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    PSB464 - PLANT ENZYMES INVOLVED IN PHYTOREMEDIATION; ROLE OF MICROORGANISMS IN BIODEGRADATION-1

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    PSB464 - Plant Enzymes Involved in Phytoremediation Role of Microorganisms in Biodegradation-1

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    | PSB 4 HYTOREMEDIATION AND EROSION CONTROL (BY OKUH-IKEME, P.C) TOPIC 2: PLANT ENZYMES INVOLVED IN PHYTOREMEDIATION INTRODUCTION: Environmental pollution is growing more and more due to the indiscriminate and frequently deliberate release of hazardous, harmful substances, Research efforts have been devoted to develop new, low-cost, low- technology, eco-friendly treatments capable of reducing and even eliminating pollution in the atmosphere, the hydrosphere and soil environments. Among biological agents, enzymes have a great potentiality to effectively transform and detoxify polluting substances because they have been recognized to be able to transform pollutants at a detectable rate and are potentially suitable to restore polluted environments. A large number of enzymes from bacteria, fungi, and plants have been reported to be involved in the biodegradation of toxic organic pollutants, Enzymes have several beneficial characteristics: They are the main effectors of all the transformations occurring in the biota. They are catalysts with either narrow or broad specificity and, therefore, can be applied to a large range of different compounds in mixture. -They may produce extensive transformations of structural and toxicological properties of contaminants and even their complete conversion into innocuous inorganic end products. ~They may perform processes for which no efficient chemical transformations have been devised. The most representative enzymatic classes in the remediation of polluted environments are: © Hydrolases: examples- phosphotriesterases, amudases, proteases, carbohydrases (cellulases and amylases), depolymerase * Dehalogenases © Transferases * Oxidoreductases: Mono- ‘or di-oxygenases, reductases, dehalogenases, cytochrome P450 monoxygenases, phenoloxidases (laccases, tyrosinases) and peroxidases (lignin and manganese peroxidases) are the main classes of oxidoreductases. Their main producers are bacteria, fungi, mainly white-rot fungi, plants and microbe-plant associations. For many of these enzymes the transformation of different xenobiotic substances has been tested mainly under laboratory conditions. OXIDOREDUCTASE These enzymes participate in the oxidation reaction which reduce substrates by transferring oxygen from molecular oxygen by utilizing FADINADH/NADPH as cofactors. Oxidoreductases follow the reactions A"+ B = A+B where A is the oxidant and B, the reductant. The toxicity of these pollutants can be remediated by various bacteria as well as higher plants and is driven by enzyme oxidoreductases. By transferring the electrons from the substrate to the compound, it helps to break chemical bonds and finally the contamination is oxidized to harmless organic compounds. Lignin decomposition in a soil environment with humification of phenolic substances occurs through this enzyme. In addition, it can also detoxify xenobiotic compounds through the processes of binding to humic substances and polymerization. Most plant families such as Fabaceae and Solanaceae release oxidoreductases, which are involved in the detoxification process in soil. Various microbial enzymes (namely, oxygenases, reductases, dehalogenases, dehydrogenases, and hydroxylascs) play an important role in remediation of environmental pollutants which have been distributed in the biosphere due to anthropogenic activities. Among these enzymes, oxygenases are important enzymes because they are primarily involved in the initial process of degradation and catalyze and degrade the aromatic compounds. They catalyze by the addition of oxygen atoms into the substrates, Two major classes of oxygenases have been identified: monooxygenases (addition of one oxygen molecule) and dioxygenases (addition of two oxygen molecules). -Microbial monooxygenases: In substrate, there is incorporation of a single oxygen molecule with the help of enzyme monooxygenases. On the basis of the cofactors, it is categorized into two subclasses: Flavin-dependent monooxygenases and (2) P450 monooxygenases (e.g., CYP102 from