Staphylococcus saprophyticus NCSL-YWH2 with lithium ion enrichment capacity and application of staphylococcus saprophyticus NCSL-YWH2
By culturing and enriching Staphylococcus saprophyticus NCSL-YWH2, the problems of high cost and environmental unfriendliness of existing brine lithium extraction technology have been solved, realizing efficient, green and low-carbon lithium resource recovery, which is suitable for lithium ion enrichment in various lithium-containing water bodies.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-31
- Publication Date
- 2026-03-31
AI Technical Summary
Existing brine lithium extraction technologies are costly, environmentally unfriendly, and difficult to apply to low-concentration lithium-ion systems. There is a lack of green and environmentally friendly biological lithium extraction technologies.
Using a strain of Staphylococcus saprophyticus NCSL-YWH2, lithium ions were efficiently enriched in various lithium-containing water bodies, including salt lake brine, geothermal hot spring water, oil and gas field produced water, and seawater, through cultivation and enrichment methods. Inorganic, organic, and composite carriers were used for lithium enrichment.
It achieves efficient, green, and low-carbon lithium resource recovery, with a removal rate of up to 95% and an enrichment of up to 120 mg/g of stem cells. It is suitable for different types of water bodies, including low-concentration mineral water and high-concentration industrial wastewater, and has broad practical application potential.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of biological resource recycling technology, specifically, it relates to a strain of Staphylococcus saprophyticus NCSL-YWH2 with lithium ion enrichment capacity and its application. Background Technology
[0003] The technologies that have received widespread attention in lithium extraction from brine include precipitation, adsorption, membrane separation, solvent extraction, and electrochemical methods, while green and environmentally friendly biological lithium extraction technologies are rarely reported.
[0004] In recent years, yeast has been used as a model microorganism to study the channel proteins for lithium ion entry into cells, confirming the ability of microorganisms to take up lithium ions. However, there are few reports in literature or patents on the utilization of microorganisms to enrich lithium ions in liquids and thus realize lithium resource utilization. Currently, there are reports on Staphylococcus species that can tolerate high-concentration lithium environments, but there are no reports on their enrichment of lithium ions in water. Summary of the Invention
[0005] The purpose of this invention is to overcome the problems of high cost, environmental unfriendliness, and difficulty in application to low-concentration lithium ion systems in existing brine lithium extraction technologies, and to develop a green and environmentally friendly biological lithium extraction technology. This invention provides a strain of Staphylococcus saprophyticus NCSL-YWH2 with highly efficient lithium ion enrichment capacity and its application method for extracting lithium resources in various lithium-containing water bodies.
[0006] To achieve the above objectives, the first aspect of the present invention provides a strain of Staphylococcus saprophyticus NCSL-YWH2, with accession number CGMCC No. 35847.
[0007] Furthermore, the nucleotide sequence of the 16S rDNA of the *Staphylococcus saprophyticus* NCSL-YWH2 is shown in SEQ ID NO.1.
[0008] A second aspect of the present invention provides a method for culturing the *Staphylococcus saprophyticus* NCSL-YWH2.
[0009] Preferably, the method for culturing the *Staphylococcus saprophyticus* NCSL-YWH2 includes:
[0010] Activation culture: The strain was inoculated into LB liquid medium for activation culture at a temperature of 10-35℃ for 1-3 days and a shaking speed of 100-250 rpm until the logarithmic growth phase was reached.
[0011] Expanded culture: The activated strain was inoculated into liquid culture medium at an inoculation rate of 10% (v / v) for expanded culture. The culture temperature was 10-35℃, the culture time was 1-10 days, and the shaking speed was 100-250 rpm.
[0012] Furthermore, the LB liquid culture medium used in the activation culture consists of: 10 g / L tryptone, 5 g / L yeast extract, and 10 g / L sodium chloride.
[0013] Furthermore, the liquid culture medium used in the expanded culture consists of: carbon source 1–100 g / L, ammonium chloride 0.2–20 g / L, potassium dihydrogen phosphate 0.05–8 g / L, magnesium sulfate heptahydrate 0.2–5 g / L, and lithium chloride 0–100 g / L.
