed that under the reaction conditions of initial pH = 9, 35 °C, and 180 rpm, the strain DS-4 could remove S 2- The ability has been significantly improved.

[0061] (7) Sulfide removal and main metabolites of strain DS-4

[0062] Under the optimal conditions (control the inoculum amount so that the culture medium OD 600 0.08, temperature 35°C, initial pH = 9, shaker speed 180 rpm), strain DS-4 removed S 2-The changes in the sulfur form during the process are shown in Figure 6(a) and Figure 6(b). 2- During the biological removal process, different forms of inorganic sulfur were detected, including S 2- , S 0 、S2O3 2- 、SO3 2- and SO4 2- The changes in the concentration of these inorganic sulfurs show obvious differences throughout the reaction process. 2- As an electron donor, it is oxidized to other inorganic sulfur states. Its concentration drops rapidly in the first 4 hours and then tends to level off. After 5 hours of reaction, the concentration of S in the water sample is 2- The residual concentration of S2O3 is 0.66mg / L, and the oxidation rate reaches 99.9%. 2- The concentration of SO4 increases first and then decreases as the reaction proceeds, indicating that as an intermediate in the formation of elemental sulfur, there are two processes of generation and consumption during the reaction; 2- and SO3 2- The concentration was very low during the entire reaction process, and the elemental sulfur content showed a trend of increasing as the reaction proceeded. When the sulfide was completely removed, the elemental sulfur yield was 75.55%, indicating that elemental sulfur was S 2- The main product of biological removal process.

Claims

1. An alkaliphilic Halomonas strain with efficient sulfur oxidation function, It is characterized by, The bacterium is Halomonas alkalicola DS-4, and the preservation number is CGMCC No. 30669. The preservation location: General Microbiology Center of the China Microbial Culture Collection Committee, and the preservation time: May 17, 2024.

2. A strain of Halomonas alkalophilus with efficient sulfur oxidation function according to claim 1, It is characterized by, The 16S rDNA sequence list SEQ ID of the Halomonas alkalicola DS-4 No.1: GGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGCAGCACGGGAAGCTTGCTTCCTGGTGGCGAGCGGCGGACGGGTGAGTAATGCATAGAAATCTGCCCGGTAGTGGGGGATAACCTGGGGAAACCCAGGCTAATACCGCATACGTCCTACGGGAGAAAGCAGG GGATCTTCGGACCTTGCGCTATCGGATGAGTCTATGTCGGATTAGCTTGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCCGTAGCTGGTCTGAGAGGATGATCAGCCACATCGGGACTGAGACACGGCCCGAACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCT GATCCAGCCATGCCGCGTGTGTGAAGAAGGCCTTCGGGGTTGTAAAGCACTTTCAGTGGGGAAGAAAGCCTTGGGGCTAATACCCTCGAGGAAGGACATCACCCACAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCGAGCGTTAATCGGAATTACTGGGCGTA AAGCGCGCGTAGGCGGTCTGATAAGCCGGTTGTGAAAGCCCCGGGCTCAACCTGGGAACGGCATCCGGAACTGTCAGGCTAGAGTGCAGGAGAGGAAGGTAGAATTCCCGGTGTAGCGGTGAAATGCGTAGAGATCGGGAGGAATACCAGTGGCGAAGGCGGCCTTCTGGACTGACACTGACGCT GAGGTGCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTAGCCGTTGGGCTCCTTGAGAGCTTTGTGGCGCAGTTAACGCGATAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGG TGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTACCCTTGACATCGTGCGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACGCACAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTATTTG CCAGCGAGTAATGTCGGGAACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAAGTCATCATGGCCCTTACGGGTAGGGCTACACACGTGCTACAATGGCCGGTACAAAGGGTTGCGAAGCCGCGAGGTGGAGCTAATCCCGAAAAGCTGGTCTCAGTCCGGATCGGA GTCTGCAACTCGACTCCGTGAAGTCGGAATCGCTAGTAATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGACTGCACCAGAAGTGGTTAGCCTAACTTCGGAGGGCGATCACCACGGTGTGGTTCATGACTGGGGTGAAGTCGT.

3. The culture method of Halomonas alkalophilus according to claim 1 or 2, It is characterized by, The steps are as follows: Under aerobic conditions, inoculate the strain Halomonas alkalophilus into LB culture medium, pH=9, and culture it on a constant temperature shaker at 30°C at 150 rpm; LB medium components: peptone 10g / L, NaCl 10g / L, yeast extract 5g / L.

4. Application of Halomonas alkalophilus according to claim 1 or 2, Its characteristics are: Alkalophilic Halomonas has efficient sulfur oxidation function.

5. Application of Halomonas alkaliphila according to claim 4, Its characteristics are: Inoculate Halomonas alkalophilus into LB culture medium and culture it to the end of the logarithmic phase, centrifuge at 10,000 rpm for 5 minutes, discard the supernatant, wash the bacterial cake twice with physiological saline, and finally resuspend with physiological saline to prepare a bacterial suspension; Add the bacterial suspension to the sulfide culture medium, and use the bacterial strain to remove the alkali, absorb the sulfide in the hydrogen sulfide gas or the sulfide in the wastewater, and produce elemental sulfur.

6. Application of Halomonas alkalophilus according to claim 5, It is characterized by: among them, The inoculation amount is based on the bacterial suspension added to the sulfide removal medium and the OD 600 The absorbance indicates that the amount of inoculated bacteria is controlled to make the culture medium OD 600 0.02-0.10, sulfide removal medium components: C 4 H 4 Na 2 O 4 0.5~2g / L, NaHCO 3 5~10g / L, NH 4 Cl 0.5~1.5g / L, MgCl 2 0.2~0.5g / L,K 2 HPO 4 ·3H 2 O 0.2~1g / L, NaCl 0~90g / L, sulfide concentration 0.2-2.5g / L, use 1M HCl and 1M NaOH to adjust the pH.

7. Application according to claim 4 or 5 or 6, Its characteristics are: The optimal desulfurization conditions for the DS-4 strain are: 35°C, pH=8-9, shaking table speed of 180rpm, and controlling the amount of bacteria to make the culture medium OD 600 is 0.

08.

8. Application according to claim 7, Its characteristics are: The DS-4 strain has a removal rate of 1200 mg / L sulfide as high as 99.93% after 11 hours of cultivation, a removal rate of 400 mg / L sulfide as high as 99.9% after 5 hours of cultivation, and the elemental sulfur yield is as high as 75.55%.

Images