CN106587506B - 一种去除含砷污水污染物的方法 - Google Patents
一种去除含砷污水污染物的方法 Download PDFInfo
- Publication number
- CN106587506B CN106587506B CN201611187827.3A CN201611187827A CN106587506B CN 106587506 B CN106587506 B CN 106587506B CN 201611187827 A CN201611187827 A CN 201611187827A CN 106587506 B CN106587506 B CN 106587506B
- Authority
- CN
- China
- Prior art keywords
- liquid
- parts
- solid
- adds
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 19
- 229910052785 arsenic Inorganic materials 0.000 title claims abstract description 15
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 title claims abstract description 15
- 239000003344 environmental pollutant Substances 0.000 title claims abstract description 10
- 231100000719 pollutant Toxicity 0.000 title claims abstract description 10
- 239000002351 wastewater Substances 0.000 title claims abstract description 7
- 239000007788 liquid Substances 0.000 claims abstract description 31
- 241000894006 Bacteria Species 0.000 claims abstract description 25
- 239000010865 sewage Substances 0.000 claims abstract description 25
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 8
- 239000002131 composite material Substances 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 8
- 238000006243 chemical reaction Methods 0.000 claims abstract description 5
- 239000013618 particulate matter Substances 0.000 claims abstract description 4
- 238000004062 sedimentation Methods 0.000 claims abstract description 4
- 238000000926 separation method Methods 0.000 claims abstract description 4
- 238000009288 screen filtration Methods 0.000 claims abstract 2
- 241000316848 Rhodococcus <scale insect> Species 0.000 claims description 14
- 108090000623 proteins and genes Proteins 0.000 claims description 14
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 9
- 230000001580 bacterial effect Effects 0.000 claims description 9
- 241000194103 Bacillus pumilus Species 0.000 claims description 6
- 241001509286 Thiobacillus denitrificans Species 0.000 claims description 6
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 150000001413 amino acids Chemical class 0.000 claims description 5
- 230000035772 mutation Effects 0.000 claims description 4
- 241000222393 Phanerochaete chrysosporium Species 0.000 claims description 3
- 240000007594 Oryza sativa Species 0.000 claims 1
- 235000007164 Oryza sativa Nutrition 0.000 claims 1
- 235000009566 rice Nutrition 0.000 claims 1
- CPJBEAMIONWDDK-UHFFFAOYSA-N [N].N.[P].[S] Chemical compound [N].N.[P].[S] CPJBEAMIONWDDK-UHFFFAOYSA-N 0.000 abstract description 2