Bacillus megaterium BM3). The prosthetic group of the first subclass is flavin, which is activated by the coenzymes NADP or NADPH, while the second subclass contains heme. Monooxygenases are cofactor-dependent biocatalysts involved in bioremediation. Some enzymes are cofactor-independent and perform their activity with the molecular oxygen only. A number of processes, such as desulfurization, denitrification, nitrification, ammonification, dehalogenation, transformation, hydroxylation, and aromatic and aliphatic biodegradation are regulated by enzyme monooxygenases, for example, ActVAorf6 monooxygenase, TemH, etc. Methane monooxygenase enzyme is the best characterized as playing a role in the degradation of hydrocarbons. -Microbial dioxygenases: These are multicomponent enzyme systems which link molecular oxygen to the substrate. They degrade the aromatic compounds that cause serious environmental damage. On the basis of the mode of action of the enzyme, it can categorized into two subclasses: hydroxylation and cleavage dioxygenases. The hydroxylation enzyme catalyzes the addition of two oxygen atoms into the substrate, whereas the cleavage enzyme catalyzes an aromatic ring typically carrying two or more hydroxyl groups. The cleavage dioxygenases are further classified into two groups, intradiol (ortho cleavage and utilize FellI) and extradiol (Meta cleavage and utilize Fell and Mnll). These enzymes are involved in degrading aromatic molecules in the environment. They are soil bacteria which are involved in the transformation process by changing aromatic precursors into aliphatic products MICROBIAL LACCASES These are the oxidases of blue multicopper which act as biosensors which catalyze substrate oxidation. Most of these microorganisms have the potential to synthesize intracellular laccases and extracellular laccases involved in the oxidation of aminophenols, lignins, ortho and paradiphenols, polyamines, polyphenols, aryl diamines, etc. Laccases decolorize and detoxify industrial effluents and help in wastewater treatment. They act on highly environmental contaminant, phenolic and nonphenolic organic compounds and are valuable for the textile, pulp, and paper industries. In addition, they are directly involved in the degradation of xenobiotics and bioremediation. In addition, thermophilic laccase enzyme, which was isolated from desert plant species such as Cereus spp. and Opuntia spp., has shown thermophilic properties and is utilized in the pulping industry. MICROBIAL PEROXIDASES Peroxidases are widely distributed in nature. These enzymes are produced by a variety of sources including plants, animals, and microbes. They are produced from different sources including fungi, bacteria, yeast, and Cyanobacteria. These microbial enzymes are involved in the degradation of environmental contaminant, feedstock of animal, and raw materials for the chemical, agricultural, and paper industries, textile dye degradation, paper/pulp industry for lignin degradation, dye decolorization, sewage treatment, and also as biosensors. In plants, they are involved in lignin synthesis, cell wall formation, metabolism of auxin, cell elongation, and defense mechanism. They are also subdivided into heme and nonheme proteins. Hemeperoxidases are divided into two major groups which include prokaryotes as well as eukaryotes. Peroxidases are again subdivided into three classes based on the comparison. -Class I: This is an intracellular enzyme which has ascorbate peroxidase, yeast cytochrome ¢ peroxidase, as well as catalase peroxidases. -Class II: This is involved in the lignin degradation of wood which consists of fungal peroxidases, namely, manganese peroxidase (Mnp) and lignin peroxidase. “Class III: This category of enzyme is involved in the formation of cellwall and lignifications and contains plant peroxidases, namely, horse radish, barley, or soybean. icrobial lignin: during secondary metabolism these heme proteins are secreted by white-rot fungus and legrade the cell wall of plants containing lignin. -Microbial Mnps: these are extracellular heme enzymes. In a multistep reaction they catalyze the oxidation of Mn21 to Mn31 oxidants. A reduced form of Mn enhanced the production of Mnps and the oxidant acts as a precursor for phenolic compound oxidation. -Microbial versatile peroxidases: these helps to oxidize methoxybenzenes, Mn21, phenolic compounds, and