典型环境污染物修复基因工程菌的构建及应用

随着人类活动的增加,对有机物和重金属的应用越来越广泛,同时造成的环境污染程度越来越严重.综述了石油、农药、表面活性剂及重金属类污染物治理中基因工程菌的构建及应用的研究进展,指出利用基因工程菌解决环境中的石油、农药、表面活性剂及重金属的污染问题已成为环境污染修复领域的研究热点,并提出基因工程菌的构建及应用过程中的难点及发展趋势.     

A comprehensive overview of elements in bioremediation

Sustainable development requires the development and promotion of environmental management and a constant search for green technologies to treat a wide range of aquatic and terrestrial habitats contaminated by increasing anthropogenic activities. Bioremediation is an increasingly popular alternative to conventional methods for treating waste compounds and media with the possibility to degrade contaminants using natural microbial activity mediated by different consortia of microbial strains. Many studies about bioremediation have been reported and the scientific literature has revealed the progressive emergence of various bioremediation techniques. In this review, we discuss the various in situ and ex situ bioremediation techniques and elaborate on the anaerobic digestion technology, phytoremediation, hyperaccumulation, composting and biosorption for their effectiveness in the biotreatment, stabilization and eventually overall remediation of contaminated strata and environments. The review ends with a note on the recent advances genetic engineering and nanotechnology have had in improving bioremediation. Case studies have also been extensively revisited to support the discussions on biosorption of heavy metals, gene probes used in molecular diagnostics, bioremediation studies of contaminants in vadose soils, bioremediation of oil contaminated soils, bioremediation of contaminants from mining sites, air sparging, slurry phase bioremediation, phytoremediation studies for pollutants and heavy metal hyperaccumulators, and vermicomposting.     

Expression analysis in response to low temperature stress in blood oranges: implication of the flavonoid biosynthetic pathway

This minireview explores the environmental bioremediation mediated by genetically engineered (GE) bacteria and it also highlights the limitations and challenges associated with the release of engineered bacteria in field conditions. Application of GE bacteria based remediation of various heavy metal pollutants is in the forefront due to eco-friendly and lesser health hazards compared to physico-chemical based strategies, which are less eco-friendly and hazardous to human health. A combination of microbiological and ecological knowledge, biochemical mechanisms and field engineering designs would be an essential element for successful in situ bioremediation of heavy metal contaminated sites using engineered bacteria. Critical research questions pertaining to the development and implementation of GE bacteria for enhanced bioremediation have been identified and poised for possible future research. Genetic engineering of indigenous microflora, well adapted to local environmental conditions, may offer more efficient bioremediation of contaminated sites and making the bioremediation more viable and eco-friendly technology. However, many challenges are to be addressed concerning the release of genetically engineered bacteria in field conditions. There are possible risks associated with the use of GE bacteria in field condition, with particular emphasis on ways in which molecular genetics could contribute to the risk mitigation. Both environmental as well as public health concerns need to be addressed by the molecular biologists. Although bioremediation of heavy metals by using the genetically engineered bacteria has been extensively reviewed in the past also, but the bio-safety assessment and factors of genetic pollution have been never the less ignored.     

合成生物学改造微生物及生物被膜用于重金属污染检测与修复

随着工业化进程不断加快,重金属污染日益加剧,尤其是水体的重金属污染,已严重威胁人类健康,迫切需要进行有效的污染修复.相比传统物理和化学修复,生物修复具有绿色环保和可持续性的特点.因为微生物生长繁殖迅速、生物被膜具有动态可调节和环境适应性好等特点,使其能更好耐受胁迫环境,在环境修复中有重要作用.合成生物学改造微生物及生物...     