[0014] The carbon source is selected from one or more combinations of glucose, starch, sucrose, glycerol, and molasses.
[0015] A third aspect of the invention provides the application of the *Staphylococcus saprophyticus* NCSL-YWH2 strain for enriching lithium in lithium-containing liquids.
[0016] Preferably, the lithium element enriched in the lithium-containing liquid is obtained by directly using the cells of Staphylococcus saprophyticus NCSL-YWH2, attaching them to the surface of other functional carriers, or preparing them into encapsulated particles.
[0017] According to the present invention, the functional carrier includes inorganic carriers, organic polymer carriers, and composite carriers;
[0018] According to the present invention, the materials of the encapsulated particles include natural polymer materials, synthetic polymer materials, and composite materials.
[0019] Furthermore, the lithium ion concentration of the lithium-containing liquid ranges from 0.2 to 2000 mg / L.
[0020] According to the present invention, the lithium-containing liquid is selected from one or more of the following: salt lake brine, geothermal hot springs and hot brine, produced water from oil and gas fields, lithium-containing mineral water, seawater, lithium ore leachate, lithium battery production wastewater, wastewater from the recycling process of retired lithium-ion batteries, wastewater from the industrial production and recycling processes of lithium-containing chemicals and materials such as glass ceramics, lubricants, alloys, and pharmaceuticals, simulated lithium-containing liquid, and liquids generated during the processing of the above-mentioned lithium-containing liquids.
[0021] The present invention has the following beneficial effects:
[0022] 1. Staphylococcus saprophyticus NCSL-YWH2 can effectively enrich lithium ions in various lithium-containing water bodies (such as salt lake brine, geothermal hot spring water, oil and gas field produced water, seawater, industrial wastewater, etc.), with a removal rate of up to 95% and an enrichment amount of up to 120 mg / g stem cells, resulting in high lithium resource recovery efficiency.
[0023] 2. Staphylococcus saprophyticus NCSL-YWH2 has a wide tolerance range for lithium ion concentration (0-100 g / L lithium chloride) and can be applied to different types of water bodies, including low-concentration mineral water and high-concentration industrial wastewater. It has strong practical application potential and promotion value.
[0024] 3. This invention uses a microbial method to extract lithium, which avoids the problems of large reagent consumption and high energy consumption of traditional chemical methods. It has the characteristics of being green, low-carbon, and renewable, and meets the requirements of sustainable development. Attached Figure Description
[0025] Figure 1 The morphology of Staphylococcus saprophyticus NCSL-YWH2 on LB agar plates is shown.
[0026] Figure 2 Phylogenetic tree of Staphylococcus saprophyticus NCSL-YWH2 strain.
[0027] Preservation Matters
[0028] The lithium-rich bacterial strain obtained in this invention, Staphylococcus saprophyticus NCSL-YWH2, was deposited with the China General Microbiological Culture Collection Center (CGMCC) on September 9, 2025, with accession number CGMCC No. 35847. The address of the depository is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Detailed Implementation
[0029] The present invention will be further described in detail below through specific embodiments. It should be understood that the following embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.
[0030] Unless otherwise specified, the experimental materials, operating methods, etc. used in the following embodiments can be obtained through conventional commercial channels, are known in the technical field, or are performed in accordance with the product instructions.
[0031] The *Staphylococcus saprophyticus* NCSL-YWH2 obtained in this invention, after being cultured on LB medium, produces colonies that are round, smooth, raised, and opaque white. Figure 1 As shown.
[0032] Figure 2 Phylogenetic tree of Staphylococcus saprophyticus NCSL-YWH2 strain.
[0033] The *Staphylococcus saprophyticus* NCSL-YWH2 strain of the present invention is a bacterium with lithium enrichment capabilities. It can survive and enrich lithium ions in environments with different lithium contents (0-100 g / L lithium chloride), and is applicable to the extraction of lithium ions from water resources with low lithium content, such as geothermal brine and seawater.
