Bio-

remediation

wstafford@uwc.ac.za
Bioremediation
 The quality of life on Earth is linked inextricably to the
overall quality of the environment
 The contamination of soil and water with organic and
inorganic pollutants is of increasing concern and the
subject of legislation. These pollutants include complex
organic compounds, heavy metals, and natural products
such as oils and are derived from industrial processing,
deliberate release, and accidental release.

Bioremediation is defined as the process whereby

organic wastes are biologically degraded under
controlled conditions to an innocuous state, or to levels
below concentration limits established by regulatory
authorities.
 Bioremediation uses naturally occurring
bacteria and fungi or plants to degrade or
detoxify substances hazardous to human
health and/or the environment.
 The microorganisms may be indigenous to
a contaminated area and stimulated in
activity (biostimulation) or they may be
isolated from elsewhere and brought to the
contaminated site (bioaugmentation).
 In many cases the clean-up of contaminated sites has been carried
out using physical and chemical methods such as immobilization,
removal (dig and dump), thermal, and solvent treatments.
 Bioremediation is cheaper than the chemical and physical options,
and can deal with lower concentrations of contaminants more
effectively, although the process may take longer.
 The strategies for bioremediation in both soil and water can be as
follows.
 • Use the indigenous microbial population.
 • Encourage the indigenous population.
 • Bioaugmentation; the addition of adapted or designed
inoculants.
 • Addition of genetically modified micro-organisms.
 • Phytoremediation.
 Many thousands of synthetic organic (xenobiotic) compounds have been pro­duced
and many of these have found their way into the environment. Some of the most
commonly found are the pesticides (biocides), herbicides, and preser­vatives and
some of their structures are given in Fig. 5.2. Most biocides and herbicides are
released into the environment by direct use although some may be released during
manufacture, and spills do occur. The scale of herbicide production can be seen in
Table 5.2, where the herbicide atrazine is produced at a rate of 39 000 tonnes per
year. Other synthetic compounds such as poly-chlorobiphenyls (PCBs) are used in
hydraulic fluids, plasticizers, adhesives, and lubricants; flame retardants and dielectric
fluids in transformers are released during production, from spillage and disposal.
Another group of con­taminants found frequently in the environment are chlorinated
compounds such as trichloroethene, carbon tetrachloride, and pentachlorophenol,
which are used as solvents and for wood treatment. Other contaminants such as
dioxins and dibenzofurans can be formed during the combustion of polyaro-matic
hydrocarbons (PAHs) and when heating plant oils. So, depending on the properties,
such as solubility and the nature of their release, these contami­nants can be localized
or widespread.
 Many of the xenobiotic compounds released into the environment accumu­late
because they are only degraded very slowly and in some cases so slowly as to
render them effectively permanent. The half-lives (the time required to remove half of
the compound present) of some of the halogenated pesticides
Xenobiotic persistance
Recalcitrant chlorinated hydrocarbons
Agent orange (2,4-D and 2,4-T)
 The effect of the addition of another chlorine to
2,4-dichlorophenoxyacetic acid (2,4-D), a
biodegradable compound, forming 2,4,5-
trichlorophenoxyacetic acid (2,4,5-T), which is
recalcitrant. A mixture of 2,4-D and 2,4,5-T is
known as Agent Orange, and its persistence
was of great concern when it was used as a
defoliant in the Vietnam War.
 Caused long-term metabolic and neuroligical
effects in exposed humans
Oil spills
 As a result of the petroleum industry millions of
tons of these compounds enter the oceans every
year. Many hydrocarbons dissolve slowly in
water. Others such as the aromatic compounds
like benzene are more soluble, and these are
toxic to living cells.
 While accidental releases may contribute to only
a small percentage of the oil released into the
marine environment large accidental oil spills
receive much attention and evoke considerable
public concern because they can result in
contamination of ocean and shoreline
environments.
 oil spill!

The biggest spill ever occurred during the

1991 Persian Gulf war when about 240
million gallons spilled from oil terminals
and tankers off the coast of Saudi Arabia.
The Exxon Valdez accident at Bligh Reef
in 1989 discharged 40 million litres….
Bioremediation to the rescue?..
 Initial studies sowed that the number of oil degrading
microorganisms on oiled beaches in comparison with
untreated controls increased by as much as 10,000
times
 The biodegradation by indigenous microorganisms was
monitored by GC-MS, showed that that the microbial
population could rapidly biodegrade the aliphatic and
aromatic fractions of crude oil. The microbial community
decomposed dibenzothiphene, fluorenes, naphthalenes,
phenanthrene, and anthracene completely mineralized to
CO2 and H2O.
 Data also sowed that nitrate addition stimulated
biodegradation. Small scale in situ trials with Inipol EAP
22- an oleophilic microemulsion containing a solution of
urea in brine, encapsulated in oleic acid and lauryl
phosphate..
Initial experiments revealed visible
improvements after 10 days.
 More extensive beach plot experiments tested different
nutrient additions and an ecological monitoring program
was established (U.S. Congress 1989).
 Results showed that no detectable nutrients ended up in
the waters off the test site and there was no evidence of
eutrophication.
 The potential benefits of reducing wildlife exposure to oil
allowed the EPA to support a proposal for application of
nutrients to oil covered beaches (EPA 1989). By the end of
the summer of 1989, 100 km of shoreline were treated with
nutrient applications.
 The results of all tests showed that biodegradation can
be enhanced about two to three fold. This means that
an oil spill that would take five to ten years to degrade can
be degraded in as little as two to five years. This
acceleration of clean-up time, could give bioremediation
technology a bright future.
Biodegradable plastics?
 The use of long-lasting polymers for short-lived
applications is not entirely justified, especially when
increased concern exists about the preservation of living
systems.
 Most of today's plastics and synthetic polymers are
produced from petrochemicals. As conventional
plastics are persistent in the environment, improperly
disposed plastic materials are a significant source of
environmental pollution, potentially harming wildlife. (eg
1 in 30 cetacean carcasses had choked on plastic
debris). Plastics are also a costly in municipal waste
management.
 Pertochemical plastics take hundreds of years to break
down…
 but the time for biodegradable plastic to
be composted is 1 to 6 months :
Photobiodegradable plastics.
 Polymers that change structure when illuminated with UV
radiation, forming biodegradable materials.
Starch-linked biodegradable plastics.
 Starch has been incorporated into the structure of some
plastics, that can be microbially degraded.

Bacterial plastics.
 A number of microbes naturally produce biodegradable
polymers that are suitable for the plastics industry. For
example, the bacterium Alcaligenes eutrophus produces the
polyester poly-beta-hydroxybutyrate (PHB) as a storage
reserve for excess carbon. The monomeric unit of PHB is
beta-hydroxybutyrate.
 The strength, flexibility and crystallanity of polymers is
determined by the Type of repeating units and the media and
type of bacteria used to produce the polymer.
Why isn't PHB filling up our
supermarket shelves?
 A major factor limiting PHB's use is its brittleness. But is
used in the US navy (cups) and Japan (disposable
razors!)

GMO plants producing plastic?

 A team at the DoE Plant research lab at Michigan State
University took two genes from PHB-making bacteria
and inserted them cress plants. They had managed to
create a transgenic plant that could grow plastic.
 But scientists really are not yet sure about how easy it is
to confine genetically modified plants in one area, or
about the impact so much PHB would have on the
environment. It's too early yet to risk growing PHB on a
large scale outdoors…?