Environmental

Biotechnology
Sarah Yorba
Xuan Jing Teoh
Oscar Detrinidad
Karen Vore
 4 Major Categories

1) BIOMAINTENANCE gene flow between fragmenting populations

2) BIOREMEDIATION The use of living organisms to clean up

contaminated soil or water . The use of plants to clean up the environment,
known as phytoremediation, is also considered a type of bioremediation.

3) BIOWASTE MINIMIZATION The use of biotech techniques and

products that are both environmentally sound and economically acceptable to
prevent adverse environmental effects (think "green" detergents)

4) BIOSENSORS Environmental monitoring or diagnostics using indicator

organisms for early detection.
 What is Bioremediaton?

Any process that uses microorganisms,

fungi, green plants or their enzymes to
return the natural environment altered
by contaminants to its original
condition
 Contaminants
Nonhalogenated volatile organic compounds (VOCs).
Halogenated volatile organic compounds.
Nonhalogenated semivolatile organic compounds
(SVOCs).
Halogenated semivolatile organic compounds
Fuels.
Inorganics.
Radionuclides.
Explosives.
Microorganisms need

Oxygen

Nutrients

Moisture

Optimal temperature and pH

Processes used include
In situ
Ex situ
(both can involve biological, physical, chemical or thermal treatments)

Natural Attenuation
Enhanced Attenuation
(bioaugmentation or biostimulation)
In Situ Biological Treatments

Bioventing

Enhanced Bioremediation

Phytoremediation
Bioventing and Air Sparging

Oxygen is forced into contaminated soil or

groundwater converting organic materials to carbon
dioxide, water and microbial cell mass. Source of
oxygen can be hydrogen peroxide.
Enhanced Bioremediation

Involves the addition of microorganisms (e.g., fungi, bacteria, and

other microbes) or nutrients (e.g. oxygen, nitrates) to the subsurface
environment to accelerate the natural biodegradation process.

Both aerobic and anaerobic methods are used.

 Enhanced Bioremediation

Soil
Aerobic: Percolation or injection of water mixed with nutrients and saturated
with dissolved oxygen.

Anaerobic: Methane, Sulfate, Nitrate commonly used. The organic

contaminants will be ultimately metabolized to methane, resulting in limited
amounts of carbon dioxide, and trace amounts of hydrogen gas.
 Enhanced Bioremediation

Groundwater
Aerobic: Oxygen enhancement can be achieved by either sparging air below
the water table or circulating hydrogen peroxide throughout the contaminated
ground water zone.
Anaerobic: Nitrate is circulated throughout the ground water contamination
zone to enhance bioremediation. Solid-phase peroxide products (e.g., oxygen
releasing compound (ORC)) can also be used for oxygen enhancement and to
increase the rate of biodegradation. Oxygen is the main electron acceptor for
aerobic bioremediation. Nitrate serves as an alternative electron acceptor
under anoxic conditions.
 Limitations
Long-term technology which may take several years for cleanup.

Low temperature slows the remediation process.

Under anaerobic conditions, contaminants may be degraded to a product that is more hazardous
than the original contaminant. For example, trichloroethylene (TCE) anaerobically biodegrades to
the persistent and more toxic vinyl chloride. Under anaerobic conditions, contaminants may be
degraded to a product that is more hazardous than the original contaminant. For example,
trichloroethylene (TCE) anaerobically biodegrades to the persistent and more toxic vinyl chloride.

High concentrations of hydrogen peroxide in groundwater inhibit the activity of microorganisms.

Amended oxygen can be consumed very rapidly near the injection well, which creates two
significant problems: biological growth can be limited to the region near the injection well,
limiting adequate contamination/microorganism contact throughout the contaminated zone; and
bio-fouling of wells can retard the input of nutrients.

Bioremediation is not well suited for soils with low permeability (e.g., fine clays). High
permeability is required to allow the nutrients to reach the indigenous microorganisms.
 Limitations
It is possible that the subsurface injection of gases below the water table can induce groundwater
flow. It may be necessary to use a pump-and-treat system in conjunction with gas injection for
hydraulic control.

The circulation of water-based solutions through the soil may increase contaminant mobility and
necessitate treatment of underlying groundwater. If the process is enhancing groundwater
bioremediation, a groundwater circulation system must be created so that contaminants do not
escape from zones of active biodegradation. See description of Circulating Groundwater Wells.