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 239000000126 substance Substances 0.000 description 12
- 230000000694 effects Effects 0.000 description 10
- 238000011282 treatment Methods 0.000 description 8
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 6
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 6
- 241000589614 Pseudomonas stutzeri Species 0.000 description 5
- 238000010353 genetic engineering Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- 241000736131 Sphingomonas Species 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 239000002068 microbial inoculum Substances 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 3
- 235000017491 Bambusa tulda Nutrition 0.000 description 3
- 241001330002 Bambuseae Species 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 3
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 3
- 239000011425 bamboo Substances 0.000 description 3
- 239000003610 charcoal Substances 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229910001385 heavy metal Inorganic materials 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 230000000869 mutational effect Effects 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000003403 water pollutant Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 229920001661 Chitosan Polymers 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 101150110933 PHY gene Proteins 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 239000003124 biologic agent Substances 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000007403 mPCR Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000000053 physical method Methods 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000004065 wastewater treatment Methods 0.000 description 2
- 210000002268 wool Anatomy 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000187693 Rhodococcus rhodochrous Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241001135759 Sphingomonas sp. Species 0.000 description 1
- CVTZKFWZDBJAHE-UHFFFAOYSA-N [N].N Chemical class [N].N CVTZKFWZDBJAHE-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000007233 catalytic pyrolysis Methods 0.000 description 1
- 238000009388 chemical precipitation Methods 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000000909 electrodialysis Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 210000004209 hair Anatomy 0.000 description 1
- 239000010842 industrial wastewater Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000004630 mental health Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000009972 noncorrosive effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F9/00—Multistage treatment of water, waste water or sewage