[0034] The nucleotide sequence of the 16S rDNA of Staphylococcus saprophyticus NCSL-YWH2 of the present invention preferably includes the sequence shown in SEQ ID NO.1.
[0035] Example 1: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in simulated lithium-containing liquid.
[0036] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (tryptone 10 g / L, yeast extract 5 g / L, sodium chloride 10 g / L) in a clean bench and activated in a 250 mL Erlenmeyer flask at 10 °C for 3 days at a shaking speed of 100 rpm until the logarithmic growth phase was reached.
[0037] Step 2: Inoculate the activated strain into 100 mL of sterile liquid culture medium (1 g / L glucose, 0.2 g / L ammonium chloride, 0.05 g / L potassium dihydrogen phosphate, 0.2 g / L magnesium sulfate heptahydrate, 0 g / L lithium chloride, and deionized water to a final volume of 1 L) at a 10% (v / v) in a clean bench. Then, carry out the culture in a 250 mL Erlenmeyer flask at 10°C for 10 days with a shaker speed of 100 rpm.
[0038] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0039] Step 4: Use the direct application strategy, that is, resuspend the bacterial pellet and add it to 100 mL of the simulated lithium-containing liquid system to be enriched at an inoculation rate of 1 g / L. Enrich in a 250 mL Erlenmeyer flask for 1 h at a culture temperature of 20℃ and a shaker speed of 160 rpm.
[0040] The lithium ion concentration in the simulated lithium-containing liquid was 0.20 mg / L. After 1 h, the lithium ion concentration in the solution was 0.01 mg / L, with a removal rate of 95%, and the enrichment amount in the bacteria was 0.19 mg / g stem cells.
[0041] Example 2: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in simulated lithium-containing liquids
[0042] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (tryptone 10 g / L, yeast extract 5 g / L, sodium chloride 10 g / L) in a clean bench and activated in a 250 mL Erlenmeyer flask at 35°C for 1 day at a shaking speed of 250 rpm until the logarithmic growth phase was reached.
[0043] Step 2: Inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (100 g / L glucose, 20 g / L ammonium chloride, 8 g / L potassium dihydrogen phosphate, 5 g / L magnesium sulfate heptahydrate, 100 g / L lithium chloride, and deionized water to a final volume of 1 L) in a clean bench. Then, carry out the culture in a 250 mL Erlenmeyer flask at 35°C for 1 day with a shaker speed of 250 rpm.
[0044] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0045] Step 4: Use the direct application strategy, that is, resuspend the bacterial precipitate and add it to 100 mL of the simulated lithium-containing liquid system to be enriched at an inoculation rate of 1 g / L. Enrich in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0046] The lithium ion concentration in the simulated lithium-containing liquid was 2000 mg / L. After 24 h, the lithium ion concentration in the solution was 1880 mg / L, with a removal rate of 6%, and the enrichment amount in the bacteria was 120 mg / g stem cells.
[0047] Example 3: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in simulated lithium-containing liquids.
[0048] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0049] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0050] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0051] Step 4: Use the direct application strategy, that is, resuspend the bacterial precipitate and add it to 100 mL of the simulated lithium-containing liquid system to be enriched at an inoculation rate of 1 g / L. Enrich in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0052] The lithium ion concentration in the simulated lithium-containing liquid was 50 mg / L. After 24 h, the lithium ion concentration in the solution was 30 mg / L, with a removal rate of 40%, and the enrichment amount in the bacteria was 20 mg / g stem cells.
[0053] Example 4: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in salt lake brine systems
[0054] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (tryptone 10 g / L, yeast extract 5 g / L, sodium chloride 10 g / L) in a clean bench and activated in a 250 mL Erlenmeyer flask at 10 °C for 3 days at a shaking speed of 100 rpm until the logarithmic growth phase was reached.
[0055] Step 2: Inoculate the activated strain into 100 mL of sterile liquid culture medium (1 g / L glucose, 0.2 g / L ammonium chloride, 0.05 g / L potassium dihydrogen phosphate, 0.2 g / L magnesium sulfate heptahydrate, 200 mg / L lithium chloride, and deionized water to a final volume of 1 L) at a 10% (v / v) in a clean bench. Then, carry out the culture in a 250 mL Erlenmeyer flask at 10°C for 10 days with a shaker speed of 100 rpm.
