Difference between revisions of "Team:UESTC-China/description"

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<a href="#TCP-Pollution">1,2,3-TCP Pollution</a>
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<a href="#Harm">Harm of 1,2,3-TCP</a>
 
</li>
 
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<li>
 
<li>
<a href="#Phytodegradation">Phytodegradation</a>
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<a href="#Technology">Technology to treat</a>
 
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<a href="#Our-Pathway">Our Pathway</a>
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<a href="#Strategy">Our strategy</a>
 
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<div id="Overview">
 
<div id="Overview">
 
<h2>Overview</h2>
 
<h2>Overview</h2>
<br /><p>化学工业的进步给我们的生活带来了巨大的便利,许多功能近乎于“魔法”的产品纷纷被生产出来。但是,化学工业为我们带来便利的同时,无形中也让我们付出了巨大的代价。大量的化学污染物通过各种途径被排放出来,污染我们的环境。其中,有机氯化在这些污染物中占有很大一部分的比例。我们队伍今年的项目是针对1,2,3 三氯丙烷——一种不太受关注但威胁越来越重的一种人工合成的有机氯化物。我们打算通过往烟草中转入一种三酶体系的质粒来有效降解1,2,3-三氯丙烷。</p>
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<br /><p>Chemical industry brings us comfortable life, but it also let us pay a heavy price at the same time. A large number of chemical contaminants are discharge through various means, polluting our environment. And organic halide is important part in these pollution. Our project in this year is directed against 1,2,3-Trichloropropane – an organic chloride which is a less concerned but unsecured pollution. We intend to effectively degrade 1,2,3-trichloropropane by transferring it into a three-enzyme plasmid to the Nicotiana tabacum.</p>
 
