WO2020108214A1 - 一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法 - Google Patents

一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法 Download PDF

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WO2020108214A1
WO2020108214A1 PCT/CN2019/114348 CN2019114348W WO2020108214A1 WO 2020108214 A1 WO2020108214 A1 WO 2020108214A1 CN 2019114348 W CN2019114348 W CN 2019114348W WO 2020108214 A1 WO2020108214 A1 WO 2020108214A1
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arsenic
soil
under aerobic
active iron
manganese oxide
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王华伟
王亚楠
孙英杰
高莹
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Qingdao University of Technology
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/08—Reclamation of contaminated soil chemically
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes

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  • the invention belongs to the technical field of environmental protection soil remediation; relates to a method for bacteria-induced active iron-manganese oxide to repair arsenic-contaminated soil under aerobic/microaerobic conditions.
  • Arsenic is a highly toxic metal that can cause skin cancer, bladder cancer, and kidney disease, posing a serious threat to human health.
  • the heavy use of sewage irrigation, industrial sludge and arsenic-containing pesticides in agricultural production has caused serious arsenic pollution in the soil.
  • Plant remediation technology is the most environmentally friendly soil remediation technology.
  • the remediation process is closer to the natural ecology. Being able to purify and beautify the environment is the biggest advantage of plant remediation technology, and it can also increase soil organic matter and fertility.
  • the phytoremediation technology is applicable to soil with large polluted area and light pollution.
  • the investment in plant remediation technology is low, the remediation week is relatively short, and there is no secondary pollution. Studies have shown that most arsenic-rich plants prefer shade and humidity, and these safe and practical dominant plants have not yet been promoted in northern regions.
  • the microbial remediation of arsenic-contaminated soil is to change the bioavailability of arsenic through microbial redox, adsorption, methylation, precipitation and other biochemical reactions of arsenic to achieve the purpose of remediation of arsenic-contaminated soil.
  • the microbial remediation technology of arsenic contaminated soil mainly includes two types: one is biosorption technology, and the other is biooxidation reduction technology. Bioremediation technology is through the fixation of the cell walls of microorganisms.
  • the cell walls of many types of microorganisms such as bacteria, algae and fungi have the ability to combine with metal ions; even some microorganisms will absorb arsenic and use arsenic as a nutrient element necessary for growth and development.
  • Biological redox technology uses microorganisms to change the valence state and activity of arsenic in arsenic-contaminated soil to form intolerant compounds, so as to reduce the toxicity of arsenic in soil and reduce soil pollution.
  • the main problem of the above microbial remediation technology is that the ability to repair arsenic is very limited, and arsenic is prone to migration.
  • the purpose of the present invention is to provide a method for bacteria-induced active iron-manganese oxide to repair arsenic-contaminated soil under aerobic/microaerobic conditions; proceed as follows:
  • Pseudomonas putida P. putida strain MnB1 was inoculated into Pseudomonas putida enrichment medium A at a switching volume of 5-10% by volume, and then shaken at a level of 15-35 degrees Celsius (100-180rpm) , Aerobic enrichment culture for 1-5 days to obtain Pseudomonas putida bacteria liquid;
  • the medium A component is yeast extract powder 0.3-0.8g, hydrolyzed casein 0.2-0.8g, glucose 0.3-0.8g, chlorine Calcium chloride 0.1-0.4g, magnesium sulfate 0.1-0.6g, trace elements 1-5mL, deionized water 1 liter;
  • the selected bacterial strains come from the American Type Bacteria Collection Center, deposit number ATCC23483;
  • step 1) Add the Pseudomonas putida bacteria solution enriched in step 1) to the Pseudomonas putida medium B with a pH of 6.5-7.5 at a volume ratio of 5-10%, and add 0.2-1.8g at the same time /L manganese carbonate, and then cultured at 15-35 degrees Celsius under aerobic conditions for 3-10 days, brown brown flocs appear in the medium, that is, bacterial metabolites are prepared;
  • the components of the medium B are 0.05-0.45g of ferrous ammonium sulfate, 0.1-0.5g of sodium citrate, 0.01-0.3g of yeast extract, 0.01-0.05g of sodium pyrophosphate, and 1 liter of deionized water;
  • step (2) Add 20-80mL of the bioactive iron-manganese oxide prepared in step (2) to 50 grams of arsenic-contaminated soil, stir for 5-30 minutes with a stirrer, and then place it at room temperature (20-40 degrees Celsius) to keep the soil at Incubate for 3-21 days under aerobic/micro-aerobic conditions to allow these active iron-manganese oxides to chemically react with trivalent arsenic or pentavalent arsenic in the soil to convert the exchangeable arsenic into a residue state to be fixed in the soil To achieve the repair effect.