[0056] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0057] Step 4: The direct use strategy was adopted, that is, the bacterial precipitate was resuspended and added to 100 mL of the brine system to be enriched (Qinghai Chaka Salt Lake) at an inoculation rate of 1 g / L. The mixture was then enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0058] The lithium ion concentration in the salt lake brine was 184 mg / L. After 24 h, the lithium ion concentration in the solution was 150 mg / L, with a removal rate of 18%. The enrichment amount in the bacteria was 32.8 mg / g stem cells.
[0059] Example 5: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in geothermal hot spring water.
[0060] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0061] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterile liquid culture medium (10 g / L glycerol, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0062] Step 3: Using the strategy of attaching to the surface of the inorganic carrier, add sterile activated carbon carrier to the conical flask and continue culturing for 1 day.
[0063] Step 4: Filter and wash the inorganic carrier loaded with bacteria, and add it to 100 mL of the geothermal hot spring water system (Yangbajing geothermal area, Tibet) with a carrier inoculation amount of 1 g / L of bacterial equivalent. Enrich in 250 mL Erlenmeyer flasks for 24 h at a culture temperature of 30℃ and a shaking speed of 160 rpm.
[0064] The lithium ion concentration in the geothermal hot spring water was 2.4 mg / L. After 24 hours, the lithium ion concentration in the solution was 1.03 mg / L, with a removal rate of 57%. The enrichment amount in the bacteria was 1.36 mg / g stem cells.
[0065] Example 6: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in geothermal well water
[0066] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0067] Step 2: Inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L sucrose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L) in a clean bench. Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0068] Step 3: Using the strategy of attaching to the surface of the organic polymer carrier, add sterile polyurethane carrier to the conical flask and continue culturing for 1 day.
[0069] Step 4: Filter and wash the organic polymer carrier loaded with bacteria, add 1 g / L of bacterial inoculum to 100 mL of the geothermal well water system to be enriched (Yangbajing geothermal area, Tibet), enrich for 24 h in a 250 mL Erlenmeyer flask, culture temperature 30℃, shaker speed 160 rpm.
[0070] The lithium ion concentration in the geothermal well water was 9.2 mg / L. After 24 h, the lithium ion concentration in the solution was 5.3 mg / L, with a removal rate of 42%. The enrichment amount in the bacteria was 3.8 mg / g stem cells.
[0071] Example 7: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in produced water of oil and gas fields.
[0072] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (tryptone 10 g / L, yeast extract 5 g / L, sodium chloride 10 g / L) in a clean bench and activated in a 250 mL Erlenmeyer flask at 35°C for 1 day at a shaking speed of 250 rpm until the logarithmic growth phase was reached.
[0073] Step 2: Inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (molasses 100 g / L, ammonium chloride 20 g / L, potassium dihydrogen phosphate 8 g / L, magnesium sulfate heptahydrate 5 g / L, lithium chloride 50 mg / L, and deionized water to a final volume of 1 L) in a clean bench. Then, carry out the culture in a 250 mL Erlenmeyer flask at 35℃ for 1 day with a shaker speed of 250 rpm.
[0074] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0075] Step 4: The direct use strategy was adopted, that is, the bacterial precipitate was resuspended and added to 100 mL of the produced water system of the oil and gas field to be enriched (produced water of the Kuqa foreland gas field in Xinjiang) at an inoculation amount of 1 g / L. The mixture was then enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 35℃ and a shaking speed of 160 rpm.
[0076] The lithium ion concentration in the produced water from the oil and gas field was 38 mg / L. After 24 hours, the lithium ion concentration in the solution was 22.8 mg / L, with a removal rate of 40%. The enrichment amount in the bacteria was 15 mg / g stem cells.
[0077] Example 8: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in lithium-containing mineral water.
[0078] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0079] Step 2: Inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L sucrose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L) in a clean bench. Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0080] Step 3: Using the strategy of adhering to the surface of the composite carrier, add sterile magnetic polyvinyl alcohol formaldehyde (Fe3O4@SiO2 / CSF-PVFM) to the conical flask and continue to incubate for 1 day.