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/001—Processes for the treatment of water whereby the filtration technique is of importance
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F2001/007—Processes including a sedimentation step
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/103—Arsenic compounds
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Mycology (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明涉及一种去除含砷污水污染物的方法，包括如下步骤:首先将污水经过固液分离器，进行固液分离，去除大块固体颗粒物质，随后液体进入沉淀池，沉淀12小时，再将液体通过圆孔过滤网去除固体絮凝物，经圆孔过滤网过滤的液体进入生物反应池，调节PH为7，按每立方米液体每次投加复合菌剂10克，每天投加1次，连续投加一周，最后静置3天，将液体排出。本发明的方法能够有效地去除含有氨氮硫磷以及砷的污染物，并且投入成本低，具备较好的应用前景。
Description
技术领域
本发明属于环保技术领域，具体涉及一种去除污水中污染物的方法。
背景技术
伴随着工业生产的发展以及人们生活水平的提高，工业污水量以及城市生活污水量正以惊人的速度猛增，这些污水或已经或正在污染着人类赖以生存的江、河及湖泊，已构成威协人类生存环境的原因之一。
为了满足公众对环境质量要求的不断提高，国家对氮制订了越来越严格的排放标准，研究开发经济、高效的除氮处理技术已成为水污染控制工程领域研究的重点和热点。虽然有许多方法都能有效地去除氨，如物理方法有反渗透、蒸馏、土壤灌溉；化学法有离子交换法、氨吹脱、化学沉淀法、折点氯化、电渗析、电化学处理、催化裂解；生物方法有硝化及藻类养殖，然而物理方法处理效果不佳，较之化学法，生物方法处理废水有如下优点:1)每种化学用品都是针对性很强的产品，当遇到其他化学物质时就有可能失效，而生物制剂对污染物的去除具有光谱性；2)化学产品可以暂时消除某些有害物质以及掩盖臭味，缺不能阻止有害物质的生成；3)使用化学产品后，水体中会有残留，可能导致二次污染。生物制剂所含天然微生物，不含致病菌和病原体，这些微生物在酶的催化作用下，以污水中的有机营养物质为食物，当污水得到净化后，这些微生物会随污染物的降低而逐渐减少，直至消亡；4)无毒，无腐蚀性，使用方便，基本不需要添加设备或是工程，节省资金投入。
污水主要有生活污水和工业废水。工业污水成分比较复杂，特别是大量的人工合成化合物进入环境，这类物质主要是氨氮类、硫化物以及含磷化合物，由于这些物质本身结构的复杂性，在短时间内不能被微生物分解利用，传统的废水处理方法用活性污泥培养驯化的微生物已不能有效地对这些污染物加以去除，这些物质长期在环境中积累，给我们赖以生存的生态环境造成很大污染，给人类的身心健康带来很大危害。我国相当一部分工业污染企业宁可受罚也不愿意投资治理废水，即使有污水处理装置运行也极不正常。因此，开发一种建设投资少、运行成本低、处理效率好的污水处理技术迫在眉睫。
发明内容
为了克服现有技术的不足，有效简单地去除污水中的氨氮硫磷以及重金属等污染物，本发明提供了一种去除污水中污染物的方法，其技术方案是通过如下方法实现的:
一种去除含砷污水污染物的方法，其包括如下步骤:
复合菌剂的制备:将混合菌液和载体按照1:1的重量比混合，搅拌均匀，然后静置6小时，最后置4℃下低温干燥，干燥后含水量控制在6％，即得；所述载体由竹炭、壳聚糖以及硅藻土按照2:2:1的质量比例混合得到，竹炭的粒径优选10目；
污水预处理:首先将污水经过固液分离器，进行固液分离，去除大块固体颗粒物质，随后液体进入沉淀池，沉淀12小时，再将液体通过圆孔过滤网去除固体絮凝物，圆孔过滤网的圆孔直径为0.1mm,生物氧化:经圆孔过滤网的液体进入生物反应池，调节pH为7，按每立方米液体每次投加复合菌剂10克，每天投加I次，连续投加一周，最后静置3天，将液体排出。
优选的，所述复合菌剂由如下重量份的原料菌混合而成:
红球菌10份，脱氮硫杆菌9份，施氏假单胞菌7份，鞘氨醇单孢菌6份，短小芽孢杆菌5份，黄孢原毛平革菌2份；上述各原料菌的浓度均控制在(1-2)X108个/ml。上述菌种可以是现有技术的常规菌株，
优选地，
所述红球菌为红球菌(Rhodococcusrhodochrous)ATCC15906；
所述脱氮硫杆菌为脱氮硫杆菌(Thiobacillus denitrificans)ATCC25259；
所述施氏假单胞菌为施氏假单胞菌(Pseudomonas stutzeri)CCTCC NO:M209107；
所述鞘氨醇单孢菌为鞘氨醇单孢菌(Sphingomonas sp.)CGMCC N0.4589；
所述短小芽孢杆菌为短小芽孢杆菌(Bacillus pumilus)ATCC27142；
所述黄抱原毛平革菌为黄抱原毛平革菌(Phanerochaete chrysosporium)ATCC24725。
本发明所述的菌均可以从CGMCC、CCTCC以及美国模式培养物集存库(ATCC)等商业途径购买得到。
优选的，所述红球菌为转入了编码如下任一个蛋白变体的编码基因的ATCC15906菌株,所述蛋白变体为相对于蛋白原始氨基酸序列分别进行如下突变62K/G、80E/V、93V/H、120R/I、131F/P、143Q/S、152P/Q、169G/I、179Q/A、192Q/G、241W/S、258R/T、263R/P,蛋白原始氨基酸序列参见GenBank：NP_745067.1。