  • the Pseudomonas putida P. putida strain MnB1 purchased through the American Model Strain Collection Center is mixed with Pseudomonas putida medium B containing manganese carbonate at an inoculation amount of 5-10%, and the pH is adjusted to 6.5-7.5 After cultivating at 15-35 degrees Celsius for 3-10 days, the active iron-manganese oxide is obtained.
  • the product is a good soil remediation agent. Adding these biologically active iron and manganese oxides to arsenic-contaminated soil and cultivating them under aerobic/microaerobic conditions for 3-21 days can convert the exchangeable arsenic in the soil to the residue state, so as to achieve the remediation effect.
  • This method selects Pseudomonas putida strains MnB1 capable of dissociating and oxidizing bivalent iron and manganese, enriches the bacteria in an enriched culture medium, and then inoculates the strain into a culture containing ferric iron and manganese Metabolites are cultivated in the base, and the metabolites are mainly bioactive oxides rich in iron and manganese. These bioactive iron and manganese oxides are then added to the arsenic-contaminated soil and cultivated for several days under aerobic/microaerobic conditions Terminate the experiment.
  • These biologically active iron-manganese oxides can effectively convert the exchangeable arsenic to the residual state, thereby effectively fixing the arsenic in the soil and repairing the soil arsenic pollution.
  • the method has simple process, convenient operation, low processing cost, large processing range and no secondary pollution.
  • Figure 1 Scanning electron microscope diagram of bacteria-induced active iron-manganese oxide in Example 1
  • Figure 2 Schematic diagram of the content of water-soluble arsenic before and after repairing with active iron and manganese oxide in Example 1
  • Figure 3 Schematic diagram of the morphological changes of arsenic before and after repairing arsenic with active iron-manganese oxide in Example 1
  • FIG. 4 Example 2 Schematic diagram of changes in water-soluble arsenic content before and after active iron-manganese oxide repair
  • Figure 5 Schematic diagram of the morphological changes of arsenic before and after repairing arsenic with active iron-manganese oxide in Example 2
  • Figure 6 Schematic diagram of the change of water-soluble arsenic content before and after repair of active iron-manganese oxide in Example 3
  • Figure 7 Schematic diagram of the morphological changes of arsenic before and after repairing arsenic with active iron-manganese oxide in Example 3.
  • Pseudomonas putida P. putida strain MnB1 was inoculated into Pseudomonas putida enrichment medium A at a switching volume of 5% by volume, and then aerobic enrichment was performed at 25 degrees Celsius (150rpm) 2 days of cultivation;
  • the components of the medium A are 0.5g of yeast extract powder, 0.5g of hydrolyzed casein, 0.5g of glucose, 0.29g of calcium chloride, 0.5g of magnesium sulfate, 1mL of trace elements, and 1 liter of deionized water;
  • the selected bacterial strains are from the American Type Bacteria Collection Center ATCC, deposit number ATCC23483;
  • the enriched Pseudomonas putida bacterial solution obtained in step 1) was added to the Pseudomonas putida medium B containing manganese carbonate with an initial concentration of 1 g/L and a pH of 6.8 at an inoculation amount of 6% by volume, and then at 25 degrees Celsius , Cultured under aerobic conditions for 4 days, the medium becomes brown and black to produce bacterial metabolites;
  • the components of the medium B are 0.15g of ferrous ammonium sulfate, 0.15g of sodium citrate, 0.075g of yeast extract powder, 0.05g of sodium pyrophosphate, and 1 liter of deionized water.
  • step (2) Add 50mL of the active iron-manganese oxide prepared in step (2) to 50 grams of arsenic-contaminated soil, stir for 30 minutes with a stirrer, and then place it at room temperature (25 degrees Celsius) for 15 days to oxidize these active iron-manganese
  • the chemical reaction between the substance and arsenic will convert the exchangeable arsenic into the residue state and fix it in the soil to achieve the remediation effect.