[0081] Step 4: Filter and wash the composite carrier loaded with bacteria, add 1 g / L of bacterial inoculum to 100 mL of the lithium-containing mineral water system (mineral water from Yutian County, Xinjiang), enrich in a 250 mL Erlenmeyer flask for 24 h, culture at 30℃ and shaker speed of 160 rpm.
[0082] The lithium-containing mineral water had a lithium ion concentration of 1.76 mg / L, and after 24 h, the lithium ion concentration in the solution was 0.62 mg / L, with a removal rate of 65%, and the enrichment amount in the bacteria was 1.14 mg / g stem cells.
[0083] Example 9: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in seawater.
[0084] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0085] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0086] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0087] Step 4: Use the direct application strategy, that is, resuspend the bacterial pellet and add it to 100 mL of the seawater system (Qingdao) to be enriched at an inoculation rate of 1 g / L. Enrich in 250 mL Erlenmeyer flasks for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0088] The lithium ion concentration in the seawater was 0.20 mg / L, and after 24 h, the lithium ion concentration in the solution was 0.17 mg / L, with a removal rate of 15%, and the enrichment amount in the bacteria was 0.03 mg / g stem cells.
[0089] Example 10: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in lithium ore leaching solution
[0090] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0091] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L starch, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0092] Step 3: Using a strategy based on natural polymer materials to prepare encapsulated particles, the expanded bacterial cell suspension was centrifuged at 3000 rpm for 5 minutes to obtain a mixed liquid suspended solids concentration (MLSS) of approximately 2 × 10⁻⁶. 4 A concentrated bacterial culture of mg / L was added. A 3% sodium alginate solution was added, and after stirring until homogeneous, the solution was dripped into a 4% calcium chloride solution via a peristaltic pump for cross-linking over 4 hours.
[0093] Step 4: The embedded particles prepared using natural polymer materials were filtered and washed, and then added to 100 mL of the lithium ore leaching solution system (the leaching solution obtained by roasting and leaching lithium mica ore with sodium carbonate in a certain place in Jiangxi) at an inoculation amount of 2 g / L. The solution was then enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaking speed of 160 rpm.
[0094] The lithium ion concentration in the lithium ore leaching solution was 1760 mg / L, and after 24 h, the lithium ion concentration in the solution was 1655 mg / L, with a removal rate of 6%, and the enrichment amount in the bacteria was 52 mg / g stem cells.
[0095] Example 11: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in lithium battery production wastewater
[0096] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0097] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (5 g / L glucose, 5 g / L sucrose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 100 g / L lithium chloride, and bring the volume to 1 L with deionized water). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0098] Step 3: Using a strategy based on synthetic polymer materials to prepare encapsulated particles, the expanded bacterial cell suspension was centrifuged at 3000 rpm for 5 minutes to obtain a mixed liquid suspended solids concentration (MLSS) of approximately 2 × 10⁻⁶. 4 A concentrated bacterial culture of mg / L was added. A polyurethane prepolymer emulsion was then added, followed by 0.5% tetramethylethylenediamine catalyst and 1% potassium persulfate initiator. The reaction was carried out at 25°C for 10 minutes.
[0099] Step 4: The embedded particles prepared using synthetic polymer materials were filtered and washed, and then added to 100 mL of lithium battery production wastewater system (the ternary cathode material production wastewater of a new energy company) at an inoculation rate of 1 g / L. The mixture was then enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0100] The lithium ion concentration in the lithium battery production wastewater was 1700 mg / L. After 24 hours, the lithium ion concentration in the solution was 1569 mg / L, with a removal rate of 7.7%. The enrichment amount in the bacteria was 120 mg / g stem cells.
[0101] Example 12: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in wastewater from the recycling process of decommissioned lithium-ion batteries.