本发明同时提供一种能够高效吸收砷的转基因菌株，该菌株通过将氨基酸序列如Genbank：NP_745067.1所示的序列导入到红球菌中实现其目的。
并且所述的氨基酸序列可以具有如下不用的位点的突变，62K/G、80E/V、93V/H、120R/I、131F/P、143Q/S、152P/Q、169G/I、179Q/A、192Q/G、241W/S、258R/T、263R/P。(62K/G表示在原始序列第62位的K氨基酸替换为G氨基酸)。这些突变后的氨基酸序列舍得导入了该突变后的氨基酸序列的菌株同样具有相似的效果。而申请人通过大量的实验证实，并不是所有的取代都具有相似的效果，有大量的取代之后的红球菌与其它菌株一起并不具有较好的污水处理的效果。
利用DNAMAN软件设计引物，分别加入了酶切位点，根据氨基酸序列NP_745067.1全基因合成所述的核苷酸序列PHY基因，通过进行扩增获得目的片段，PCR扩增获得目的基因PHY(同时通过多重PCR将相应的突变位点引入到基因序列中，从而获得了不同突变体基因)，用内切酶进行双酶切PCR产物，与同样经过双酶切的表达载体pEASY相连，将验证成功的重组质粒转化进入到所述红球菌中，获得降解砷的基因工程菌。
本发明的各菌种的扩大培养均为本领域的常规培养方式，也可以参照文献中记载的培养方式获得。
本发明取得的主要有益效果如下:本发明采用纯菌剂处理污水，有效地净化了污水，并且除磷、脱氮同时去除，在此基础上同时能够去除污水华中的重金属砷解决了传统工艺中除磷与脱氮效果难以同时兼顾重金属去除的矛盾，具有较好的应用前景。
具体实施方式
实施例1
一种去除污水中污染物的方法，其包括如下步骤:
复合菌剂的制备:将混合菌液和载体按照1:1的重量比混合，搅拌均匀，然后静置6小时，最后置4℃下低温干燥，干燥后含水量控制在6％，即得；载体由竹炭、壳聚糖以及硅藻土按照2:2:1的质量比例混合得到；上述混合菌液由如下重量份的原料菌混合而成:红球菌10份，脱氮硫杆菌9份，施氏假单胞菌7份，鞘氨醇单孢菌6份，短小芽孢杆菌5份，黄孢原毛平革菌2份；上述各原料菌的浓度均控制在1×108个/ml。
污水预处理:首先将污水NH3-N为300mg/L，硫化物80mg/L，含磷污染物为70mg/L，砷含量为20mg/ml，经过固液分离器，进行固液分离，去除大块固体颗粒物质，随后液体进入沉淀池，沉淀12小时，再将液体通过圆孔过滤网去除固体絮凝物，圆孔过滤网的圆孔直径为0.1mm,
生物氧化:经圆孔过滤网的液体进入生物反应池，调节pH为7.0，按每立方米液体每次投加复合菌剂10克，每天投加1次，连续投加一周，最后静置3天，将液体排出。经检测，污水中氨氮的含量分别为12.5mg/L，硫化物4.5mg/L，含磷污染物为2.5mg/L，去除率均达到96％以上；而砷含量为17mg/mL，去除效果不显著。
实施例2
利用DNAMAN软件设计引物，分别加入了酶切位点，根据氨基酸序列NP_745067.1全基因合成所述的核苷酸序列PHY基因，通过进行扩增获得目的片段，PCR扩增获得目的基因PHY(同时通过多重PCR将相应的突变位点62K/G、80E/V、93V/H、120R/I、131F/P、143Q/S、152P/Q、169G/I、179Q/A、192Q/G、241W/S、258R/T、263R/P引入到基因序列中，从而获得了不同突变体基因)，用加入的酶切位点进行双酶切PCR产物，与同样经过双酶切的克隆表达载体pEASY相连，将验证成功的重组质粒转化进入到所述红球菌ATCC15906菌株中，获得降解砷的基因工程菌。
实施例3红球菌基因工程菌与其余几种菌剂的污水处理效果验证
按照实施例1的方法，进行相应的污水处理实验，其污水与实施例1的污水属于同一批次，具有相同浓度的污染物浓度。其中菌剂的各组分组成与实施例1的完全相同。
分别采用实施例2所述的不同的突变位点的基因工程菌进行去除砷的反应，通过实验发现，62K/G、80E/V、93V/H、120R/I、131F/P、143Q/S、152P/Q、169G/I、179Q/A、192Q/G、241W/S、258R/T、263R/P这几种突变菌都具有相对于原始菌具有明显增强的效果。总处理时间不大于4天，其结果如下：污水为同一批污水，从而保证条件一致。
污水的各污染物浓度：NH3-N为300mg/L，硫化物80mg/L，含磷污染物为70mg/L，砷含量为20mg/ml。
从以上结果可以看出，采用基因工程构建的红球菌与其它几种菌株构成的菌剂除了具有原始较好的去除氨氮、硫化物以及磷的效果之外，还具有较好的去除砷的效果，而且在去除氨氮、硫化物以及磷方面基因工程的红球菌与其它几种菌株之间还具有增强的协同作用。
Claims (2)
1.一种去除含砷污水污染物的方法，其包括如下步骤:首先将污水经过固液分离器，进行固液分离，去除大块固体颗粒物质，随后液体进入沉淀池，沉淀12小时，再将液体通过圆孔过滤网去除固体絮凝物，经圆孔过滤网过滤的液体进入生物反应池，调节pH为7，按每立方米液体每次投加复合菌剂10克，每天投加I次，连续投加一周，最后静置3天，将液体排出，所述的复合菌剂由如下重量份的原料菌混合而成:红球菌10份，脱氮硫杆菌9份，施氏假单胞菌7份，鞘氨醇单胞菌6份，短小芽孢杆菌5份，黄孢原毛平革菌2份；上述各原料菌的浓度均控制在(1-2)X108个/ml，所述红球菌为转入了编码如下蛋白变体的编码基因的ATCC15906菌株,所述蛋白变体为相对于蛋白原始氨基酸序列进行如下突变:80E/V,蛋白原始氨基酸序列参见GenBank：NP_745067.1。