  • Pseudomonas putida P. putida strain MnB1 was inoculated into Pseudomonas putida enrichment medium A at a transfer volume of 10% by volume, and then aerobic enrichment culture was performed at 25°C (150 rpm) with horizontal shaking 5 days;
  • the medium A consists of 0.5g yeast extract powder, 0.5g hydrolyzed casein, 0.5g glucose, 0.2g calcium chloride, 0.3g magnesium sulfate, 4mL trace elements, and 1 liter deionized water;
  • the selected bacterial strains come from the American Type Bacteria Collection Center, deposit number ATCC23483;
  • step 1) Add the Pseudomonas putida bacterial solution enriched in step 1) to the Pseudomonas putida medium B containing manganese carbonate with an initial concentration of 1.0 g/L and a pH of 6.8 at an inoculation volume of 10% by volume, and then At 30 degrees Celsius, cultured for 7 days under aerobic conditions, brown and black flocs appeared in the medium, that is, bacterial metabolites were prepared;
  • the components of the medium B are 0.35 g of ferrous ammonium sulfate, 0.25 g of sodium citrate, 0.07 g of yeast extract powder, 0.025 g of sodium pyrophosphate, and 1 liter of deionized water;
  • Pseudomonas putida P. putida strain MnB1 was inoculated into Pseudomonas putida enrichment medium A at a transfer volume of 8% by volume, and then aerobic enrichment culture was carried out at 20°C (120 rpm) with horizontal shaking 5 days;
  • the components of medium B are 0.35g of yeast extract powder, 0.35g of hydrolyzed casein, 0.35g of glucose, 0.2g of calcium chloride, 0.5g of magnesium sulfate, 2mL of trace elements, and 1 liter of deionized water;
  • the selected bacterial strains come from the American Type Bacteria Collection Center, deposit number ATCC23483;
  • the enriched Pseudomonas putida bacterial solution obtained in step 1) was added to Pseudomonas putida medium B containing manganese carbonate with an initial concentration of 1.0 g/L and a pH of 7.0 at a volume ratio of 7%.
  • the ingredients are 0.25g ferrous ammonium sulfate, 0.25g sodium citrate, 0.23g yeast extract, 0.02g sodium pyrophosphate, 1 liter deionized water, and then cultured at 30 degrees Celsius under aerobic conditions for 4 days in medium The appearance of brown-black flocs, that is, bacterial metabolites;
  • step (2) Add 40mL of the bioactive iron-manganese oxide prepared in step (2) to 50 grams of arsenic-contaminated soil, stir for 10 minutes with a stirrer, and then place it at room temperature (30 degrees Celsius) to keep the soil aerobic/microaerobic Incubate for 7 days under conditions.
  • the method of the present invention can achieve the effect of remediation of arsenic-contaminated soil.
  • P. putida strain MnB1 converts the ferric manganese in the medium into a mixed metabolite of bacteria-iron-manganese oxide, which is a good soil remediation agent.
  • these metabolites are added to arsenic pollution, it can effectively reduce the mobility and biological effectiveness of arsenic in the soil, thereby fixing the arsenic in the soil.
  • the method has simple process and convenient operation. The treatment cost is low, there is no secondary pollution, and the arsenic-contaminated soil can be repaired.

