Presentation on theme: "Bioremediation methods and applications"— Presentation transcript:
1Bioremediation methods and applications ByDr. Ahmed Z. Abdel AzeizCollege of BiotechnologyMisr University for Science and TechnologyTel:
2Contents: 1- Definitions. 2-Bioremediation related topics Contents: 1- Definitions. 2-Bioremediation related topics. 3- Stages of a biodegradation study. 4- Cells and enzymes immobilization. 5- Phytoremediation. 6- Bioremediation of metals contaminated environment. 7- Biosurfactants 8- Bioremediation techniques: a) Soil bioremediation. b) Water and gasses bioremediation.
3DefinitionsBioremediation is any process that uses organisms (microorganism, algae and plant) or their enzymes to return the polluted environment to its original condition.Biodegradation is the use of these organisms in the degradation of different pollutants.Xenobiotic compounds are chemical compounds found in an organism but it are not normally produced or expected to be present in it.Cometabolism: in this process the microorganism produces an enzyme to utilizes its nutrients, but by chance this enzyme can degrade a pollutant.
4Bioremediation is a triple-corners process: SolidInorganicOrganicLiquidOrganismsPollutantsEnvironmentsGasMicroorganismsSoilWaterPlantsAirEnzymes
5Bioremediation related topics EnvironmentsOrganismsPollutantsBioremediation of metals polluted environmentPhytoremediationBioremediationEnzymes immobilizationCells immobilizationBioremediation techniquesBiosurfactantsEnzymes separation and identification
6Stages of a biodegradation study 1- Isolation of the microorganism2- Purification of the obtained isolates3- Identification of the microbial isolate4- Optimization of the biodegradation conditions5- Determination of the biodegradation efficiency6- Identification of the biodegradation products.7- Cell or enzyme immobilization.8- Enzyme identification.
71- Isolation of the microorganism - Isolation of bacteria and fungi is performed on Mineral Salt Medium (MSM) supplemented with the pollutant to be biodegraded as a sole source of carbon (enriched technique).- This medium can be used as liquid or solid.ControlSoil pollutant ControlIsolation of petroleum biodegrading bacteria from soil and petroleum pollution.Isolation of pesticide biodegrading bacteria from soil.
8BAFig. 2: Isolation of fungi (A) and bacteria (B) from the pollutant on solid MSM.The medium has a pH 7 is preferred for isolation of bacteria, while pH 4.5– 5.5 for fungi.
92- Purification of the obtained isolate This is simply performed by streaking of a lope of the MSM microbial culture or by pouring dishes technique.The used medium is solid MSM supplemented with the pollutant as a sole source of carbon.Purification by pouringPurification by streaking
103- Determination of biodegradation efficiency and products: 1- Spectrophotometer.2- HPLC3- GC/MS
114- Using the redox indicator 2,6-dichlorophenol indophenol (DCPIP). - The principle of this technique is that, during the microbial oxidation of the carbon source, electrons are transferred to electron acceptors such as O2, nitrates and sulphate.- DCPIP is an electron acceptor.- The efficiency is determined by observing the color change of DCPIP from blue (oxidized) to colorless (reduced).
12Cells and Enzymes immobilization: Immobilized enzymes (or cells) is defined as the enzyme that physically or chemically confined in defined materials with retention of its catalytic activity.The immobilization methods2- Cross-linking methods.3- Entrapping methods.1- Carrier-binding methods.1) Covalent binding methods1- lattice method.2- Microcapsule.2- Ionic binding methods.3- Membrane.3- Physical adsorption.4- Bio-specific binding methods.4- Reversed micelle.
13Materials used in the Carrier-binding methods 1- Polysaccharides: cellulose, dextran and agarose derivatives.2- Proteins: gelatin, albumin.3- Synthetic polymers: Polystyrene derivatives, ion exchange resins, polyurethane.4- Inorganic materials: glass, sand, ceramic and magnetite.
18PhytoremediationPhytoremediation is use of plants for accumulation, removal or conversion of pollutants.1-.PhytoremediationPhytostimulationPhytostabilizationPhytovolatilizationPhytotransformationPhytoextraction
20Approximately 400 plant species have been classified as hyperaccumulators of heavy metals, such as grasses, sunflower, corn, hemp, flax, alfalfa, tobacco, willow, Indian mustard, poplar, water hyacinth, etc.
21The root exudates of these plants play an important role in phytoremediation as it activate the surrounded microorganisms.Genetic engineering are used as in case of BT protein or insect pheromones producing plants to reduce the use of pesticides.
23Chemical structure of some siderophores pyridine-2,6-bis(thiocarboxylic acid)SEM images of selenium-siderophore complex formed in P. stutzeri culture filtrate.
24BiosurfactantsThe biosurfactants are chemical compounds characterized by hydrophobic and hydrophilic (non- polar and polar) regions in one molecule (amphipathic molecules).Biosurfactants from bacteria, cyanobacteria, fungi and yeast are classified into:Glycolipids.Lipopeptides.Phospholipids.Glycoproteins.Polymeric biosurfactants.
25Trehalose tetra ester from Arthrobacter sp. Chemical structure of some biosurfactantsGlucolipid from Alcalivorax sp.Trehalose tetra ester from Arthrobacter sp.Glucolipid from Alcaligens sp.
26Physiological roles of biosurfactants: 1- Increase the availability of hydrophopic compounds 2- Nutrient storage molecules. 3- Save the microbial cells from toxic substances. 4- Efflux of harmful compounds. 5- Extracellular and intracellular interactions such as quorum sensing and biofilm.
27Biosurfactant applications in bioremediation: The microbe may access a poorly water-soluble substrate that has been “pseudosolubilized” by the biosurfactant.Reduce the adsorption of the non-polar pollutants to the surface of soil particles.
28Bioremediation techniques: A) Soil bioremediation:(1) In-situ (without excavation).(2) Ex-situ (with excavation).Only ex-situ processes allow an efficient optimization of incubation parameters (biostimulation), including:pH,Aeration,Agitation,Moisteningnutrients,solvents or surfactants.In addition to addition of microorganisms (bioaugmentation).
301- High density poly ethylene (HDPE) 2- Sump pump to collect leachate3- Layer of pea gravel4- Layer of polluted soil to be treated5- Chopped alfalfa hay to retain moisture6- Wheels on sprinkler piping system7- Piping frame, aluminum or PVC pipes with frequent holes, sufficient to allow water, nutrients and bacteria to treat the land farm plot8- Flexible leachate collection hose9- Bypass valve that allows leachate to be circulated directly to water distribution tank,10- Recirculation hose11- Alken-Murray Bioactivator 2000, bioreactor unit12- Fresh water supply hoses13- Pumps for fresh water14- Treated water hose15- Water distribution tank16- Pump for distribution tank
31B- Water and gas bioremediation: Biofiltration is a process, in which, microorganisms supported on inert materials are used to degrade organic pollutants for air, gas and water bioremediation.Types of biofilters:1- Bioscrubbers.2- Biotrickling filters.3- Slow sand or carbon filters.
33Slow sand or carbon filters Slow sand or carbon filters work through the formation of a gelatinous layer (or biofilm layer) on the top few millimetres of the fine sand or carbon layer.This layer contains bacteria, fungi, protozoa, rotifera and a range of aquatic insect larvae (i.e. rotifers).
34Several research funding agency fund this type of applied research. Now each of us can start to work in a bioremediation research.