</div>
 
</div>
  
<div id="TCP-Pollution">
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<div id="Harm">
<h2>The 1,2,3- TCP pollution</h2>
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<h2>Harm of 1,2,3-TCP</h2>
<br /><p>1,2,3-tcp是一种新兴的有机污染物,它的化学性质稳定并且具有突变、致病、致癌等毒害作用。物理性质上则表现为不易容易溶于水,且密度比水大。在工业上经常单独用作工业溶剂、脱脂剂、脱漆剂等,同时也作为二氯丙烯、环氧氯丙烷等化工厂产品的中间体或者副产物出现。其最早的污染记载是1940s作为陶氏化学和壳牌公司生产的土壤蒸熏剂的成分被播撒到加州各地的农田中</p>
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<br /><p>1,2,3-Trichloropropane(1,2,3-TCP), an emergent organic pollutant, formed as a by-product during the synthesis of various chemicals and was present in commercial preparations of the soil fumigant 1,3-dichloropropene (also known under the trade name D-D).</p>
<br /><p class="pic"><img src="https://static.igem.org/mediawiki/2017/3/34/T--UESTC-China--introduction_1.png" style="width: 40%;"/></p>
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<br /><p class="pic"><img src="https://static.igem.org/mediawiki/2017/5/5d/T--UESTC-China--description_1.png" style="width: 50%;"/></p>
<br /><p>自1970s以来陆续在美国、日本等发达国家的饮用水中检测到该成分,但由于一直未确定其致癌性且含量不高所以一直未受到重视。但近10年来各地饮用水中检测到tcp的次数增加,其毒性和致癌性的关注度开始提升,美国环保署和国防部视其为“新兴污染物”。Tcp微溶于水,且密度比水大。这意味其一旦进入环境中便会不断在土壤中下渗并且很可能进入地下水中。曾有人在中国河北省一座70年代短暂运行过的化工厂附近进行过土壤调查,采集了大约5~15米处的土壤和地下水样品。在这些样品中都发现了浓度不低的tcp,最高浓度甚至可以达到3890mg/KG。这一方面表现了1,2,3-TCP 的稳定性,可以在自然界中存在四十而不降解。另一方面,深入地下10几米也体现了其密度较大、易渗透的特性。该调查和相关其他研究都证明了tcp在底下环境难以自然降解,可以在土壤和地下水中存在数十年。</p>
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<p class="mid">Figure 1  The earliest record about 1,2,3-TCP is that it was spread to the fields around California</p>
<br /><p style="color: red;">这种现象是非常值得引起重视的,它稳定、易渗透的特性使得其一旦不被及时处理,便会慢慢渗入土壤深处,使其变得难以处理。</p>
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<p class="mid">as a part of soil fumigants(D-D and Telone) produced by Dow Chemical and Shell.</p>
<br /><p class="pic"><img src="https://static.igem.org/mediawiki/2017/0/01/T--UESTC-China--introduction_2.jpg" style="width: 60%;"/></p>
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<br /><p>Having detected frequently in various occasions in recent years, 1,2,3-TCP has gradually aroused people's attention in the world. Animal experiments show that it has a great deal of damage to livers and kidneys of the animals and makes them die at certain concentration. It has been classified as "likely to be carcinogenic to humans" by the EPA. Dr. Qian Yong the concentration up to 3890mg / L of 1,2,3-TCP from 5m to 15m underground at the ruins of a factory, running in the 1970s shows.</p>
<br /><p style="color: red;">并且tcp有高的迁移率,接近地下水的迁移速度。这代表着tcp极易扩散进入我们的生活区域和生活用水中。所以处理排放到自然界的1,2,3-tcp的最佳时机就是在其排放时间不是很久,还停留土壤表面或者浅层时就开始处理。</p>
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<br /><p class="pic"><img src="https://static.igem.org/mediawiki/2017/c/cf/T--UESTC-China--description_2.png" style="width: 50%;"/></p>
<br /><p>但遗憾的是,世界各地对它的重视还远远不够,在美国只有夏威夷、加州等地出台了相关的排放标准。而在中国,tcp甚至没有被列入水质检测物质的名单中!<span style="color: red;">也正是了解到这个因素,我们队伍一方面设计了这种带有三酶体系的烟草来降解土壤中的1,2,3-tcp,从技术思考解决问题的方案。</span>另一方面希望通过通过自身的宣传和iGEM比赛的影响力引起社会的重视,在tcp还没对社会造成更大危害之前及时遏制事态的发展!</p>
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<p class="mid">Figure 2 1,2,3-TCP flowed to the depths of the ground and kept about 30 years,</p>
 +
<p class="mid">which make it easy enter into people’s life with groundwater.</p>
 +
 +
<br /><p>The discovery proves the stability and permeability of 1,2,3-TCP. By the way, some researches show the adhesion of 1,2,3-TCP is very low, which means it easy to spread in nature and enters to people's life through a variety of ways such as groundwater, threatening people's health. The tap water around the world has been detected 1,2,3-TCP frequently, proving the harmfulness of 1,2,3-TCP.</p>
 +
<p class="pic"><img src="https://static.igem.org/mediawiki/2017/4/4b/T--UESTC-China--description_3.png" style="width: 90%;"/></p>
 +
<p class="mid">Figure 3  The 1,2,3-TCP was frequently detected in the recent years</p>
 +
<br /><p>However, governments don't pay enough attention to 1,2,3-TCP. Some counties, like China and American, even don't include 1,2,3-TCP into the pollutant detection list. Under these circumstances, we hope that we can attract the attention of the society and contain the spread of 1,2,3-TCP pollution through this project.</p>
 