根系分泌物及其在植物修复中的作用

 近年来环境污染日益严重,污染物在土壤植物中的行为引起了人们的高度关注。利用植物去除土壤水体等介质中污染物的植物修复是近10年来兴起的一项安全、廉价的技术,已成为污染生态学和环境生态学的研究热点,它通过植物吸收、根滤、稳定、挥发等方式清除环境中的重金属和有机污染物。国内外有关植物修复的研究报道和概述很多, 但对植物根系分泌物在植物修复中所起的作用及其机理少有述评。 本文从根系分泌物对土壤重金属和土壤有机污染物的去除作用出发,对根系分泌物的种类、数量及其在去除环境污染物中的作用机理和功能地位进行了总结,并借助研究事例对影响植物根系分泌的内外因子,如植物种类、营养胁迫、重金属胁迫、根际环境的理化性质、土壤微生物及其它环境因子进行了讨论。概言之,根系分泌物在修复污染土壤中的重金属途径是多种多样的,主要是通过调节根际pH值、与重金属形成螯合物、络合反应、沉淀、提高土壤微生物数量和活性来改变重金属在根际中的存在形态以及提高重金属的生物有效性,从而减轻它对环境的危害。在清除有机污染物时,根系分泌物中的酶可以对有机污染物进行直接降解,根系分泌物影响下的微生物也可以对有机污染物进行间接降解,且被认为是主要的降解途径。根系分泌物在植物修复过程中确实起着某些重要作用,今后应将这方面的研究重点放在某些特异性根系分泌物植物,尤其是某些重金属超富集植物资源的寻找、筛选上,通过室内实验和野外研究确定其根系分泌物对清除重金属和有机污染物的效率,证实超富集植物根系分泌物的特异性与污染物超富集的内在联系,找到污染土壤生态恢复和治理的有效方法并加以推广应用,如针对性地在被污染地大面积种植此类具特异性根分泌物植物,并辅以营林措施如修剪等,加快生物修复进程,提高修复效率。植物根系分泌物在植物修复过程中所具有的重要生态意义和可能应用前景,为污染生态学和化学生态学之间的联合研究开拓了全新的领域,今后将取得新的突破和重要进展。     

Bioremediation 3.0: Engineering pollutant-removing bacteria in the times of systemic biology

Elimination or mitigation of the toxic effects of chemical waste released to the environment by industrial and urban activities relies largely on the catalytic activities of microorganisms—specifically bacteria. Given their capacity to evolve rapidly, they have the biochemical power to tackle a large number of molecules mobilized from their geological repositories through human action (e.g., hydrocarbons, heavy metals) or generated through chemical synthesis (e.g., xenobiotic compounds). Whereas naturally occurring microbes already have considerable ability to remove many environmental pollutants with no external intervention, the onset of genetic engineering in the 1980s allowed the possibility of rational design of bacteria to catabolize specific compounds, which could eventually be released into the environment as bioremediation agents. The complexity of this endeavour and the lack of fundamental knowledge nonetheless led to the virtual abandonment of such a recombinant DNA-based bioremediation only a decade later. In a twist of events, the last few years have witnessed the emergence of new systemic fields (including systems and synthetic biology, and metabolic engineering) that allow revisiting the same environmental pollution challenges through fresh and far more powerful approaches. The focus on contaminated sites and chemicals has been broadened by the phenomenal problems of anthropogenic emissions of greenhouse gases and the accumulation of plastic waste on a global scale. In this article, we analyze how contemporary systemic biology is helping to take the design of bioremediation agents back to the core of environmental biotechnology. We inspect a number of recent strategies for catabolic pathway construction and optimization and we bring them together by proposing an engineering workflow.     

Plant growth promoting rhizobacteria and endophytes accelerate phytoremediation of metalliferous soils

