CN105858977B - 一种处理污水中农药残留和重金属汞的方法 - Google Patents
一种处理污水中农药残留和重金属汞的方法 Download PDFInfo
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Abstract
本发明涉及利用污水处理领域，具体地说是一种降低污水中汞和农药残留的污染的方法。该方法能够有效去除污水中汞污染具有效率高、营养要求低、操作简单及安全性高等优点，与现有的其他工艺相比，显著降低了污水汞和农药残留污染治理的成本，提高了污水中汞和农药污染治理的效率，具有非常重要的实际应用价值。
Description
本申请是申请日为2015年6月21日、申请号为201510343958.5、发明名称为“一种处理污水中农药残留和重金属汞的方法”的发明专利申请的分案申请。
技术领域
本发明属于污水处理领域，具体涉及污水中农药残留和重金属汞的处理，属于污水综合处理技术领域。
背景技术
近几十年来，化学农药一直是园林农业中控制病虫草、鼠害的主要手段品种和销量逐年增加。化学农药在控制病虫草、鼠害提高产量的同时，也破坏了生态，污染了环境，每年50万余吨的农药用量的70％将流入江河土壤，而且有的农药消解速度慢，空气中农药浓度将维持一个相当长的时间，并由于其残留对水域、食品的污染直接危害了人体健康以及动物的安全，污染的地表水又直接污染着地下水。而为确保粮食丰收，农药又是必不可少的，因此，脱毒、净化水资源，综合治理水环境污染刻不容缓，势在必行。
农药污染对人类健康亦造成极大危害。据估计，全世界每年有两百万人农药中毒，两万人死亡。Pimentel等认为农药的使用给美国造成的直接损失每年达30亿美元，引起社会与环境的总代价高达81亿美元，相当于美国化学农药效益值162亿美元的50％。所以寻找处理环境中农药污染的方法成为重要研究课题。
另外，随着我国工业化进程的不断加快，环境污染也不断加剧，治理人类赖以生存的土壤及大气污染已经迫在眉睫。除了严格控制污染物的排放外，研究开发适合我国国情的环境污染治理技术具有重要的意义。环境污染包括重金属污染、农药和持久性有机化合物污染、化肥施用污染等。随着人口增加及经济发展，大量未经处理的废弃物向农田转移，我国面临的土壤及水体污染问题越来越严重。近年来我国的土壤污染正在向不同尺度的区域性发展，并对各种农产品品质产生严重影响。据不完全统计，我国重金属污染的土壤面积达2000万公顷，占总耕地面积的1/6。因工业“三废”污染的农田近700万公顷，使粮食每年减产100亿公斤。在一些地区汞、砷、铜、锌等元素的超标面积占污染总面积的一半左右。一些主要蔬菜基地土壤镉污染普遍，其中有的市郊大型设施蔬菜园艺场中，土壤中锌含量高达517毫克/千克，超标5倍之多。由于土壤污染具有隐蔽性，潜伏性和长期性，其严重后果仅能通过食物给动物和人类健康造成危害，因而不易被人们察觉。特别是土壤中的重金属具有流动性小，污染时间长，通过物理或化学方法很难彻底清除等特点。改善生态环境，保护土壤质量，控制与修复土壤污染，才能实现农业安全，这是我国目前面临的一个严峻而又紧迫的重大问题。
目前，农药污染以及重金属污染的处理方法很多，概括起来包括化学处理、焚烧、掩埋和生物整治等方法。化学处理方法可行性强，但在处理过程中产生大量的酸和碱，这些物质还必须再加处理。焚烧可能是降解这些污染物最可靠的方法。但是，处理过程中放出大量的毒气，而且需要大量的热能。掩埋也存在问题，主要表现在对其周围的土壤和水源造成二次污染。因此，开发一种节能环保成本低的处理方法迫在眉睫。
发明内容
本发明的目的之一在于提供一种处理污水中农药残留和重金属汞的方法。
本发明是采用如下技术方案实现的:
一种处理污水中农药残留和重金属汞的方法，包括如下步骤:
(1)预处理:通过过滤和沉淀，去除所处理污水中的悬浮物；
(2)主处理:污水部分根据水中汞浓度加入药剂硫化铵、助凝剂(聚丙烯酰胺)和絮凝剂(聚合氯化铝),加入到污水中，于常温条件下，流动培养24小时，即可清除污水中汞；
(3)后处理:将污水再次经过沸石和活性炭吸附池，吸附农药残留；
(4)将处理后的污水经消毒处理后，直接排放或回用。
本发明提供的另一个技术方案是:
一种处理污水中农药残留和重金属汞的方法，包括如下步骤:
(1)预处理:通过过滤和沉淀，去除所处理污水中的悬浮物；
(2)主处理:将培养至对数生长期的细菌恶臭假单胞菌(Pseudomonas putida),加入到污水中，于常温条件下，流动培养24小时，即可清除污水中汞；
(3)后处理:将污水再次经过沸石和活性炭吸附池，吸附农药残留；
(4)将处理后的污水经消毒处理后，直接排放或回用。