</div>
 
</div>
  
<div id="Phytodegradation">
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<div id="Technology">
<h2>Why does we choose Phytodegradation</h2>
+
<h2>Technology to treat 1,2,3-TCP</h2>
<br /><p>目前处理1,2,3-tcp 的主要常规方法有 活性炭吸附法、hydrogen release compound (HRC)、零价锌三种。但是活性炭吸附法吸收效率较低且作用缓慢。而HRC、零价锌等方法价格比较昂贵,且所需条件较多,不适合投入到环境复杂的自然界中,目前只是在用于实验室环境中。</p>
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<br /><p>Traditional remediation technology to treat 1,2,3-TCP includes granular activated carbon (GAC), dechlorination by hydrogen release compound (HRC®), reductive dechlorination by zero valent zinc and others. They have difference disadvantages such as inefficient, expensive or impractical. So it is hard to treat 1,2,3-TCP in large-scale. In recent years, the concept of "Bioremediation" has been respected by people. There is someone that has successfully introduced the genes of specific enzyme which can degrade 1,2,3-TCP into microorganism such as Escherichia coli, Pseudomonas putida. These strain are efficient. However, it’s shortcomings are also very obvious. First, the strain usually work in the bioreactor with favourable bacterial density and continuous nutrient input. It is very difficult and expensive to make it in nature. Second, some strain like Pseudomonas putida is a kind of pathogen and them may be create security issues. At last, the degradation of them will produce secondary pollution. On one word, the “Bioremediation” to 1,2,3-TCP isn’t a safe and efficient strategy for us now. So, we paid our attention to "phytoremediation", which hasn’t been tried.</p>
<br /><p>所以在常规理化方法难以使用的情况下,利用生物1,2,3 tcp 的方法开始受到重视。作为最近社会上越来越受重视的“绿色修复”生物降解污染物(尤其是在天然条件下)越来越被人们所推崇。由于一些早期研究表明 1,2,3 tcp 可能在以O2为电子受体的氧化共代谢作用下通过生物作用转化为C〇2、H20和HC1,所以人们注意力主要集中在好氧微生物中。但是到目前为止,对能降解1,2,3 tcp的好氧微生物的富集和筛选都失败。反而在厌氧微生物中发现几个菌种可以在绝对厌氧的环境中可降解tcp,但是由于条件比较严苛,所以没能获得其转化的效率。而在最近几年有研究开始使用基因工程的方法获取能够降解tcp的微生物。如samin在恶臭假单胞菌中引入烷基脱氢霉素,对tcp的降解效率达到95%之高,可以说降解效率非常高。但是它的缺点也非常明显。首先,微生物降解污染常常是通过制作相关的生物反应器,这往往需要保持较大的菌群密度和连续的营养输入。而在实际使用中往往面对复杂的环境和当地菌群的竞争,在该种情况下完成这些目前耗费往往不小。其次,恶臭假单胞菌是一种致病菌,可能会导致人类和鱼类染病。再者它的降解终产物为会对环境产生二次污染的氯化物。所以通过微生物降解tcp以改善环境的行为在现有条件下是难以实现的。</p>
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<br /><p>基于以上考虑,我们决定使用植物作为载体来降解tcp。这个决定也正好与现下推崇的“植物修复”(Phytoremediation)的概念。植物修复比微生物通常生物修复的优点是作为具有大生物量的自养系统的植物只需要适度的营养输入就可以持久有效的处理污染物,应用和简单管理相对便宜。并且植物侵入性较差,在清理环境的同时还可以碳封存和土壤稳定以提供美观环境和野生动物栖息地。再者,用于植物修复的植物材料可以再加工成木片,纸浆,甚至某些对某些处理重金属的植物进行处理,将重金属回收。</p>
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<br /><p>“植物修复”主要是有四种方式:植物萃取、植物挥发、植物固定和植物降解。</p>
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<br /><p class="pic"><img src="https://static.igem.org/mediawiki/2017/f/f4/T--UESTC-China--introduction_3.jpg" style="width: 85%;"/></p>
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<br /><p>通过tcp理化性质的了解,我们的得知tcp通常不能在生物中富集,且其吸附力非常低易于迁移,所以难以通过萃取和固定的方式进行处理。况且如果采用植物萃取和植物固定的方式,就要经常对植物进行处理和更换。这些都是需要建立一套完整的后续处理体系,耗时耗力。而植物挥发就更不在考虑的范围内了,tcp本身便可以通过吸入的方式对人类造成严重伤害,所以这种方式根本不可能投入自然界中使用。由此看来,最后可选的方式便只有植物降解这一种了。</p>
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</div>
 