Technogenic activities (industrial—plastic, textiles, microelectronics, wood preservatives; mining—mine refuse, tailings, smelting; agrochemicals—chemical fertilizers, farm yard manure, pesticides; aerosols—pyrometallurgical and automobile exhausts; biosolids—sewage sludge, domestic waste; fly ash—coal combustion products) are the primary sources of heavy metal contamination and pollution in the environment in addition to geogenic sources. During the last two decades, bioremediation has emerged as a potential tool to clean up the metal-contaminated/polluted environment. Exclusively derived processes by plants alone (phytoremediation) are time-consuming. Further, high levels of pollutants pose toxicity to the remediating plants. This situation could be ameliorated and accelerated by exploring the partnership of plant-microbe, which would improve the plant growth by facilitating the sequestration of toxic heavy metals. Plants can bioconcentrate (phytoextraction) as well as bioimmobilize or inactivate (phytostabilization) toxic heavy metals through in situ rhizospheric processes. The mobility and bioavailability of heavy metal in the soil, particularly at the rhizosphere where root uptake or exclusion takes place, are critical factors that affect phytoextraction and phytostabilization. Developing new methods for either enhancing (phytoextraction) or reducing the bioavailability of metal contaminants in the rhizosphere (phytostabilization) as well as improving plant establishment, growth, and health could significantly speed up the process of bioremediation techniques. In this review, we have highlighted the role of plant growth promoting rhizo- and/or endophytic bacteria in accelerating phytoremediation derived benefits in extensive tables and elaborate schematic sketches.     

Isolation and characterization of heavy metal tolerant Gram-positive bacteria with bioremedial properties from municipal waste rich soil of Kestopur canal (Kolkata), West Bengal,India

The present study was conducted to determine the culturable bacterial profile from Kestopur canal (Kolkata, India) and analyze their heavy metal tolerance. In addition to daily sewage including solid and soluble wastes, a considerable load of toxic metals are released into this water body from industries, tanneries and agriculture, household as well as health sectors. Screening out microbes from such an environment was done keeping in mind their multifunctional application especially for bioremediation. Heavy metals are major environmental pollutants when present in high concentration in soil and show potential toxic effects on growth and development in plants and animals. Some edible herbs growing in the canal vicinity, and consumed by people, were found to harbour these heavy metals at sub-toxic levels. The bioconcentration factor of these plants being <1 indicates that they probably only absorb but not accumulate heavy metals. All the thirteen Grampositive bacteria isolated from these plants rhizosphere were found to tolerate high concentration of heavy metals like Co, Ni, Pb, Cr, Fe. Phylogenetic analysis of their 16S rDNA genes revealed that they belonged to one main taxonomic group — the Firmicutes. Seven of them were found to be novel with 92–95% sequence homology with known bacterial strains. Further microbiological analyses show that the alkaliphilic Bacillus weihenstephanensis strain IA1 and Exiguobacterium aestuarii strain CE1, with selective antibiotic sensitivity along with high Ni²⁺ and Cr⁶⁺ removal capabilities, respectively, can be prospective candidates for bioremediation.     

Removal of Heavy Metals from the Environment by Biosorption

The pollution of the environment with toxic metals is a result of many human activities, such as mining and metallurgy, and the effects of these metals on the ecosystems are of large economic and public‐healthsignificance. This paper presents the features and advantages of the unconventional removal method of heavy metals – biosorption – as a part of bioremediation. Bioremediation consists of a group of applications, which involvethe detoxification of hazardous substances instead of transferring them from one medium to another, by means of microbes and plants. This process is characterized as less disruptive and can be often carried out on site, eliminating the need to transport the toxic materials to treatment sites. The biosorption (sorption of metallic ions from solutions by live or dried biomass) offers an alternative to the remediation of industrial effluents as well as the recovery of metals contained in other media. Biosorbents are prepared from naturally abundant and/or waste biomass. Due to the high uptake capacity and very cost‐effective source of the raw material, biosorption is a progression towards a perspective method. The mechanism by which microorganisms take up metals is relatively unclear, but it has been demonstrated that both living and non‐living biomass may be utilized in biosorptive processes, as they often exhibit a marked tolerance towards metals and other adverse conditions. One of their major advantages is the treatment of large volumes of effluents with low concentrations of pollutants. Models developed were presented to determine both the number of adsorption sites required to bind each metal ion and the rate of adsorption, using a batch reactor mass balance and the Langmuir theory of adsorption to surfaces or continuous dynamic systems. Two main categories of bioreactors used in bioremediation – suspended growth and fixed film bioreactors – are discussed. Reactors with varying configurations to meet the different requirements for biosorption are analyzed considering two major groups of reactors – batch reactors and continuous reactors. Biosorption is treated as an emerging technology effective in removing even very low levels of heavy metal.     