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Abstract

一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法，该方法选取恶臭假单胞菌（ Pseudomonas putida strain MnB1，ATCC 23483），将该菌种加入到含有碳酸锰的培养基中培养3-10天，直到细菌诱导产生棕黑色絮状产物，而后将代谢产物加入到砷污染土壤中，在好氧/微好氧条件下培养3-21天。在细菌代谢产物的作用下，可以将土壤中可交换态的砷转化为残渣态的砷，降低砷在土壤中的迁移和生物有效性，有效固定土壤中的砷。该方法操作简单，处理成本低，适用范围广，无二次污染。

Description

一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法 技术领域
本发明属于环境保护土壤修复处理技术领域；涉及一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法。
技术背景
砷(As)是一种高毒类金属，会引起皮肤癌、膀胱癌以及肾脏疾病，对人体健康具有严重威胁。由于污水灌溉、工业污泥及含砷农药等在农业生产中的大量使用，造成土壤严重的砷污染。研究表明，砷污染土壤中主要以无机的三价砷和五价砷为主，而在土壤环境中，三价砷的毒性的迁移性均高于五价砷。
砷污染土壤修复技术主要有物理-化学修复法、植物修复法和微生物修复法三大类。传统的物理-化学修复技术通常投加量大、成本高、易产生二次污染，不适用大面积砷污染土壤的修复。
植物修复技术是最环保的土壤修复技术，修复过程更接近自然生态，能够净化与美化环境是植物修复技术的最大优势，而且还能增加土壤有机质和肥力。植物修复技术适用于污染面积大，轻度污染的土壤。而且植物修复技术的投资少、修复周比较短期也短并且没有二次污染。研究表明，大多数砷富集植物喜阴喜湿，这些安全并实用的优势植物尚无法在北方地区得到推广。
砷污染土壤的微生物修复作用是通过微生物对砷的氧化还原、吸附、甲基化、沉淀等生化反应来改变砷的生物有效性，来实现修复砷污染土壤的目的。砷污染土壤的微生物修复技术主要包括两种：一个是生物吸附技术，另一个是生物氧化还原技术。生物修复技术是通过微生物细胞壁的固定作用，细菌、藻类、真菌等多类微生物的细胞壁都具有与金属离子结合的能力；甚至有的微生物会吸收砷，把砷作为生长发育所必须营养元素。生物氧化还原技术是利用微生物可以改变砷污染土壤中砷的离子价态和活性，使其形成不容的化合物，从而达到降低土壤中砷的毒性，减轻土壤污染的目的。上述微生物修复技术存在的主要问题是对砷修复能力十分有限，且砷极易发生迁移。
发明内容
本发明的目的在于，提供一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法；按下列步骤进行：
1)细菌菌株的富集培养：
将恶臭假单胞菌P.putida strain MnB1按体积比5-10％的转接量接种于恶臭假单胞菌富集 培养基A中，然后于15-35摄氏度水平震荡下(100-180rpm)，好氧富集培养1-5天，得到恶臭假单细胞菌菌液；所述培养基A成分为酵母浸粉0.3-0.8g，水解酪蛋白0.2-0.8g，葡萄糖0.3-0.8g，氯化钙0.1-0.4g，硫酸镁0.1-0.6g，微量元素1-5mL，去离子水1升；
选用的细菌菌株来自于美国模式菌收集中心，保藏号ATCC 23483；
2)活性铁锰氧化物的制备：
将步骤1)富集得到的恶臭假单胞菌菌液按体积比5-10％的接种量加入到pH为6.5-7.5的恶臭假单胞菌培养基B中，同时投加0.2-1.8g/L的碳酸锰，然后于15-35摄氏度，好氧条件下培养3-10天，培养基中出现棕黑色絮状体，即制得细菌代谢产物；
所述培养基B的成分为硫酸亚铁铵0.05-0.45g，柠檬酸钠0.1-0.5g，酵母浸粉0.01-0.3g，焦磷酸钠0.01-0.05g，去离子水1升；
3)修复过程
将20-80mL步骤(2)制得的生物活性铁锰氧化物加入到50克砷污染土壤中，搅拌器搅拌5-30分钟，然后置于室温条件下(20-40摄氏度)，保持土壤为好氧/微好氧条件下培养3-21天，让这些活性铁锰氧化物与土壤中的三价砷或五价砷发生化学反应，将可交换的砷转化为残渣态从而固定在土壤中，达到修复效果。
有益效果：