</div>
  
<div id="Our-Pathway">
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<div id="Strategy">
<h2>Our pathway to degradation 1,2,3-TCP</h2>
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<h2>Our strategy</h2>
<br /><p>选定采用植物降解这一策略后,我们便开始查找可用的降解TCP的通路。最后我们发现了Dvorak 等人在2014提出的通过固定化 氯代烷烃脱氯素酶(突变体)、氯代醇脱氯素酶和环氧化物酶(DhA31、HheC、EchA)三种酶降解tcp。</p>
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<br /><p>Compared to "Bioremediation", the advantage is very obvious. The most important one is that plant has a set of photosynthetic autotrophic system which means it can degrade 1,2,3-TCP in a long time and just need little nutrition input, easier and cheaper. The plant can aslo stabilized soil and absorb CO2 while clean the environment. Furthermore, plant material can be reworks into wood chips, pulp and some of the plants that treat heavy metals, and heavy metals are recovered.  "Phytoremediation" mainly contains four ways: Phytoextraction, Phytostabilization, Phytovolatilization, Phytodegradation.</p>
<br /><img src="https://static.igem.org/mediawiki/2017/0/0d/T--UESTC-China--introduction_4.png" style="width: 50%;"/>
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<p class="pic"><img src="https://static.igem.org/mediawiki/2017/0/0e/T--UESTC-China--description_4.png" style="width: 80%;"/></p>
<img src="https://static.igem.org/mediawiki/2017/e/ed/T--UESTC-China--introduction_5.png" style="width: 40%;"/>
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<p class="mid">Figure 4  The four main models of Phytoremediation</p>
<br /><p>该体系将tcp最终转化对环境无害的甘油,其最终转化效率为78%,转化为中间体的效率可以到到98%。这是我们目前能找到的降解tcp且终产物无害的最高效率,因此我们决定将该途径通过基因工程的方式转入到模式植物烟草中。在烟草中搭建体系主要是考虑到其生长就有强大的积极性。并且烟草作为一种模式生物,对基因表达和生长调控方面有相当数量的研究,具有一套成熟的遗传转化以及培养体系。因为这几种酶全部来源于原核生物中,没有在植物中发现过。在一个成熟的模式植物中进行尝试可以更加容易发现问题并且进行改进,也为以后我们在其他植株中进行尝试提供数据与方法。而谈到实际的使用,考虑到烟草生长的节律性等因素。我们可以在工厂的排放地以及一些污染较严重的地方集中种植并且对其生长条件进行一定的调控,形成规模效应以实现对1,2,3-tcp 的快速、大量的降解。对比需要完整的后续体系和投入的微生物修复技术,这样的方式无疑是持久、长效并且廉价的。</p>
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<br /><p>Combined with the physical and chemical properties of 1,2,3-TCP, we learn about that 1,2,3-TCP is unlikely to become concentrated in plants other aquatic organisms because it has a low estimated bioconcentration factor (BCF) and  sticking coefficient(lgKoc). So it is difficult to treat 1,2,3-TCP using phytoextraction and phytostabilization. Moreover, it needs a complex system to deal with these plant if we choose phytoextraction and phytostabilization, which require much time and effort. By the way, phytovolatilization is more unsuitable because 1,2,3-TCP , inhaled by human body, will create more damage in the gas. Thus, we finally identified the strategy of phytodegradation. We introduce the gene of three enzyme-haloalkane dehalogenase(dhA31),haloalcohol dehalogenase(HheC) Epichlorohydrin epoxide hydrolase (echA) into model plant- Nicotiana tabacum and transform 1,2,3-TCP into glycerol.</p>
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Revision as of 04:14, 12 October 2017

Team:UESTC-China/Introduction

Overview


Chemical industry brings us comfortable life, but it also let us pay a heavy price at the same time. A large number of chemical contaminants are discharge through various means, polluting our environment. And organic halide is important part in these pollution. Our project in this year is directed against 1,2,3-Trichloropropane – an organic chloride which is a less concerned but unsecured pollution. We intend to effectively degrade 1,2,3-trichloropropane by transferring it into a three-enzyme plasmid to the Nicotiana tabacum.

Harm of 1,2,3-TCP


1,2,3-Trichloropropane(1,2,3-TCP), an emergent organic pollutant, formed as a by-product during the synthesis of various chemicals and was present in commercial preparations of the soil fumigant 1,3-dichloropropene (also known under the trade name D-D).