Phytoremediation--a novel and promising approach for environmental clean-up

Phytoremediation is an eco friendly approach for remediation of contaminated soil and water using plants. Phytoremediation is comprised of two components, one by the root colonizing microbes and the other by plants themselves, which degrade the toxic compounds to further non-toxic metabolites. Various compounds, viz. organic compounds, xenobiotics, pesticides and heavy metals, are among the contaminants that can be effectively remediated by plants. Plant cell cultures, hairy roots and algae have been studied for their ability to degrade a number of contaminants. They exhibit various enzymatic activities for degradation of xenobiotics, viz. dehalogenation, denitrification leading to breakdown of complex compounds to simple and non-toxic products. Plants and algae also have the ability to hyper accumulate various heavy metals by the action of phytochelatins and metallothioneins forming complexes with heavy metals and translocate them into vacuoles. Molecular cloning and expression of heavy metal accumulator genes and xenobiotic degrading enzyme coding genes resulted in enhanced remediation rates, which will be helpful in making the process for large-scale application to remediate vast areas of contaminated soils. A few companies worldwide are also working on this aspect of bioremediation, mainly by transgenic plants to replace expensive physical or chemical remediation techniques. Selection and testing multiple hyperaccumulator plants, protein engineering ofphytochelatin and membrane transporter genes and their expression would enhance the rate of phytoremediation, making this process a successful one for bioremediation of environmental contamination. Recent years have seen major investments in the R&D, which have also resulted in competition of filing patents by several companies for economic gains. The details of science & technology related to phytoremediation have been discussed with a focus on future trends and prospects of global relevance.     

Biological treatments for contaminated soils: hydrocarbon contamination. Fungal applications in bioremediation treatment

Bioremediation is a spontaneous or controlled process in which biological, mainly microbiological, methods are used to degrade or transform contaminants to non or less toxic products, reducing the environmental pollution. The most important parameters to define a contaminated site are: biodegradability, contaminant distribution, lixiviation grade, chemical reactivity of the contaminants, soil type and properties, oxygen availability and occurrence of inhibitory substances. Biological treatments of organic contaminations are based on the degradative abilities of the microorganisms. Therefore the knowledge on the physiology and ecology of the biological species or consortia involved as well as the characteristics of the polluted sites are decisive factors to select an adequate biorremediation protocol. Basidiomycetes which cause white rot decay of wood are able to degrade lignin and a variety of environmentally persistent pollutants. Thus, white rot fungi and their enzymes are thought to be useful not only in some industrial process like biopulping and biobleaching but also in bioremediation. This paper provides a review of different aspects of bioremediation technologies and recent advances on ligninolytic metabolism research.     

Histochemical properties and trace element concentrations in <Emphasis Type="Italic">Parietaria L.</Emphasis>from urban sites (Palermo,Italy)

We examined leaf and pollen histochemical properties in Parietaria L. and measured trace element concentrations in urban sites in Palermo, Sicily, affected by different stress valence levels. Urban pollution affects histochemistry macro-, micro-, and toxic element concentrations in Parietaria. We therefore decided to determine whether this plant can be used as a simple biomonitor or bioaccumulator of pollution contaminants. Samples were collected at various sites in different periods. An increase in phenolic compound levels was a result of the impact of pollution. Histochemical characteristics and trace element concentrations varied according to the period, but not to the place, of sampling. Pollutant concentrations were the highest in the month of February 2002. Trace metal concentrations were the lowest in July. Our investigations have confirmed that Parietaria L. can be used to study the effects of trace elements in the atmosphere in urban sites; in fact, they are veritable accumulators, given that elevated levels of pollutants can be found on their structures with no visible morphological effects. There are differences in accumulation capacity according to the trace elements studied. Pollution alters the content of phenolic compounds and chemical elements in Parietaria L.     