2.一种蛋白变体，其特征在于：所述蛋白变体为相对于蛋白原始氨基酸序列进行如下突变:80E/V,蛋白原始氨基酸序列参见GenBank：NP_745067.1。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611187827.3A CN106587506B (zh) | 2015-07-26 | 2015-07-26 | 一种去除含砷污水污染物的方法 |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510441623.7A CN105060620B (zh) | 2015-07-26 | 2015-07-26 | 一种去除含砷污水污染物的方法 |
| CN201611187827.3A CN106587506B (zh) | 2015-07-26 | 2015-07-26 | 一种去除含砷污水污染物的方法 |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510441623.7A Division CN105060620B (zh) | 2015-07-26 | 2015-07-26 | 一种去除含砷污水污染物的方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106587506A CN106587506A (zh) | 2017-04-26 |
| CN106587506B true CN106587506B (zh) | 2019-07-05 |
Family
ID=54490210
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510441623.7A Active CN105060620B (zh) | 2015-07-26 | 2015-07-26 | 一种去除含砷污水污染物的方法 |
| CN201611187827.3A Active CN106587506B (zh) | 2015-07-26 | 2015-07-26 | 一种去除含砷污水污染物的方法 |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510441623.7A Active CN105060620B (zh) | 2015-07-26 | 2015-07-26 | 一种去除含砷污水污染物的方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN105060620B (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09314185A (ja) * | 1996-05-30 | 1997-12-09 | Ebara Corp | 有機性汚水の処理方法及び処理装置 |
| CN101050030A (zh) * | 2007-03-21 | 2007-10-10 | 李昕 | 一种复合功能菌处理危险废物砷废液的方法 |
| CN103449680A (zh) * | 2013-09-13 | 2013-12-18 | 李丽萍 | 一种去除污水中污染物的方法 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103449681B (zh) * | 2013-09-13 | 2014-11-26 | 何洋 | 一种利用微生物制剂处理污水的方法 |
-
2015
- 2015-07-26 CN CN201510441623.7A patent/CN105060620B/zh active Active
- 2015-07-26 CN CN201611187827.3A patent/CN106587506B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09314185A (ja) * | 1996-05-30 | 1997-12-09 | Ebara Corp | 有機性汚水の処理方法及び処理装置 |
| CN101050030A (zh) * | 2007-03-21 | 2007-10-10 | 李昕 | 一种复合功能菌处理危险废物砷废液的方法 |
| CN103449680A (zh) * | 2013-09-13 | 2013-12-18 | 李丽萍 | 一种去除污水中污染物的方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105060620B (zh) | 2017-05-31 |
| CN106587506A (zh) | 2017-04-26 |
| CN105060620A (zh) | 2015-11-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105036471B (zh) | 一种去除含铅污水污染物的方法 | |
| CN105130100B (zh) | 一种去除含镉污水污染物的方法 | |
| CN106007243B (zh) | 一种去除含铬污水污染物的方法 | |
| CN106430865B (zh) | 一种去除含汞污水污染物的方法 | |
| CN106587506B (zh) | 一种去除含砷污水污染物的方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20190610 Address after: 432000 Three Li village, Changjiang port office, Yingcheng City, Xiaogan, Hubei Applicant after: Hubei Kang Chuang Technology Co., Ltd. Address before: 100193 China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing Applicant before: Li Na |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |