biotechnological applications for environmental waste management
TRANSCRIPT
Biotechnological applications for environmental waste management
Natural Xenobiotic compounds
Emerging toxicants containing waste
Feb. 17, 2014
What is environment? Why environmental problems?
What are remedial methods? Technology development?
Major environmental issueImpact of green house gases
Formation of dioxin-like compounds in the environments
Important environmental problems
• 1. Global warming- GHG (CO2, CH4, N2O)
• 2. Energy problem- Bioethanol, biodiesel, Hydrogen how?-a substitute-
• 3. Water contaminants/toxicants/eutrophication • 4. Soil degradation/solid waste generation
• 5. Air pollutants
3. Molecular Biology-Catabolic
Enzymes & genes
1. Chemistry-
Extraction &
analysis
of pollutants
2.
Microorganism
& Microbial
Ecology
5. T
oxic
olog
y-To
xico
geno
mic
s&
det
oxifi
catio
nR
epor
ter
Gen
e
4. E
ngin
eerin
gM
olec
ular
Pr
obes
& b
iose
-ns
or
Environment
Air,
Water and soil
-Natural
Xenobiotic
compounds
T
BioChemicalsProcess
Engineering
Biotechnology
Bio Environment&
Engineering(Environmental
Biotechnology)
4.2 Microbial Bioremediation
in situ & ex situ
4.1 PhytoremediationCaviation methods
Bioremediation- A potential approach for clean and green environment
5.Bioproducts &
biomaterials
6. System approach
Climate Climate/meteorology
Origin of Earth and Environment
The Universe created by colossal explosion that we now refer to as the Big Bang and Planets of the solar system
The Earth and Environment
The earth
The ocean
Climatic changes on the Earth
Movement of air during rotation of the Earth and formation of cells
PHYSICAL FACTORS AND BIOTIC & ABIOTIC MATERIALS IN ENVIRONMENT
Origin of life on the Earth
Classification of living organisms
Biodiversity
Classification of microorganisms
Classification of plants and animals
Biomolecules in organisms• It is organic compound composed of carbon, hydrogen, oxygen, nitrogen,
sulfur, phosphorus and sometimes some other elements.
• Different types of biomolecules are:• A. Small molecules mainly include molecules like:-• Lipids such as phospholipids, glycolipids, sterols, and glycerolipids: -• Carbohydrates- provide energy and act as energy storage molecules.• Vitamins-survival and health of organisms.• Hormones, neurotransmitters and metabolites: - metabolic processes and
functions.
• B. Monomers include:-• Amino acids: - building blocks of proteins function as genetic code and as
biomolecules, that assist in other processes such as lipid transport.• Nucleotides: - Chemical energy (ATP,GTP), assist in cellular signaling, and
enzymatic reactions (coenzyme A, flavin adenine dinucleotide, flavin mononucleotide, nicotinamide adenine dinucleotide phosphate etc ).
• Monosaccharide: - provides energy and are the building blocks of polysaccharides.
Emergence of man and social environment
• Understanding Human Behavior and the Social Environment
• Natural Resources:Air, water, soil, minerals etc.
• Industrial Revolution-1760-1850 onwards
Environmental degradation • The ten threats identified in 2004 by the High Level
Threat Panel of the United Nations are these:• Poverty• Infectious disease• Environmental degradation• Inter-state war• Civil war• Genocide• Other Atrocities (e.g., trade in women and children for
sexual slavery, or kidnapping for body parts)• Weapons of mass destruction (nuclear proliferation,
chemical weapon proliferation, biological weapon proliferation)
• Terrorism• Transnational organized crime
Major environmental issues before us
Pulp and paper mill effluent Molasses from sugar cane mill for distillation
17 million gallon oil spill under the Greenpoint section of Brooklyn
Waste dumping grounds in Delhi
Petroleum waste
Contd.
Emerging industrial pollutantsIndustrial sources• Pulp and paper
industry lignosulphonic acid,
chlorinated resin acid, chlorinated
phenolsdioxins, dibenzofuran,
biphenychlorinated hydrocarbonDistillery industry melanoidinsTannery industryChlorinated phenolics,PCPs, chromiumMunicipalPlastic, dioxins, antibiotic
etcTransportMetals, organics
Incineration and plastics etc.