Nitrate injection to groundwater is of concern because nitrate is a regulated compound. Bio-

augmentation using non-native microorganisms is also controversial.

Very high contaminant concentrations may be toxic to microorganisms.

Safety precautions must be used when handling hydrogen peroxide.

Because gaseous injection increases pressure in the soil, vapors can build up in building
basements.
 Genetically Engineered
Microorganisms
Microorganism Modification Contaminants

Pseudomonas sp. B13 Pathway mono/dichlorobenzoates

P. Putida Pathway 4-ethylbenzoate

P. putida KT2442 Pathway toluene/benzoate

Pseudomonas sp. FR1 Pathway chloro-, methylbenzoates

C. testosteroni VP44 Substrate o-, p-monochlorobiphenyls

specificity

Pseudomonas sp. LB400 Substrate PCB

Specificity

E. coli JM109(pSHF1003) Substrate PCB, benzene, toluene

Specificity

P. Pseudoalcaligenes KF707-D2 Substrate TCE, toluene, benzene

Specificity

E. coli FM5/pKY287 Regulation TCE, toluene

 Phytoremediation:
the use of plants to extract, sequester,
detoxify, and/or hyperaccumulate toxic
pollutants from soil, water, and air.
This process can be exponentiated through
the application of biotechnology and the
expression of foreign genes by plants.
 Currently
Excavation and
relocation-
expensive!

natural attenuation- can take thousands of years!

 2 Major Pollutants
Organic pollutants: Elemental pollutants:
Carbon-containing heavy metals and
compounds like benzene; metalloids
leftovers from industrial
era.
How it Works
 METHODS
Phytoextraction: Heavy metals-
stored in the roots, stems, or leaves

Phytodegradation: organic toxins-

changed into less harmful chemicals within the plant
using enzymes

Phytovolatilization: soluble contaminants-

changed into gases that are released into the air as the
plant breathes
 And then
they are harvested and destroyed (burned)
recycled if metals stored in the plants can be
reused
trees are left to grow and are not harvested.
 Possible Insert-Gene Examples:

increase the solubility of metals in the soil

surrounding the roots
transport proteins that move metals into root
cells
further transport to other parts of the plant
oxidation or reduction
degradation and minerilization
sequestration and binding
 Plant examples:
researchers at the University of Georgia genetically modified yellow poplar
trees with a gene from a mercury-resistant bacteria
Leon Kochia and molecular biologist David

Garvin study wheat plants of various

genotypes being studied for aluminum
tolerance.
 Arabidopsis genes

700 genes encode proteins that have the capacity to act directly
on environmental pollutants
8090% of all the phytoremediation genes that could be used in
phytoremediation applications
can accumulate up to 30,000 ppm zinc and 1,500 ppm cadmium.
A normal plant can be poisoned
with as little as 1,000 ppm of zinc or 20 to
50 ppm of cadmium in its shoots
Zn transport gene cloned,
led to regulation discovery
 Regulation
In normal plants, toxin uptake regulated by
current levels of element in plant.
In hyperaccumulators, transporter genes are
maximally active at all timesindependent of
plant levels
 Limitations
Plants can only clean up chemicals as deep as
their roots can grow (3-6ft for shrubs, 10-15ft
for trees)
Plants can be eaten or destroyed by weather
Clean-up can take 3+ years
Plants must be watered, fertilized, and
monitored
 Safety
EPA tests the plants and air to make sure that
the plants do not release harmful gases into the
air
insects and small animals may eat the plants,
moving up the food chain.
can help keep harmful chemicals from moving
from a polluted site to other areas
(stabilization).
Environmental Problems

pollution of local surface

water and ground water
with phosphorus
high phosphorus content
lead to the growth of
plant and algal
death of fish and other
beneficial aquatic
organisms
Enviropigs

Transgenic pigs

produce low-phosphorus
manure
 Purpose of designing Enviropigs

Enviropigs manure contain 75% less

phosphorus than average swine

Can deplete oxygen level in water downstream

 Developed at University of Guelph in Ontario
Introduced to the world in August
Dr. John Phillips the leader researcher in the
Enviropigs project.
 DNA-modified Piglets

Jacques

Gordie

Wayne

*named after Canadian Hockey Players

 Characteristic of Enviropigs

produces the enzyme phytase

synthesized phytase in larger quantities in the
salivary glands
produce sufficient phytase to digest practically
all of the phytate in a cereal grain diet.
 How the EnviropigTM Works
Phytase produced in the
salivary glands and
secreted in the saliva
Pancreas Cecum
Liver