Phytoremediation—A Novel and Promising Approach for Environmental Clean-up

ABSTRACT

Phytoremediation is an eco friendly approach for remediation of contaminated soil and water using plants. Phytoremediation is comprised of two components, one by the root colonizing microbes and the other by plants themselves, which degrade the toxic compounds to further non-toxic metabolites. Various compounds, viz. organic compounds, xenobiotics, pesticides and heavy metals, are among the contaminants that can be effectively remediated by plants. Plant cell cultures, hairy roots and algae have been studied for their ability to degrade a number of contaminants. They exhibit various enzymatic activities for degradation of xenobiotics, viz. dehalogenation, denitrification leading to breakdown of complex compounds to simple and non-toxic products. Plants and algae also have the ability to hyper accumulate various heavy metals by the action of phytochelatins and metallothioneins forming complexes with heavy metals and translocate them into vacuoles. Molecular cloning and expression of heavy metal accumulator genes and xenobiotic degrading enzyme coding genes resulted in enhanced remediation rates, which will be helpful in making the process for large-scale application to remediate vast areas of contaminated soils. A few companies worldwide are also working on this aspect of bioremediation, mainly by transgenic plants to replace expensive physical or chemical remediation techniques. Selection and testing multiple hyperaccumulator plants, protein engineering of phytochelatin and membrane transporter genes and their expression would enhance the rate of phytoremediation, making this process a successful one for bioremediation of environmental contamination. Recent years have seen major investments in the R&D, which have also resulted in competition of filing patents by several companies for economic gains. The details of science & technology related to phytoremediation have been discussed with a focus on future trends and prospects of global relevance.     

重金属污染土壤植物修复中的微生物功能研究进展

综述了国内外在重金属污染土壤植物-微生物联合修复领域的研究报道,总结了近5年的研究实例。植物-微生物联合修复体系具有生物固定与生物去除土壤重金属的两种功能,根际微生物可以菌根、内生菌等方式与根系形成联合体,通过增强植物抗性和优化根际环境,促进根系发展,增强植物吸收和向上转运重金属的能力。建立植物-微生物联合修复体系,可充分发挥植物与微生物作用功能的优势,提高污染土壤的修复效率。增强植物修复体系中微生物功能的重点是深入研究根际微生物、根系和介质载体三者之间复合功能,结合污染土壤类型与植物群落配置的特点筛选扩繁高效菌种与菌群。     

Effect of heavy metal pollution on mycorrhizal colonization and function: physiological, ecological and applied aspects

 High concentrations of heavy metals in soil have an adverse effect on micro-organisms and microbial processes. Among soil microorganisms, mycorrhizal fungi are the only ones providing a direct link between soil and roots, and can therefore be of great importance in heavy metal availability and toxicity to plants. This review discusses various aspects of the interactions between heavy metals and mycorrhizal fungi, including the effects of heavy metals on the occurrence of mycorrhizal fungi, heavy metal tolerance in these micro-organisms, and their effect on metal uptake and transfer to plants. Mechanisms involved in metal tolerance, uptake and accumulation by mycorrhizal hyphae and by endo- or ectomycorrhizae are covered. The possible use of mycorrhizal fungi as bioremediation agents in polluted soils or as bioindicators of pollution is also discussed. Accepted: 23 June 1997     

AM真菌对重金属污染土壤生物修复的应用与机理

土壤重金属污染威胁人类健康和整个生态系统,而高效、低耗、安全的生物修复技术显示出了极大的应用潜力,特别是利用植物-微生物共生体增强生物修复效应的应用.丛枝菌根(Arbuscular Mycorrhizae,AM)真菌是一类广泛分布于土壤生态系统中的有益微生物,能与90％以上的陆生高等植物形成共生体.研究发现,AM真菌能够增强宿主植物对土壤中重金属胁迫的耐受性.当前,利用AM真菌开展重金属污染土壤的生物修复已经引起环境学家和生态学家的广泛关注.基于此,围绕AM真菌在重金属污染土壤生物修复作用中的最新研究进展,从物理性防御体系的形成、对植物生理代谢的调控、生化拮抗物质的产生、基因表达的调控等角度探究AM真菌在重金属污染土壤生物修复中的作用机理,以期为利用AM真菌开展重金属污染的生物修复提供理论依据,并对本领域未来的发展和应用前景进行了展望.     