Pops in distillery effluent Pops in pulp & paper effluent
Pops in tannery effluent
Pops in municipal sludge
Fate of Organic Compounds in the Environment
AIR
Water Soil
ENVIRONMENTAL POLLUTANTS
Major conferences and meetings• United Nations Conference on the Human Environment-
Sweden in 1972: Declaration containing 26 principles concerning the environment and development 6. Pollution must not exceed the environment’s capacity to clean itself19. Environmental education is essential20. Environmental research must be promoted, particularly in developing countries
The United Nations Conference on Environment and development (UNCED)- Rio Summit and- Earth Summit
United Nations Framework Convention on Climate Change-Kyoto Protocol-reduce emissions of greenhouse gases
In Doha, Qatar, on 8 December 2012, the "Doha Amendment to the Kyoto Protocol
Global warming gases in the environment
The potential mechanisms that regulate the responses of GHGs (CO2, CH4 and N2O)
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Production and consumption to elevated N (ANPP, aboveground net primary productivity; BNPP, belowground net primary productivity; SOC, soil organic carbon; DOC, dissolved organic carbon; DIN, dissolved inorganic nitrogen; DON, dissolved organic nitrogen).
Climate change and BiodiversityRole of organisms- autotrophic & chemoautotrophic in CO2 mitigation
Carbonic anhydrase Biosurfactants Bioscrubbers for CO2 sequestration
Solid waste generation from different sources
1. Garbage- putrescible, heating value 2. Rubbish- Non putrescible, heating value
3. Pathological4. Industrial Municipal waste5. Agriculture waste6. Medical waste7. Electronic waste
Biodegradable Non biodegradable waste
Natural waste Xenobiotic Hazardous- ignitable (i.e. flammable), oxidizing, corrosivity, toxic Radioactive, eco-toxic, explosive
Non-hazardous waste
Biogas
Waste water treatment options
Primary treatment
ScreeningGrit removalEqualizationStorageGrindersFlocculationSedimetationFloatationCoagulation
Secondary treatment
Aerobic Anaerobic
Tertiarytreatment
Chemical oxidationFiltrationCarbon adsorptionOsmosisElectrolysisCavitationsPhotodegradation
Activated sludge processTricking filterFixed film reactorRotating reactorStabilization pond
Upflow anaerobic sludge blanket reactorAn. Fludized bed reactorAnaerobic lagoonsAn. Contact reactorAn baffled reactors
Origin of different types of chemical compounds in the environment and their fate
Significance of lignocellulosics• Total forest cover 3870 million hectares or 30% of the earth’s
land area.• 50% 0f all biomass with an estimated annual production of 50
billion tons.
• Half of the residues remain unused while some are used as material and energy-green manure and feed for low producing ruminants.
• Major substrate for food, feed, energy, and other commercial items.
• Degrading enzymes have potency for fuel, chemicals, food, brewery and wine, animal feed, textile and laundry, pulp and paper, agriculture and pharmaceuticals.
• Unused biomass is major source of “waste”- pose an environmental pollution problem.
Lignocellulosic ecosystem : cellulolytic, hemicellulolytic and liglinolytic strains
Structure of lignocellulose• Cellulose : Made up of linear chains of β-1,4-linked
D-glucose residues.
• Hemicellulose : Made up of branched heteroglycans with a backbone of β-1,4-linked D-xylopyranosyl residues with branches of α-1,3-linked L-arabinofuranosyl and α-1,2-linked 4-O-methyl-glucoronic acid residues.
• Lignins is heterogeneous, three dimensional polymer composed of oxyphenyl propanoid units connected by c-c and c-o-c linkages. It is formed by random coupling of coniferyl alcohol, sinapyl alcohol and p-coumaryl alcohol.
Lignocellulosic components and its importance as biomaterials
Lignocellulose
Cellulose Hemicellulose
LigninVanillin
Gallic acidPhamaceuticals
HerbicidesAntifoming agents
House hold productsPulp
GlucoseCellulosederivatives
Fuel
Feed and commercial
items
FurfuralsXylosepulp
Single cell proteinsXylitol
Degradation of cellulose by enzyme cellulase
Pre hydrolysis: Acid, Alkali, ammoniaEnzymes: Thermolhilic, alkalophilic, multiplicityProducts-fuel. feed, food, commercial productsBiofuels
Applications: Pulp, industries, food, feed, fuel etc.