dimers of lignin. Therefore they are required for the removal of environmental contaminants. HYDROLYTI ZYMES FOR BIOREMEDIATION Besides the abovementioned enzymes, some hydrolytic enzymes are also important in the bioremediation of soil pollutants. Water is contaminated by industrial waste, petroleum waste, and degraded by some hydrolytic enzymes such as proteases, lipases, pullulanases, amylases, and xylanases. These enzymes have a diverse role in different areas including in different industries and biochemical sciences. LIPASES Lipase is present in plants and animals as well as microorganisms, and plays an important role in lipid degradation. It acts on the soil organic contaminants and helps in the reduction of pollutants from the contaminated soil; the reactions include esterification, aminolysis, hydrolysis, etc. It is an indicator to degrade hydrocarbon pollutants from soil. Lipase has various applications in the cosmetic, food, paper, pulp, food and chemical industries, etc. but the cost of production is restrictive, AMYLASES Alpha amylases are an extracellular enzyme which breaks the d-1,4-glucosidic linkage in starch molecules and produces oligosaccharides. -Amylase also breaks the second a-,4-glycosidic bond of maltose and is synthesized in plants as well as bacteria. Amylases are significant enzymes for their specific use in the industrial starch conversion process. These enzymes act particularly on disaccharides (suctose) and polysaccharides (starch) and are categorized in the glycoside hydrolases group. Application of these enzymes has been established in the starch liquefaction, paper, food, sugar, and pharmaceutical industries, PROTEASES These are a group of hydrolytic enzymes which cleave peptide bonds that occur within the primary structure of polypeptides and other proteins as well. They are well known industrial enzymes, with a key role in different industries, for example, pharmaceutical, textile, feed food, and can be extracted from plants, animals, and microorganisms. Protease provides potential applications for the management of waste from some processing industries and household activities. In addition, itis also helpful in poultry processing industries. PULLULANASE a Dextrin 6-glucanohydrolases or pullulanases is synthesized from several microorganisms such as Klebsiella spp., Bacillus spp., and Geobacillus stearothermophilus. Due to its specific enzymatic action on pullulan, particularly in the specific linkages (a-1,6 linkages), itis very popular. It is very important for its action as a bioprocessor of starch. Pullulanase enzyme has a wide role in genetic engineering and some industrial uses. It has five groups: 1. pullulanase type J; 2. hydrolase type III; 3. neopullulanase; 4, amylopullulanase; and 5. isopullulanase TOPIC 3: THE ROLE OF MICROORGANI: BIODEGRADATIO! DUCTION: Microorganisms are widely distributed on the biosphere because their metabolic ability is very impressive and they can easily grow in a wide range of environmental conditions. The nutritional versatility of microorganisms can also be exploited for biodegradation of pollutants. This kind of process is termed as bioremediation. It is based on the ability of certain microorganisms to convert, modify and utilize toxic pollutants in order obtain energy and produce biomass. Instead of simply collecting the pollutant and storing it, bioremediation is @ microbiological well organized procedural activity which is applied to break down or transform contaminants to less toxic or non-toxic elemental and compound forms, The process of bioremediation mainly depends on microorganisms which enzymatically attack the pollutants and convert them to innocuous products. As its bioremediation can be effective only where environmental conditions permit microbial growth and activity, low microbial growth and application often involves the manipulation of environmental parameters to all degradation to proceed at a faster rate. Biodegradation is defined as the biologically catalyzed reduction in complexity of chemical compounds. It is the process by which organic substances are broken down into smaller compounds by living microbial ". The microbial organisms organisms. When biodegradation is complete, the process is called "mineralization transform the substance through metabolic or enzymatic processes. It is based on two processes: growth and cometabolism. In