[0102] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0103] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0104] Step 3: Using a strategy based on composite materials to prepare embedded particles, the expanded bacterial cell suspension was centrifuged at 3000 rpm for 5 minutes to obtain a mixed liquid suspended solids concentration (MLSS) of approximately 2 × 10⁻⁶. 4 A concentrated bacterial solution of mg / L was prepared. 10% polyvinyl alcohol was completely dissolved at 95℃, and 1% sodium alginate was added and stirred thoroughly until completely dissolved. The solution was then allowed to stand in a water bath at 35℃ to remove foam. 2% zeolite was added and mixed evenly. The concentrated bacterial solution was then mixed evenly with the polyvinyl alcohol-sodium alginate-zeolite solution. The solution was then added dropwise to a saturated boric acid solution of CaCl2 and crosslinked at 4℃ for 24 h.
[0105] Step 4: The embedded particles prepared using composite materials were filtered and washed, and then added to 100 mL of wastewater system from the recycling process of retired lithium-ion batteries (the leachate obtained after grinding and acid leaching of retired lithium iron phosphate batteries from a battery recycling company) at an inoculation rate of 1 g / L. The mixture was then enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaking speed of 160 rpm.
[0106] The lithium ion concentration in the wastewater from the recycling process of the decommissioned lithium-ion batteries was 1400 mg / L. After 24 hours, the lithium ion concentration in the solution was 1288 mg / L, with a removal rate of 8%. The enrichment amount in the bacteria was 108 mg / g stem cells.
[0107] Example 13: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in wastewater recovery in the glass industry.
[0108] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0109] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0110] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0111] Step 4: Use the direct application strategy, that is, resuspend the bacterial precipitate and add it to 100 mL of the glass industry wastewater system (the leachate after calcination and alkali leaching of lithium aluminum silicate glass) at an inoculation rate of 1 g / L. Then enrich it in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaking speed of 160 rpm.
[0112] The lithium ion concentration in the recycled wastewater from the glass industry was 1368 mg / L. After 24 hours, the lithium ion concentration in the solution was 1257 mg / L, with a removal rate of 8.1%. The enrichment amount in the bacteria was 102 mg / g stem cells.
[0113] Example 14: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in wastewater recovery in the ceramics industry.
[0114] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0115] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0116] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0117] Step 4: Use the direct application strategy, that is, resuspend the bacterial precipitate and add it to 100 mL of the ceramic industry wastewater system to be enriched (the leachate after calcination of lithium aluminate ceramics) at an inoculation rate of 1 g / L. Enrich in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaking speed of 160 rpm.
[0118] The lithium ion concentration in the recycled wastewater from the ceramic industry was 880 mg / L. After 24 hours, the lithium ion concentration in the solution was 783 mg / L, with a removal rate of 11%. The enrichment amount in the bacteria was 93 mg / g stem cells.
[0119] Example 15: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in wastewater from lithium-based lubricant industrial production.
[0120] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0121] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0122] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0123] Step 4: The direct use strategy was adopted, that is, the bacterial precipitate was resuspended and added to 100 mL of the lithium-based lubricant industrial wastewater system to be enriched at an inoculation rate of 1 g / L. The mixture was then enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0124] The lithium ion concentration in the industrial wastewater from the lithium-based lubricant production was 40 mg / L. After 24 hours, the lithium ion concentration in the solution was 31 mg / L, with a removal rate of 22.5%. The enrichment amount in the bacteria was 9 mg / g stem cells.
[0125] Example 16: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in wastewater from lithium alloy industrial production.
[0126] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0127] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0128] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0129] Step 4: Use the direct application strategy, that is, resuspend the bacterial precipitate and add it to 100 mL of the lithium alloy industrial wastewater system (aluminum-lithium alloy processing dust leachate) at an inoculation rate of 1 g / L. Enrich in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0130] The lithium ion concentration in the industrial wastewater containing lithium alloys was 21 mg / L. After 24 hours, the lithium ion concentration in the solution was 16 mg / L, with a removal rate of 24%. The enrichment amount in the bacteria was 4.6 mg / g stem cells.
[0131] Example 17: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in wastewater after residual liquid adsorption treatment in the industrial production of medical lithium carbonate.