Generalized mechanism of enzymatic cellulose hydrolysis
Problems:1. End product inhibitions
Biotechnology1. Mutants 2.Protoplast fusion3. Genetic engineering 4. More enzyme5. Protein engineering 6. Cellulosome-
multicomponents enzyme system
Hemicellulose and degradation- Enzyme xylanase
•HC is homo and heteropolymer•AnhydroB-(1,4)D-xylopyrannose, mannopyranose, glucopyranose, galactopyranose•Monomer is D-Xylose
Applications1. Energy 2.Food & feed industries3. Pulp and paper- Biopulping & biobleaching4. Waste management5. Saccharifications of agrowaste6. Nutritional quality 7. Enhancing texture
Lignin structure and degradation
1. Prior 1970- no information for degradation2. 14C-labelled synthetic lignin3. Electron microscopy4. Lignin degrading fungi-White rot, soft rot, Brown rot, other5. Enzymes6. Physiological parameters-oxygen, nitrogen, carbon, temp. pH, nutrients
Fig. 3: The three common monolignols
Involvement of enzymes in degradation of lignin
1. Lignin peroxidase (LiP)Extracellular, H2O2 dependent, glycosylated hemprotein, MW 41-42 kDa, 2. Manganese peroxidase (MnP)Extracellular, H2O2 dependent, MnII-dependent, neutral carbohydrate, MW 41-45 kDa
3. LaccaseExtracellular, non-heme, copper containing
4. Other phenol-oxydizing enzymes
Applications1. Industrial, 2.Commercial, 3. house holds, 4. waste management
5. Glyoxal oxidaseSupport oxidative turn over of LiP and MnP reduction of O2 to H2O2 with oxidation of substrate
Bioethanol
1.Cellulases2.Xylanases3.Laccase-4.Lignin peroxidase &5.Manganese peroxidase
Biodegradation and bioconversion of lignocellulosic waste in the environment
Fermentation
Schematic diagram- ethanol production from sugarcane bagasse
Biotechnological innovations: biomaterials- biorefinery
• Screening for organisms with novel enzymes: enzyme evolution-random mutagenesis-recombination-selection-screening
• Strain improvement of existing industrial organisms and enzyme engineering
• Production and operation related factors-Process optimization– Substrate– Culture conditions– Recycling of enzymes– Redesigning of processes
– Process optimization models and soft wares
Strain improvement of existing industrial organisms and enzyme engineering
• Hyper producer organisms• Robust organisms
– Culture conditions: isolation of 1% strains-Great culture plate enigma
– Biomining through:• Genomics-complete blue print of the organism• Metagenomics-genomics with functional aspects at
community level– Necessity of discovering unique gene, cloning,
quantitative analysis, and expression
Process optimization Bioreactors
Liquid stateFermentation
-Homogeneous -Heterogeneous
Stirred tank reactorAir-lift or bubble-column
reactor
Solid state Fermentation
FlaskTray
Packed bedTunnelPaddle
Rotating drumTower
Batch
Continuous
Fed-batch
laboratory scalePilot scale
Industrial scale
Biofuel Production and integrated pollution control using microalgae
• Microalgal Farming and CO2Mitigation• Microalgal Farming using Wastewater• Microalgal Farming using Marine Microalgae
Possible routes to energy products
Basic overview of the pathway of carbon capture and lipid biosynthesis
Anthropogenic chemical compounds in environemnt
Persistent organic pollutants in environment• Wide distribution- POPs detected from soil, water, food
items, commercial products
• Sources- Mostly chlorinated organic compounds formed unintentionally- industries, commercial, agriculture, military, other human activities, and natural sources
• Insufficient data- No reliable data for their persistence in Indian environment- No management practices
• Problems in detection methods- Methods for detection and degradation not up to the mark.
• Highly toxic and recalcitrant- ultimate formation of- tetrachlorodibenzo-p-dioxin and furan-like compounds-complete physiological impairment.
• Tremendous scope for medical diagnostics and therapy and products.
• Therefore, methods & technology for detection, bioremediation and detoxification is required.
Degradation of aromatic compounds
Key component: POPs
Emerging environmental contaminant in present scenario
Classification of POPs• Dirty Dozen - UNEP Stockholm Convention on Persistent Organic Pollutants - 2001
aldrin dieldrin toxaphenechlordane endrin mirexpolychlorinated biphenyls heptachlor DDTpolychlorinated dibenzo-p-dioxins
polychlorinated dibenzofurans
hexachlorobenzene
• UNEP has added nine new chemicals (all are poly haloginated compounds) to the "dirty dozen" list of restricted or banned toxic chemicals in 2009.
• Some other organic pollutants that may be persistent or lead to formation of dioxin like compounds in the environment include:
Poly Aromatic Hydrocarbons
Aromatic amines Pyrethroids
Volatile Organic Compounds
Metabolites of VOCs Phthalates
Biomagnification>Biomagnification, also known as bioamplification, or biological magnification is the increase in concentration of a substance, such as the pesticide DDT, that occurs in a food chain as a consequence of: Food chain energetics >Low (or nonexistent) rate of excretion/degradation of the substance.
Persistence and detection of dioxin-like POPs• Dioxin detected from food items, human exposure, milk and its
products, environmental sources from US, Japan and EU countries.
• No reliable data from developing countries including India.