Stomach
Indigestible Manure
Phytate
phosphorus low in
Phytase Small
in the diet Intestine Phosphor
PHOSPHATE Colon us
Phytate Phosphate ABSORPTION

Phytate in a non-Transgenic pig

Phytate in the Enviropig
Process of modifying pigs digestives
abilities
Combined bacterial gene which make an enzyme
that break down the form of phosphorus with a
mouse gene
This combination will cause the enzyme to be
secreted from animals mouth.
The composition gene was then injected into
one-celled pig embryos that were subsequently
surgically implanted into a surrogate mother.
Plant Biotechnology
And the Environment
 Environmental Effect of Pesticide
and Herbicide
Pesticide Drift
Air Pollution

Water Pollution

Soil Pollution

Organic Pollutant
 Causes of Environmental
Pollution
Chemicals
Pesticide
Herbicide

Energy
Noise
Heat

Radiation
 How Plant Biotechnology
develops pathogen resistant
plants
Insecticidal Protoxin

Alpha-amglase inhibitors

Protease inhibitors
 Reduction of pesticide due to
pathogen resistant plants
Reduced 30% a year for all pesticide use in U.S.A
for:
Maize
Barley
Alfafa

With the finding of new pest resistant genes

target pest control is greater than 80%. This
means less pesticide/herbicide use.
 Reduction of pesticide due to pest
resistant
Bt Crops
Bt maize

Bt cotton

In the U.S.A insecticide use was

reduced by 2.2 sprays. Therefore,
decreasing use of pesticide/ herbicide.
 Pests will develop resistant to
transgenic pesticidel crops
Trait for resistance to a specific insecticide
produced by a plant is already present.
Increased mating of the resistant individuals
with other resistant individuals product larger
numbers of resistant individuals
What this mean is that scientist most discover
new pest resistant gene to use in plants in order
to not use pesticide/herbicide as often as in the
past.
 References
Bioremediation. Wikipedia, The Free Encyclopedia. 28 Nov. 2008. http://en.wikipedia.org/wiki/Bioremediation
Enhanced Bioremediation. Center for Public Environmental Oversight. 28 Nov. 2008.
http://www.cpeo.org/techtree/ttdescript/ensolmx.htm
Contaminant Perspectives. Federal Remediation Technologies Roundtable. 28 Nov. 2008.
http://www.frtr.gov/matrix2/section2/2_intro.html
In Situ Biological Treatment for Soil, Sediment, Bedrock and Sludge. Federal Remediation Technologies Roundtable. 28 Nov.
2008. http://www.frtr.gov/matrix2/section3/3_1.html
In Situ Biological Treatment for Ground Water, Surface Water, and Leachate. Federal Remediation Technologies Roundtable. 1
Dec. 2008. http://www.frtr.gov/matrix2/section3/3_9.html
Fu-Min Menn, James P. Easter, Gary S. Sayler, 21 Genetically Engineered Microorganisms and Bioremediation. Environmental
Information System Centre. 1 Dec. 2008. http://www.envismadrasuniv.org/pdf/Genetically%20Engineered%20Microorganisms.pdf
Wendy Ann Peer, Ivan R. Baxter, Elizabeth L. Richards, John L. Freeman, Angus S. Murphy, Phytoremediation and
hyperaccumulator plants. Purdue University. www.hort.purdue.edu/hort/research/murphy/pdfs/metals11.pdf
Phytoremediation: Using Plants To Clean Up Soils. USDA website. http://www.ars.usda.gov/is/AR/archive/jun00/soil0600.htm
A Citizens Guide to Phytoremediation. EPA website. http://clu-in.org/download/citizens/citphyto.pdf
Phytoremediation. BioBasics. http://www.biobasics.gc.ca/english/View.asp?x=742
Hessler, Kristen. Ethics Case Study: Enviropigs. Office of Biotechnology, 2003. Iowa State University. 18 Nov. 2008
http://www.bioethics.iastate.edu/classroom/enviropigs.html
Forsberg, Cecil W. Guelph Transgenic Pig Research Program. n.d. University of Guelph, Guelph, Ont. 18 Nov. 2008
http://www.uoguelph.ca/enviropig/#healthy