Applications of biofilms in bioremediation and biotransformation of persistent organic pollutants, pharmaceuticals/personal care products, and heavy metals

In this review, the strategies being employed to exploit the inherent durability of biofilms and the diverse nutrient cycling of the microbiome for bioremediation are explored. Focus will be given to halogenated compounds, hydrocarbons, pharmaceuticals, and personal care products as well as some heavy metals and toxic minerals, as these groups represent the majority of priority pollutants. For decades, industrial processes have been creating waste all around the world, resulting in contaminated sediments and subsequent, far-reaching dispersal into aquatic environments. As persistent pollutants have accumulated and are still being created and disposed, the incentive to find suitable and more efficient solutions to effectively detoxify the environment is even greater. Indigenous bacterial communities are capable of metabolizing persistent organic pollutants and oxidizing heavy metal contaminants. However, their low abundance and activity in the environment, difficulties accessing the contaminant or nutrient limitations in the environment all prevent the processes from occurring as quickly as desired and thus reaching the proposed clean-up goals. Biofilm communities provide among other things a beneficial structure, possibility for nutrient, and genetic exchange to participating microorganisms as well as protection from the surrounding environment concerning for instance predation and chemical and shear stresses. Biofilms can also be utilized in other ways as biomarkers for monitoring of stream water quality from for instance mine drainage. The durability and structure of biofilms together with the diverse array of structural and metabolic characteristics make these communities attractive actors in biofilm-mediated remediation solutions and ecosystem monitoring.     

金属结合蛋白(肽)与环境重金属生物修复

重金属污染是全球关注的重要环境问题。针对重金属的生物修复技术 ,因其特有的优势 ,越来越受到重视 ,其中一个重要的研究领域是利用金属离子和金属结合蛋白或结合肽之间存在的强亲和能力特性进行的生物修复研究。就金属结合蛋白 (肽 )的种类、结构特点、以及金属结合的作用机理进行了总结 ,同时综述了展示或表达有不同金属结合蛋白或结合肽的微生物和植物对重金属污染进行生物修复的最新研究进展 ,对基于金属结合蛋白 (肽 )的环境重金属生物修复的进一步研究 (如肽库的构建和筛选 ,金属与蛋白 (肽 )的相互作用 )进行了讨论。     

Assessing the applicability of phytoremediation of soils with mixed organic and heavy metal contaminants

Soil pollution is a major environmental problem and many contaminated sites are tainted with a mixture of organic and heavy metal contaminants. Compared to other remedial strategies, phytoremediation is a low cost, environmentally-friendly, sustainable means of remediating the contamination. This review first provides an overview of phytoremediation studies where the soil is contaminated with just one type of pollutant (heavy metals or organics) and then critically evaluates the applicability of phytotechnologies for the remediation of contaminated sites where the soil is polluted by a mixture of organic and heavy metal contaminants. In most of the earlier research studies, mixed contamination was held to be detrimental to plant growth, yet there were instances where plant growth was more successful in soil with mixed contamination than in the soil with only individual contaminants. New effective phytoremediation strategies can be designed for remediation of co-contaminated sites using: (a) plants species especially adapted to grow in the contaminated site (hyperacumulators, local plants, transgenic plants); (b) endophytic bacteria to enhance the degradation in the rizhosphere; (c) soil amendments to increase the contaminants bioavailability [chelating agents and (bio)surfactants]; (d) soil fertilization to enhance the plant growth and microbial activity in the soil; and (e) coupling phytoremediation with other remediation technologies such as electrokinetic remediation or enhanced biodegradation in the rhizosphere.