growth, an organic pollutant is used as sole source of carbon and energy. This process results ina complete degradation (mineralization) of organic pollutants. Cometabolism is defined as the metabolism of an organic compound in the presence of a growth substrate that is used as the primary carbon and energy source, Several microorganisms, including fungi, bacteria and yeasts are involved in biodegradation process. Biodegradable matter is generally organic material such as plant and animal matter and other substances originating from living organisms, or artificial materials that are similar enough to plant and animal matter to be put to use by microorganisms. Some microorganisms have the astonishing, naturally occurring, microbial catabolic diversity to degrade, transform or accumulate a huge range of compounds including hydrocarbons (e.g. oil), polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), radionuclides and metals, Biodegradation is nature's way of recycling wastes, or breaking down organic matter into nutrients that can be used and reused by other organisms. In the microbiological sense, "biodegradation" means that the decaying of all organic materials is carried out by a huge assortment of life forms comprising mainly bacteria, yeast and fungi, and possibly other organisms. Bioremediation and biotransformation methods endeavour to hamess the astonishing, naturally occurring, microbial catabolic diversity to degrade, transform or accumulate a huge range of compounds including hydrocarbons (e.g, oil), polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), radionuclides and metals. Bioremediation process can be divided into three phases or levels. First, through natural attenuation, contaminants are reduced by native microorganisms without any human augmentation. Second, biostimulation is employed where nutrients and oxygen are applied to the systems to improve their effectiveness and to accelerate biodegradation. Finally, during bioaugmentation, microorganisms are added to the systems. These supplemental organisms should be more efficient than native flora to degrade the target contaminant. A feasible remedial technology requires microorganisms being capable of quick adaptation and efficient uses of pollutants of interest in a particular case in a reasonable period of time. Many factors influence the ability of microorganisms to use pollutants as substrates: the genetic potential and certain environmental factors such as temperature, pH, and available nitrogen and phosphorus sources. Applications of genetically engineered microorganisms (GEM) in bioremediation have received a great deal of attention. These GEM have higher degradative capacity and have been demonstrated successfully for the degradation of various pollutants under defined conditions. However, ecological and environmental concems and regulatory constraints are major obstacles for testing GEM in the field. polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), pesticides and dyes are some of these types of compounds. Some other synthetic chemicals like radionuclides and metals are extremely resistant to biodegradation by native flora compared with the naturally occurring organic compounds that are readily degraded upon introduction into the environment. -Hydrocarbons: These are organic compounds whose structures consist of hydrogen and carbon. Hydrocarbons can be seen as linear linked, branched or cyclic molecules. They are observed as aromatic or aliphatic hydrocarbons. The first one has benzene (CsHe) in its structure, while the aliphatic one is seen in three forms: alkanes, alkenes and alkynes. -Polycyclic aromatic hydrocarbons (PAHs): These are important pollutants class of hydrophobic organic contaminants (HOCs) widely found in air, soil and sediments. The major source of PAH pollution is industrial production. They have been studied with increasing interest for more than twenty years because of more findings about their toxicity, environmental persistence and prevalence. PAHs can be absorbed into organic-rich soils and sediments, accumulate in fish and other aquatic organisms, and may be transferred to humans through seafood consumption. The biodegradation of PAHs can be considered on one hand to be part of the normal processes of the carbon cycle, and on the other as the removal of man-made pollutants from the environment. The