本发明通过美国模式菌种收集中心购买的恶臭假单胞菌P.putida strain MnB1，按5-10％的接种量与含有碳酸锰的恶臭假单胞菌培养基B混合，调剂pH 6.5-7.5，在15-35摄氏度下培养3-10天即获得活性铁锰氧化物，该产物是一种良好的土壤修复剂。将这些生物活性铁锰氧化物加入到砷污染土壤中，好氧/微好氧条件下培养3-21天，可以将土壤中可交换态的砷转化为残渣态，从而达到修复效果。
该方法选取能够将二价铁、锰异化氧化的恶臭假单胞菌Pseudomonas putida strain MnB1，将该菌在富集培养基中富集培养，然后将该菌株接种到含有二价铁、锰的培养基中培养得到代谢产物，代谢产物主要为富含铁、锰的生物活性氧化物，而后将这些生物活性铁锰氧化物加入到砷污染土壤中，在好氧/微好氧条件下培养数天后终止实验。这些生物活性铁锰氧化物能够有效将可交换态的砷转化为残渣态，进而有效固定土壤中的砷，修复土壤砷污染。该方法工艺简单、操作方便、处理成本低、处理范围大、无二次污染。
附图说明
图1：实施例1细菌诱导活性铁锰氧化物的扫描电子显微镜示意图
图2：实施例1活性铁锰氧化物修复前后水溶性砷含量变化示意图
图3：实施例1活性铁锰氧化物修复砷前后砷的存在形态变化示意图
图4：实施例2活性铁锰氧化物修复前后水溶性砷含量变化示意图
图5：实施例2活性铁锰氧化物修复砷前后砷的存在形态变化示意图
图6：实施例3活性铁锰氧化物修复前后水溶性砷含量变化示意图
图7：实施例3活性铁锰氧化物修复砷前后砷的存在形态变化示意图
具体实施方式
以下通过实验案例对本发明作进一步说明，而非限制本发明。
实施案例1
1)细菌的富集培养
将该恶臭假单胞菌P.putida strain MnB1按体积比5％的转接量接种于恶臭假单胞菌富集培养基A中，然后于25摄氏度水平震荡下(150rpm)，好氧富集培养2天；
所述培养基A成分为酵母浸粉0.5g，水解酪蛋白0.5g，葡萄糖0.5g，氯化钙0.29g，硫酸镁0.5g，微量元素1mL，去离子水1升；
选用的细菌菌株来自于美国模式菌收集中心ATCC，保藏号ATCC 23483；
2)活性代谢产物的制备：
将步骤1)富集得到的恶臭假单胞菌菌液按体积比6％的接种量加入含碳酸锰初始浓度为1g/L pH为6.8的恶臭假单胞菌培养基B，然后于25摄氏度，好氧条件下培养4天，培养基变成棕黑色即制得细菌代谢产物；
所述培养基B的成分为硫酸亚铁铵0.15g，柠檬酸钠0.15g，酵母浸粉0.075g，焦磷酸钠0.05g，去离子水1升，
3)修复过程
将50mL步骤(2)制得的活性铁锰氧化物加入到50克砷污染土壤中，搅拌器搅拌30分钟，然后置于室温条件下(25摄氏度)中放置15天，让这些活性铁锰氧化物与砷发生化学反应，将可交换的砷转化为残渣态从而固定在土壤中，达到修复效果。
我们对细菌诱导产生的活性铁锰氧化物进行微观结构观察(附图1)，可知，这些活性铁锰氧化物主要为未反应的细菌菌株和碳酸锰以及无定型或弱晶型的铁锰氧化物混合物为主。由附图2可知，当这些铁锰氧化物投加到砷土壤中时，水溶性砷含量明显降低，由未修复时的2.197mg/kg降至0.715mg/kg，修复效率为67.51％。此外，我们对土壤中砷的存在形态进行分析(附图3)，结果表明，未修复时可交换态砷为4.4％，而修复后可交换态砷的百分比降至0.94％，残渣态则由未修复的69.69％增加至77.88％。
实施案例2
1)细菌菌株的富集培养：
将恶臭假单胞菌P.putida strain MnB1按体积比10％的转接量接种于恶臭假单胞菌富集培养基A中，然后于25摄氏度水平震荡下(150rpm)，好氧富集培养5天；
培养基A成分为酵母浸粉0.5g，水解酪蛋白0.5g，葡萄糖0.5g，氯化钙0.2g，硫酸镁0.3g，微量元素4mL，去离子水1升；
选用的细菌菌株来自于美国模式菌收集中心，保藏号ATCC 23483；
2)活性铁锰氧化物的制备：
将步骤1)富集得到的恶臭假单胞菌菌液按体积比10％的接种量加入含碳酸锰初始浓度为1.0g/L，pH为6.8的恶臭假单胞菌培养基B，然后于30摄氏度，好氧条件下培养7天，培养基中出现棕黑色絮状体，即制得细菌代谢产物；
培养基B的成分为硫酸亚铁铵0.35g，柠檬酸钠0.25g，酵母浸粉0.07g，焦磷酸钠0.025g，去离子水1升；
3)修复过程
将20mL步骤(2)制得的生物活性铁锰氧化物加入到50克砷污染土壤中，搅拌器搅拌10分钟，然后置于室温条件下(30摄氏度)，保持土壤为好氧/微好氧条件下培养15天。水溶性砷含量明显降低，由未修复时的2.197mg/kg降至0.715mg/kg，修复效率为67.51％(附图4)。土壤中砷的存在形态分析结果表明，未修复时可交换态砷为4.4％，而修复后可交换态砷的百分比降至1.14％，残渣态则由未修复的69.69％增加至75.88％(附图5)。