Figure 1 The earliest record about 1,2,3-TCP is that it was spread to the fields around California

as a part of soil fumigants(D-D and Telone) produced by Dow Chemical and Shell.


Having detected frequently in various occasions in recent years, 1,2,3-TCP has gradually aroused people's attention in the world. Animal experiments show that it has a great deal of damage to livers and kidneys of the animals and makes them die at certain concentration. It has been classified as "likely to be carcinogenic to humans" by the EPA. Dr. Qian Yong the concentration up to 3890mg / L of 1,2,3-TCP from 5m to 15m underground at the ruins of a factory, running in the 1970s shows.


Figure 2 1,2,3-TCP flowed to the depths of the ground and kept about 30 years,

which make it easy enter into people’s life with groundwater.


The discovery proves the stability and permeability of 1,2,3-TCP. By the way, some researches show the adhesion of 1,2,3-TCP is very low, which means it easy to spread in nature and enters to people's life through a variety of ways such as groundwater, threatening people's health. The tap water around the world has been detected 1,2,3-TCP frequently, proving the harmfulness of 1,2,3-TCP.

Figure 3 The 1,2,3-TCP was frequently detected in the recent years


However, governments don't pay enough attention to 1,2,3-TCP. Some counties, like China and American, even don't include 1,2,3-TCP into the pollutant detection list. Under these circumstances, we hope that we can attract the attention of the society and contain the spread of 1,2,3-TCP pollution through this project.

Technology to treat 1,2,3-TCP


Traditional remediation technology to treat 1,2,3-TCP includes granular activated carbon (GAC), dechlorination by hydrogen release compound (HRC®), reductive dechlorination by zero valent zinc and others. They have difference disadvantages such as inefficient, expensive or impractical. So it is hard to treat 1,2,3-TCP in large-scale. In recent years, the concept of "Bioremediation" has been respected by people. There is someone that has successfully introduced the genes of specific enzyme which can degrade 1,2,3-TCP into microorganism such as Escherichia coli, Pseudomonas putida. These strain are efficient. However, it’s shortcomings are also very obvious. First, the strain usually work in the bioreactor with favourable bacterial density and continuous nutrient input. It is very difficult and expensive to make it in nature. Second, some strain like Pseudomonas putida is a kind of pathogen and them may be create security issues. At last, the degradation of them will produce secondary pollution. On one word, the “Bioremediation” to 1,2,3-TCP isn’t a safe and efficient strategy for us now. So, we paid our attention to "phytoremediation", which hasn’t been tried.

Our strategy


Compared to "Bioremediation", the advantage is very obvious. The most important one is that plant has a set of photosynthetic autotrophic system which means it can degrade 1,2,3-TCP in a long time and just need little nutrition input, easier and cheaper. The plant can aslo stabilized soil and absorb CO2 while clean the environment. Furthermore, plant material can be reworks into wood chips, pulp and some of the plants that treat heavy metals, and heavy metals are recovered. "Phytoremediation" mainly contains four ways: Phytoextraction, Phytostabilization, Phytovolatilization, Phytodegradation.

Figure 4 The four main models of Phytoremediation


Combined with the physical and chemical properties of 1,2,3-TCP, we learn about that 1,2,3-TCP is unlikely to become concentrated in plants other aquatic organisms because it has a low estimated bioconcentration factor (BCF) and sticking coefficient(lgKoc). So it is difficult to treat 1,2,3-TCP using phytoextraction and phytostabilization. Moreover, it needs a complex system to deal with these plant if we choose phytoextraction and phytostabilization, which require much time and effort. By the way, phytovolatilization is more unsuitable because 1,2,3-TCP , inhaled by human body, will create more damage in the gas. Thus, we finally identified the strategy of phytodegradation. We introduce the gene of three enzyme-haloalkane dehalogenase(dhA31),haloalcohol dehalogenase(HheC) Epichlorohydrin epoxide hydrolase (echA) into model plant- Nicotiana tabacum and transform 1,2,3-TCP into glycerol.