优选的，所述的恶臭假单胞菌为转入了经修饰的细胞色素氧化还原酶(P450CYP9G2)和NADPH-细胞色素P450还原酶(POR)的基因的Pseudomonas putida SPlCGMCC N0.3887菌株，所述的NADPH-细胞色素P450还原酶(POR)的基因所编码的蛋白序列参见GenBank：L19897，所述的经修饰的细胞色素氧化还原酶(P450CYP9G2)基因所编码的蛋白序列为相对于细胞色素氧化还原酶(P450CYP9G2)原始氨基酸序列分别进行如下突变Ser30Ile(表示第30位的Ser突变为Ile)，Phe56Gly，Gly77Lys，Phe186Ala，Leu276Ile，Leu311Asp，Val366Gly，Val409Cys，Asp415Pro，Trp426Leu，Lys476Gly，细胞色素氧化还原酶(P450CYP9G2)基因所编码的原始蛋白序列参见GenBank：AB096739。
本发明提供的一系列经过基因改造的菌株不仅具有较好的去除汞的效果，同时还能够有效的去除污水中的农药残留，具有较好的应用前景。
上述方法具有以下有益效果:
1、通过本发明的处理方法，大大提高了汞和农药残留的处理效果，降低了反应时间。
2、提供一种降解汞和农药残留的转化菌株，具有较高的去除效率。
为了便于理解，以下将通过具体的实施例对本发明进行详细地描述。需要特别指出的是，具体实例仅是为了说明，并不构成对本发明范围的限制。显然本领域的普通技术人员可以根据本文说明，在本发明的范围内对本发明做出各种各样的修正和改变，这些修正和改变也纳入本发明的范围内。
具体实施方式
实施例1汞和农药残留污水的处理
污水进水时汞的浓度为30mg/L，乐果20mg/L，杀螟硫磷20mg/L，灭多威20mg/L。
预处理:通过过滤和沉淀，去除所处理污水中的悬浮物；
主处理:将培养至对数生长期的细菌恶臭假单胞菌Pseudomonas putida SPlCGMCC N0.3887(来自CN 102373161A),加入到污水中，于常温条件下，流动培养24小时，即可清除污水中汞；
后处理:将污水再次经过沸石和活性炭吸附池过夜处理，吸附农药残留；
将处理后的污水经消毒处理后，直接排放或回用。经检测，汞的浓度为2.2mg/L，乐果15.3mg/L，杀螟硫磷14.4mg/L，灭多威16.5mg/L。
实施例2降解农药基因的改造与获得
按照CN1970741A中公开的方法，克隆获得CYP9G2和POR基因。针对CYP9G2基因进行了如下改造，在GenBank：AB096739的基础上，通过多重PCR将其编码基因相应的Ser30Ile(表示第30位的Ser突变为Ile)，Phe56Gly，Gly77Lys，Phe186Ala，Leu276Ile，Leu311Asp，Val366Gly，Val409Cys，Asp415Pro，Trp426Leu，Lys476Gly，Leu500Ala，突变位点分别引入到GenBank：AB096739所示蛋白中，从而构建了不同的突变体CYP9G2基因(可参考现有技术的制备方法即可获得)。将突变后的CYP9G2基因与POR基因一起按照制备CN1970741A的方法分别于表达载体相连，将验证成功的重组质粒转化进入Pseudomonas putida SPl CGMCCN0.3887菌株中，通过PCR检测，其构建成功。
实施例3污水降解效果的验证
将含有不同突变蛋白的菌株，分别都按照实施例1的方法，验证对污水的处理效果。污水进水时汞的浓度为30mg/L，乐果20mg/L，杀螟硫磷20mg/L，灭多威20mg/L。以转入未突变CYP9G2和POR基因的菌株和的菌作为对照。具体结果如下：
从以上的结果可以看出，导入了特定突变的CYP9G2和POR基因的菌具有增强的去除汞和农药的效果，这充分说明并不是任意的改变都具有较好的效果。申请人通过大量的验证才获得了具有较好效果的几个位点。
Claims (2)
1.一种处理污水中农药残留和重金属汞的方法，包括如下步骤:
(1)预处理:通过过滤和沉淀，去除所处理污水中的悬浮物；
(2)主处理:将培养至对数生长期的细菌恶臭假单胞菌(Pseudomonas putida),加入到污水中，于常温条件下，流动培养24小时，即可清除污水中汞；
(3)后处理:将污水再次经过沸石和活性炭吸附池，吸附农药残留；
(4)将处理后的污水经消毒处理后，直接排放或回用；
所述的恶臭假单胞菌为转入了经修饰的细胞色素氧化还原酶和NADPH-细胞色素P450还原酶的基因的Pseudomonas putida SPl CGMCC N0.3887菌株，所述的NADPH-细胞色素P450还原酶的基因所编码的蛋白序列参见GenBank：L19897，所述的经修饰的细胞色素氧化还原酶基因所编码的蛋白序列为相对于细胞色素氧化还原酶原始氨基酸序列进行如下突变Val366Gly，细胞色素氧化还原酶基因所编码的原始蛋白序列参见GenBank：AB096739。
2.一种蛋白变体，其特征在于：所述的变体为相对于蛋白原始氨基酸序列进行如下突变Val366Gly，原始蛋白序列参见GenBank：AB096739。
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