实施案例3
1)细菌菌株的富集培养：
将恶臭假单胞菌P.putida strain MnB1按体积比8％的转接量接种于恶臭假单胞菌富集培养基A中，然后于20摄氏度水平震荡下(120rpm)，好氧富集培养5天；
培养基B成分为酵母浸粉0.35g，水解酪蛋白0.35g，葡萄糖0.35g，氯化钙0.2g，硫酸镁0.5g，微量元素2mL，去离子水1升；
选用的细菌菌株来自于美国模式菌收集中心，保藏号ATCC 23483；
2)活性铁锰氧化物的制备：
将步骤1)富集得到的恶臭假单胞菌菌液按体积比7％的接种量加入含碳酸锰初始浓度为1.0g/L，pH为7.0的恶臭假单胞菌培养基B，培养基的成分为硫酸亚铁铵0.25g，柠檬酸钠0.25g，酵母浸粉0.23g，焦磷酸钠0.02g，去离子水1升，然后于30摄氏度，好氧条件下培养4天，培养基中出现棕黑色絮状体，即制得细菌代谢产物；
3)修复过程
将40mL步骤(2)制得的生物活性铁锰氧化物加入到50克砷污染土壤中，搅拌器搅拌10分钟，然后置于室温条件下(30摄氏度)，保持土壤为好氧/微好氧条件下培养7天。
水溶性砷含量明显降低，由未修复时的2.197mg/kg降至0.60mg/kg，修复效率为72.69％(附图6)。砷的存在形态分析结果表明，未修复时可交换态砷为4.4％，而修复后可交换态砷的百分比降至1.00％，残渣态则由未修复的69.69％增加至76.52％(附图7)。
从上述实施例可以看出：通过本发明所述方法可达到修复砷污染土壤的效果。在好氧条件下，恶臭假单胞菌P.putida strain MnB1将培养基中二价铁锰转化为细菌-铁锰氧化物的混合代谢产物，这些代谢产物是一种良好的土壤修复剂。将这些代谢产物添加到砷污染中时，可以有效降低土壤中砷的可迁移性和生物有效性，从而固定土壤中的砷。该方法工艺简单，操作方便。处理成本低，无二次污染，能够实现砷污染土壤的修复。

Claims (3)

  1. 一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法，其特征在于；通过下列步骤进行：
    1)细菌菌株的富集培养：
    将恶臭假单胞菌Pseudomonas putida strain MnB1按体积比5-10％的转接量接种于恶臭假单胞菌富集培养基A中，然后于15-35摄氏度水平震荡下(100-180rpm)，好氧富集培养1-5天；
    2)活性铁锰氧化物的制备：
    将步骤1)富集得到的恶臭假单胞菌菌液按体积比5-10％的接种量pH为6.5-7.5的恶臭假单胞菌培养基B中，同时投加0.2-1.8g/L的碳酸锰，然后于15-35摄氏度，好氧条件下培养3-10天，培养基中产生棕黑色絮状体，即制得活性铁锰氧化物；
    3)修复过程
    将20-80mL步骤(2)制得的生物活性铁锰氧化物加入到50克砷污染土壤中，搅拌器搅拌5-30分钟，然后置于室温条件下(20-40摄氏度)，保持土壤为好氧/微好氧条件下放置3-12天，让这些活性铁锰氧化物与土壤中的三价砷或五价砷发生化学反应。
  2. 如权利要求1所述的好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法，其特征在于；
    所述培养基A成分为酵母浸粉0.3-0.8g，水解酪蛋白0.2-0.8g，葡萄糖0.3-0.8g，氯化钙0.1-0.4g，硫酸镁0.1-0.6g，微量元素1-5mL，去离子水1升，
    所述培养基B的成分为硫酸亚铁铵0.05-0.45g，柠檬酸钠0.1-0.5g，酵母浸粉0.01-0.3g，焦磷酸钠0.01-0.05g，去离子水1升。
  3. 如权利要求1所述的好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法，其特征在于；步骤1)中选用的细菌菌株来自于美国模式菌收集中心(ATCC)，保藏号ATCC 23483。
PCT/CN2019/114348 2018-11-28 2019-10-30 一种好氧/微好氧条件下细菌诱导活性铁锰氧化物修复砷污染土壤的方法 Ceased WO2020108214A1 (zh)

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