use of microorganisms for bioremediation of PAH-contaminated environments seems to be an attractive technology for restoration of polluted sites. -Polychlorinated biphenyls (PCBs): These are mixtures of synthetic organic chemicals. Due to their non- flammability, chemical stability, high boiling point, and electrical insulating properties, PCBs were used in hundreds of industrial and commercial applications including electrical, heat transfer, and hydraulic equipment, as plasticizers in paints, plastics, and rubber products; in pigments, dyes, and carbonless copy paper; and many other industrial applications. Consequently, PCBs are toxic compounds that could act as endocrine disrupters and cause cancer. Therefore, environmental pollution with PCBs is of increasing concern. -Pesticides: These are substances or mixture of substances intended for preventing, destroying, repelling or mitigating any pest. Pesticides which are rapidly degraded are called non-persistent while those which resist degradation are termed persistent, The most common type of degradation is carried out in the soil by microorganisms, especially fungi and bacteria that use pesticides as food source. “Dyes: These are widely used in the textile, rubber product, paper, printing, colour photography, pharmaceuticals, cosmetics and many other industries. Azo dyes, which are aromatic compounds with one or more (-N=N-) groups, are the most important and largest class of synthetic dyes used in commercial applications, These dyes are poorly biodegradable because of their structures and treatment of wastewater containing dyes usually involves physical and / or chemical methods such as adsorption, coagulation- flocculation, oxidation, filtration and electrochemical methods. The success of a biological process for colour removal from a given effluent depends in part on the utilization of microorganisms that effectively decolorize synthetic dyes of different chemical structures. “Radionuclides: A radionuclide is an atom with an unstable nucleus, characterized by excess eneruy available to be imparted either to a newly created radiation particle within the nucleus or via internal conversion. During this process, the radionuclide is said to undergo radioactive decay, resulting in the emission of gamma ray(s) and/or subatomic particles such as alpha or beta particles. -Heavy metals: Unlike organic contaminants, the metals cannot be destroyed, but must either be converted to a stable form or removed. Bioremediation of metals is achieved through biotransformation. Mechanisms by Which microorganisms act on heavy metals include bioabsorption (metal absorption to cell surface by physicochemical mechanisms), bioleaching (heavy metal mobilization through the excretion of organic acids or methylation reactions), biomineralization (heavy metal immobilization through the formation of insoluble Table 1: Microorganisms and Hydrocarbon (organic compound) interaction. Compound Monoeyelie aromatic hydro carbons, benzene, toluene, ethyl benzene and xylene ,phenol compounds Microorganisms Penicillium chrysogenum P. alcaligenes, P. mendocina and P. putida, P. veronit, Achromobacter, Flavobacterium, Acinetobacter Pseudomonas putida Phanerochaete chrysosporium A. niger, A. fumigatus, F. solani and P. funiculosum bCoprinellus radians Alcaligenes odorans, Bacillus subtilis, Corynebacterium propinquum, Pseudomonas aeruginosa Tyromyces palustris, Gloeophyllum trabeum, Trametes versicolor Candida viswanathii Cyanobacteria, green algae and diatoms and Bacillus licheniformis Acinetobacter sp., Pseudomonas sp., Ralstonia sp. and Microbacterium sp, Gleophyllum striatum Acinetobacter sp., Pseudomonas sp., Ralstonia sp. and Microbacterium sp, Gleophyllum striatum ‘Acinetobacter sp., Pseudomonas sp., Ralstonia sp. and Microbacterium sp, Gleophyllum striatum Petrol and diesel polycyclic aromatic hydrocarbons toluene Monocyclic aromatic hydrocarbons, e.g. benzene and xylene. Biphenyl and triphenylmethane Hydrocarbon PAHs, methylnaphthalenes, and dibenzofurans Phenol Hydrocarbons Phenanthrene, benzopyrene ‘Naphthalene aromatic hydrocarbons striatum Pyrene, anthracene, 9- metil anthracene, Dibenzothiophene Lignin peroxidasse aromatic hydrocarbons striatum Pyrene, anthracene, 9- metil anthracene, Dibenzothiophene Lignin peroxidase aromatic hydrocarbons striatum Pyrene, anthracene, 9- metil anthracene. Dibenzothiophene