[0132] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0133] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0134] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0135] Step 4: The direct use strategy was adopted, that is, the bacterial precipitate was resuspended and added at an inoculum of 1 g / L to 100 mL of the wastewater system after adsorption treatment in the industrial production process of medical lithium carbonate (the tail water after adsorption by manganese ion sieve after lithium carbonate precipitation). The mixture was enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaking speed of 160 rpm.
[0136] The lithium ion concentration in the wastewater after adsorption treatment in the industrial production process of medical lithium carbonate was 8 mg / L. After 24 h, the lithium ion concentration in the solution was 4.3 mg / L, with a removal rate of 46.3%. The enrichment amount in the bacteria was 3.7 mg / g stem cells.
[0137] Example 18: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in produced water from simulated lithium-bearing oil and gas fields.
[0138] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0139] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0140] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0141] Step 4: Use the direct application strategy, that is, resuspend the bacterial precipitate and add it to 100 mL of the simulated lithium-containing oil and gas field produced water system at an inoculation rate of 5 g / L. Then enrich it in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaking speed of 160 rpm.
[0142] The lithium ion concentration in the produced water from the simulated lithium-containing oil and gas field was 42 mg / L. After 24 h, the lithium ion concentration in the solution was 2.1 mg / L, with a removal rate of 95%, and the enrichment amount in the bacteria was 7.98 mg / g stem cells.
[0143] Example 19: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in wastewater after solvent extraction treatment in lithium battery production.
[0144] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0145] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0146] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0147] Step 4: The direct use strategy was adopted, that is, the bacterial precipitate was resuspended and added at an inoculum of 1 g / L to 100 mL of the tail water system after solvent extraction treatment in the lithium battery production process (the remaining liquid after TBP-FeCl3 extraction of the ternary cathode material production wastewater of a new energy company). The mixture was then enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0148] The lithium ion concentration in the tailwater after solvent extraction treatment in the lithium battery production process was 22.4 mg / L. After 24 h, the lithium ion concentration in the solution was 15.8 mg / L, with a removal rate of 29.5%. The enrichment amount in the bacteria was 6.5 mg / g stem cells.
[0149] Example 20: Cultivation of Staphylococcus saprophyticus NCSL-YWH2 and its application in the treatment of tailwater after adsorption-membrane coupling in lithium battery recycling.
[0150] Step 1: The strain was inoculated into 100 mL of sterilized LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) in a clean bench and activated in a 250 mL Erlenmeyer flask at 30 °C for 1 day at a shaking speed of 180 rpm until the logarithmic growth phase was reached.
[0151] Step 2: In a clean bench, inoculate the activated strain at a rate of 10% (v / v) into 100 mL of sterilized liquid culture medium (10 g / L glucose, 1 g / L ammonium chloride, 1 g / L potassium dihydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 50 mg / L lithium chloride, and deionized water to a final volume of 1 L). Then, carry out the culture in a 250 mL Erlenmeyer flask at 30°C for 1 day with a shaker speed of 180 rpm.
[0152] Step 3: Centrifuge the expanded bacterial suspension at 3000 rpm for 10 minutes and collect the bacterial pellet.
[0153] Step 4: The direct application strategy was adopted, that is, the bacterial precipitate was resuspended and added at an inoculum of 1 g / L to 100 mL of the tail water system after adsorption-membrane coupling treatment in the lithium battery recycling process (the leachate obtained by grinding and acid leaching of retired lithium iron phosphate batteries from a battery recycling company, which was then treated by aluminum adsorbent-nanofiltration membrane coupling). The mixture was enriched in a 250 mL Erlenmeyer flask for 24 h at a culture temperature of 30℃ and a shaker speed of 160 rpm.
[0154] The lithium ion concentration in the tailwater after adsorption-membrane coupling treatment during the lithium battery recycling process was 0.31 mg / L, and the lithium ion concentration in the solution after 24 h was 0.25 mg / L, with a removal rate of 19.4% and an enrichment amount of 0.06 mg / g stem cells in the bacteria.