• Detection methods.
• Instrument development.
• Thermokinetic modelling, equilibrium modelling, statistical determinations and others.
• Field validation.
• Laboratory
Biodegradation strategies for removal of organic compounds in environment
• Possible use of biodegradation processes-Indigenous microorganisms
-Genetically modified microorganism
-Continuous enrichment of microorganism
#Culture dependent and culture independent microorganisms-Metagenomic approach
Fate of organic compounds in the uptake into the cells and degradation, assimilation and
mineralization
Catabolic gene in degradation of alkanesDegradation of methane
Remediation of POPs in waste sites
What is Bioremediation?• Bio = living• Remediate = to bring the sites and affairs • into the original states
• Bioremediation can be defined as any process that uses microorganisms, green plants or their enzymes to return the environment altered by contaminants to its original condition.
• Bioremediation technology using microorganisms was reportedly invented by George M. Robinson.
• Use of biological sciences and technology for metals and organic compounds remediation.
Potential alternative for conservation and management of environment
Bio = living Remediate = to bring the sites and affairs into the original states
Bioremediation can be defined as any process that uses microorganisms, fungi, green plants or their enzymes to
return the environment altered by contaminants to its original condition.
BIOAUGUMENTATIONBIOAUGUMENTATION
BIOSTIMULATIONBIOSTIMULATION
Enzymatic methodsEnzymatic methods
Ex situ BioremediationEx situ Bioremediation
In situ Bioremediation In situ Bioremediation
Bioremediation
Importance of soil–plant–microbial interactions in bioremediation
Soil–plant–microbial interactions in remediation of pollutants in environment
Biocolloid formation in metal bioremediationColloidal aggregation–flocculation or attachment to inorganic and organic particles in water can lead to settling and removal of metals from the water column to the bottom sediment
Technologies in BioremediationEx situ bioremediation• Electro kinetically enhanced remediation• Soil Washing• Soil mound Bioxidation ProcessDispersing by Chemical Reaction• Biocolloid formation• Bioreactors• Land Treatment• Composting• Lagoons (aerobic/ anaerobic)• Partial peroxidation
In situ bioremediation• Bioventing• Bioslurping• Biopiling
Limitation of in-situ removed by Enhancement of Bioremediation Use of microorganisms to degrade contaminants in saturated soils and groundwater obtaining harmless chemicals as end products
Biosafety assessment of leachate after biological treatments
EstrogenicityGenotoxicityCytotoxicity
MTT Assay Comet Assay E-Screen Assay
(Nwagbara et al. 2007)
(Singh et al. 1988)
(Vanparys et al. 2006)
References:1) Nwagbara O, Darling-Reed SF, Tucker A, Harris C, Abazinge M, Thomas RD and Gragg RD. 2007. Induction of cell death, DNA strand breaks, and cell cycle arrest in DU145 human prostate carcinoma cell line by benzo[a]pyrene and benzo[a]pyrene-7,8-diol-9,10-epoxide. International Journal of Environmental Research and Public Health.4: 10–14. 2) Singh NP, McCoy MT, Tice RR and Schneider EL. 1988.A simple technique for quantitation of low levels of DNA damage in individual cells.Experimental Cell Research.175: 184-191.3) Vanparys C, Maras M, Lenjou M, Robbens J,Van Bockstaele D and Blust R. 2006.Flow cytometric cell cycle analysis allows for rapid screening of estrogenicity in MCF-7 breast cancer cells. Toxicology in Vitro.20:1238–1248.
*Huh 7 cell line is used for evaluating cytotoxicity and genotoxicity as hepatocytes express many nuclear receptor proteins that regulate the expression of xenobiotic metabolizing enzymes like CYP 1A1.*An estrogen receptive cell line MCF 7 is used for E-Screen assay.
Molecular Probes for tracking
Miniaturized ecogenomic sensors to measure microbial activity-carbon sequestration
• The sensors could be installed into advanced ocean observatories to monitor DNA and RNA from diverse microbial communities.
• Subsystems for monitoring, data management and communication, and data modelling would be incorporated for data contextualization.
• The sensors would report to a worldwide network of laboratories in real time by satellite telemetry.
• Culturable and nonculturable (metagenomics) bacteria for degradation of organic compounds & carbon concentrating mechanisms and value added products.
System biology approaches
The four-step paradigm for metabolic systems biology
Conclusion• POP/ DF and its congeners are difficult to
detect in the environment.
• Degradation of POP/DF in several steps by formation of intermediary metabolites.
• Degrading genes are present in various locations.
• Bioremediation difficult.
• Bioassay methods are useful which may be optimized and developed.
• System approach is recent days methods.
• Thanks