Lignin peroxidase rable 2: Grou ’Ps of microorganisms important for oil bioremediation. Microorganisms c id ‘ompoun, Fusarium sp. 7 il oi Alcaligenes odorans, Bacillus subtilis, C inn, tonal nme * Bacillus cereus A diesel oil Aspergillus niger, Candida gl ; Candida glaby m Saccharomyces cerevisiae ae Ro te seus oll B. brevis, P. aeruginosa KH¢ a 16, B. licheniformis and B. enue oil Pseudomonas aeruginosa, P. putie Eivedanen . putida, Arthobacter sp and diesel oil Pseudomonas cepacia, Bacillus cereus, Bacillus coagulans, diesel oil, rude oil Citrobacter koseri and Serratia ficaria ‘Table 3: Representative examples of most dominate microorganisms in the involvement of dyes bioremediation Microorganisms ‘Compound B. subtilis strain NAP1, NAP2, NAPA oil-based based paints industrial dyes ‘Myrothecium roridum IM 6482 Pyenoporus sanguineous, Phanerochaete chrysosporium and Trametes - is industrial dyes trogit Penicillium ochrochloron “Micrococcus luteus, Listeria denitrificans and Nocardia atlantica eudomonas aeruginosa, Bacillus pumilus Textile Dye (Remazol Black B), azo dye Reactive Red HEB, RNB dye industrial dyes Textile Azo Dyes Bacillus spp. ETL-2012, Psi Sulfonated di- HKG212 Exiguobacterium indicum, Exiguobacterium aurantiacums, Bacillus azo dyes effluents cereus and Acinetobacter baumanit Bacillus firmus, Bacillus macerans, Staphylococeus aureus and vat dyes, Textile effluents Klebsiella oxytoca Table 4: Microorganisms serve for utilizing heavy metals. Compound Microorganisms Heavy metals, lead, mercury and nickel Saccharomyces cerevisiae Cunninghamella elegans Pseudomonas fluorescensand Pseudomonas aeruginosa Heavy metals Fe 2+, Zn2+, Pb2+, Mn2+ and Cu? Lysinibacillus sphaericusCBAMS cobalt, copper, chromium and lead Microbacterium profundi strain Sbh49 Fe Terichoderma sp., Mic P.. Microsporum sp., Cl a colin, feaaigee ’sporum sp., Cladosporium sp. ¢ Fe (II), U(VD. Bacillus safensis (IX 126862) strain (PB-S and RSA-4) Cadmium Pseudomonas aeruginosa, Aeromonas sp. U, Cu, Ni, Cr Aerococcussp., Rhodopseudomonas palustris Pb, Cr, Cd Table S: Potential biological agents for pesticides. Microorganisms Compound Bacillus, Staphylococcus Endosulfan Enterobacter Chlorpyrifos Pseudomonas putida, Acinetobacter sp, Arthrobacter sp. fll ZOE MC, le Ure Deee SPB, malation Acenetobacior sp., Pseudomonas sp., Enterobacter sp. and Phos chlorpyrifos and methyl parathion BACTERIAL DEGRADATION ia with the ability to degrade Several bacteria are known to feed exclusively on hydrocarbons. Bacteri hydrocarbons are named hydrocarbon-degrading bacteria. Biodegradation of hydrocarbons can occur under aerobic and anaerobic conditions, it is the case for the nitrate reducing bacterial strains Pseudomonas sp. and ‘ted from petroleum contaminated soil. Others are as follows: Bacillus, Corynebacterium, Brevibacillus sp. isolat Staphylococcus, Streptococcus, Shigella, Alealigenes, Acinetobacter, Escherichia, Klebsiella and Enterobacter. Bacterial strains that are able to degrade aromatic hydrocarbons have been repeatedly isolated, mainly from soil. These are usually gram negative bacteria, most of them belong to the genus Pseudomonas. Although many bacteria are able to metabolize organic pollutants, a single bacterium does not possess the enzymatic capability to degrade all or even most of the organic compounds in a polluted soil. Mixed microbial 1 biodegradative potential because the genetic information of more than one onganism is necessary to degrade the complex mixtures of organic compounds present in contaminated areas. Both, anaerobic and aerobic bacteria are capable of biotransforming PCBs. Higher chlorinated PCBs are ted to reductive dehalogenation by anaerobic microorganisms. Lower chlorinated biphenyls are oxidized search on aerobic bacteria isolated so far has mainly focused on Gram-negative strains era Pseudomonas, Burkholderia, Ralstonia, Achromobacter, Sphingomonas and several reports about PCB-degrading activity and characterization of the genes that are involved in PCB degradation indicated PCB-degrading potential of some Gram-positive strains as well (genera Rhodococcus, Janibacter, Bacillus, Paenibacillus and Microbacterium), Aerobie catabolic pathway for PCB degradation seems to be very similar for most of the bacteria and comprises four steps eatalysed by the enzymes, biphenyl dioxygenase (BphA), dihydrodiol dehydrogenase (BphB), 2, 