[0155] Based on the results of Examples 1-20 above, it can be seen that the Staphylococcus saprophyticus NCSL-YWH2 of the present invention can be used to treat various types of lithium-containing water bodies, including salt lake brine, geothermal hot springs and hot brines, produced water from oil and gas fields, lithium-containing mineral water, seawater, lithium ore leachate, lithium battery production wastewater, wastewater from the recycling process of retired lithium-ion batteries, wastewater from the industrial production and recycling processes of lithium-containing chemicals and materials such as glass ceramics, lubricants, alloys, and pharmaceuticals, simulate lithium-containing liquids, and one or more of the liquids generated during the processing of the above-mentioned lithium-containing liquids. It can effectively enrich lithium ions in water bodies, thereby realizing the enrichment, extraction, and recovery of lithium resources.
[0156] The above description is merely a preferred embodiment for explaining the present invention and is not intended to limit the present invention in any way. Therefore, any modifications or changes made to the present invention under the same inventive spirit should still be included within the scope of protection intended by the present invention.
Claims
1. A strain of saprophytic Staphylococcus aureus ( Staphylococcus saprophyticus NCSL-YWH2, characterized in that, Its accession number is CGMCC No.35847.
2. The *Staphylococcus saprophyticus* according to claim 1 ( Staphylococcus saprophyticus NCSL-YWH2, characterized in that, Its 16S rDNA nucleotide sequence is shown in SEQ ID NO.
1.
3. The *Staphylococcus saprophyticus* as described in claim 1 or 2 ( Staphylococcus saprophyticus The method for cultivating NCSL-YWH2 is characterized by, include: Activation culture: The strain was inoculated into LB liquid medium for activation culture at a temperature of 10-35℃ for 1-3 days and a shaking speed of 100-250 rpm until the logarithmic growth phase was reached. Expanded culture: The activated strain was inoculated into liquid culture medium at an inoculation rate of 10% (v / v) for expanded culture. The culture temperature was 10-35℃, the culture time was 1-10 days, and the shaking speed was 100-250 rpm.
4. The cultivation method according to claim 3, characterized in that, The LB liquid culture medium used in the activation culture consists of: 10 g / L tryptone, 5 g / L yeast extract, and 10 g / L sodium chloride.
5. The cultivation method according to claim 3, characterized in that, The liquid culture medium used in the extended culture consists of: carbon source 1–100 g / L, ammonium chloride 0.2–20 g / L, potassium dihydrogen phosphate 0.05–8 g / L, magnesium sulfate heptahydrate 0.2–5 g / L, and lithium chloride 0–100 g / L. The carbon source is selected from one or more combinations of glucose, starch, sucrose, glycerol, and molasses.
6. The *Staphylococcus saprophyticus* as described in claim 1 or 2 ( Staphylococcus saprophyticus The application of NCSL-YWH2 is characterized by, Used to enrich lithium in lithium-containing liquids.
7. The application according to claim 6, characterized in that, The lithium element enriched in the lithium-containing liquid is obtained by removing Staphylococcus saprophyticus (Staphylococcus aureus) Staphylococcus saprophyticus NCSL-YWH2 cells can be used directly, attached to the surface of other functional carriers, or prepared into encapsulated particles.
8. The application according to claim 7, characterized in that, The functional carriers include inorganic carriers, organic polymer carriers, and composite carriers; The embedded particulate materials include natural polymer materials, synthetic polymer materials, and composite materials.
9. The application according to claim 6, characterized in that, The lithium-ion concentration of the lithium-containing liquid ranges from 0.2 to 2000 mg / L.
10. The application according to claim 6, characterized in that, The lithium-containing liquid is selected from one or more of the following: salt lake brine, geothermal hot springs and hot brine, produced water from oil and gas fields, lithium-containing mineral water, seawater, lithium ore leachate, lithium battery production wastewater, wastewater from the recycling process of retired lithium-ion batteries, wastewater from the industrial production and recycling processes of lithium-containing chemicals and materials such as glass ceramics, lubricants, alloys, and pharmaceuticals, simulated lithium-containing liquid, and liquids generated during the processing of the above-mentioned lithium-containing liquids.