3-dihydroxybihenyl dioxygenase (DHBD) (BphC) and hydrolase (BphD). Successful removal of pesticides by the addition of bacteria had been reported earlier for many compounds, trazine, Recent findings concerning pesticide degrading bacteria include the Chlorpyrifos degrading Providencia stuartii isolated from agricultural soil and isolates Bacillus, Staphylococcus and vated and uncultivated soil able to degrade dichlorodiphenyltrichloroethane} communities have the most powerful subject by aerobic bacteria. Re: belonging to the gent ‘Comamonas. However, including a bacterium, Stenotrophomonas trom culti (DD). ade azo dyes under aerobic and anaerobic conditions have en extensively reported, Based on the available literature, microbial decolourization of azo dyes is more effective under anaerobic conditions. On the other hand, these conditions lead to aromatic amine formation, and these are mutagenic and toxic to humans requiring 4 subsequent oxidative (aerobic) stage for their degradation. For eg, the mixed consortia of bacteria consisti f Proteus sp., Pseudomonas sp, and Enterococews sp. can be sed in the biodegradation and decolorisation of dye. Heavy metals cannot be destroyed biologically (no'‘degradation”, change in the nuclear structure of the element, occurs) but are only transformed from one oxidation st «x to another. Microorganisms have developed the capabilities to protect themselves from heavy metal toxicity by various mechanisms, such as adsorption, uptake, methylation, oxidation and reduction. Reduction of metals can occur through dissimilatory metal reduction, where bacteria utilize metals as terminal electron acceptors for anaerobic respiration. In addition, bacteria may possess reduction mechanisms that are not coupled to respiration, but instead are thought to impart metal resistance. For example, reduction of Cr(VI) to Cx(IT1) under aerobic or anaerobic conditions, reduction of Se(V1) to elemental Se, reduction of U(VI) to UCI) and reduction of Hg(Il) to Hg(0). Microbial methylation play an important role in heavy metals bioremediation, because methylated compounds are frequently volatile. For example, Mercury, Hg(ll) can be biomethylated by a number of different bacterial species Alcaligenes faecalis, Bacillus pumilus, Bacillus s ip, P. aeruginosa and Brevibacterium /odinium to gaseous methyl mercury. In addition to redox conversions and methylation reactions, acidophilic iron bacteria tke slcidthiobacilus ferrooxidans and sulfur oxidizing bacteria are able to leach high concentrations of As, Ca, Cu, Co and Zn from contaminated soils. On the other hand, metals can be precipitated as insoluble sulfides indirectly by the metabolic activity of sulphate reducing bacteria. Sulphate reducing bacteria are anaerobic heterotrophs utilizing a range of organic substrates with SOs” as the terminal electron acceptor. PLANT GROWTH PROMOTING _RHIZOBACTERIA _(PGPR) AND PLANT GROWTH PROMOTING BACTERIA (PGPB) DEGRADATION at occur naturally in plants) Plant assoviated bacteria, such as endophytic bacteria (non-pathogenic bacteria th and thizospherie bacteria (bacteria that ive on and near the roots of plants), have been shown to contribute to biodegradation of toxic organic compounds in contaminated soil and could have potential for improving phytoremediation. Plant growth promoting rhizobacteria (PGPR) are naturally occurring soil bacteria that aggressively colonize plant roots and benefit plants by providing growth promotion, Some plants can release structural analogs of PAHs such as phenols, to promote the growth of hydrocarbon degrading microbes and their degradation on PAHs. For such plant/microbe systems, an important class of bacteria, Pseudomonas spp. have PGPR activity and hydrocarbon degrading capacity. Furthermore, the thizosphere of vegetation in diversity of population of PAH-degrading bacteria, among which two Lysini contaminated field contains higher bacillus strains were isolated. Culturable PCB degraders were also associated with both the rhizosphere and root zone of mature trees growing naturally in a contaminated sit, they were identified as members of the genus Rhodococcus, Luteibacter and Williamsia, which suggest that biostimulation through rhizoremediation is @ promising strategy for enhancing PCB degradation in situ, Also, the free living nitrogen fixer

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