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toxicological manifestations of heavy metals along with the detailed description of bioremediation
technologies employed such as phytoremediation and biosorption for the potential removal of these
DOI: 10.1039/c3em00491k
metals. It also updates readers about recent advances in bioremediation technologies like the use of
rsc.li/process-impacts
Environmental impact
Levels of heavy metals are rising in the environment due to increased industrial usage causing severe damage to all spheres of life. Commonly followed methods
like ion exchange, chemical precipitation, reverse osmosis, bio-piles, bio-slurries and land-lling are not only expensive but their byproducts are hazardous to
the environment. Bioremediation is an upcoming technique which utilizes eco-friendly agents like enzymes, microorganisms and plants and can prove to be a
suitable alternative for the elimination of these heavy metals. It is imperative to carry out conclusive research which can rene and improve this process to a level
where it can be accepted universally. On this note this review throws light on the technology of bioremediation and discusses recent additions to this area.
Introduction
Pollution refers to the state of existence of undesirable
substances (pollutants) in the environment beyond a permissible limit which can harmfully aect every sphere of life.
Sources of pollution can be both natural and anthropogenic.
Natural sources include geothermal activities, comets, space
dust and volcanic activities. Whereas, anthropogenic sources
have arisen mainly on account of rapid industrialization and
extensive use of chemical substances such as hydrocarbons,
pesticides, chlorinated hydrocarbons and heavy metals.1 The
latter mentioned source is the major contributor to pollution in
contrast to the former.2 Out of a large number of aforementioned anthropogenic sources, toxicological manifestations
caused by heavy metals are well known and are considered as
highly detrimental.
Lead (Pb), cadmium (Cd), arsenic (As) and mercury (Hg) are
the major pollutants that bring about heavy metal toxicity. The
non-biodegradable nature of these metals is the principle
School of Biotechnology, Rajiv Gandhi Proudyogiki Vishwavidyalaya, Bhopal, M.P.,
India. E-mail: gaurnisha2007@gmail.com; Tel: +91 8234884887
reason that leads to their prolonged presence in the environment. Moreover, these metals can enter into the food chain and
over a period of time become accumulated in the human body.
This accumulation can cause many health eects which might
be irreversible in nature.3
Chelation therapy is the mainstay of the treatment regime
followed so far for curing heavy metal poisoning. However, this
therapy is coupled with severe side eects as apart from the
removal of toxic metals it also eliminates important minerals
and metals from the body like iron (Fe), calcium (Ca), zinc (Zn)
etc. which directly aects normal biological processes of the
body.4 Thus, rather than a curative approach using chelation to
treat heavy metal poisoning, a preventive approach can be an
eective alternative focusing on the eradication of these heavy
metals from the environment itself.
Conventional methods like ion exchange, chemical precipitation, reverse osmosis, bio-piles, bio-slurries, and land-lling
are used conventionally for the remediation of heavy metals
present in water and soil.5 However, they suer from a major
drawback of being expensive owing to the requirement of
sophisticated infrastructure. Moreover, they also generate toxic
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Mechanism of phytoremediation
Specically, the process of phytoremediation is broadly divided
into two phases for the sequestration of heavy metals from soil
and water: ex situ and in situ. The ex situ bioremediation process
for soil and water is a two-step method. Firstly, it involves the
excavation of contaminated soil or pumping out the groundwater for treatment. The soil and water is then subjected to
several chemical and physical methods like chemical reduction/
oxidation, dehalogenation, soil washing, uid vapour extraction, stabilization/solidication, and solvent extraction to
eradicate the contaminants.43 Thereaer, the treated soil or
water which is free from heavy metals is restored back to the
original site. The removed pollutants are then transported to
some other site for dumping.42 Although, this approach is less
time consuming and can be performed under controlled
conditions, due to dumping and o-site burial of the removed
contaminants at the time of treatment, it can act as another
threat to the environment at another location.44
Fig. 2
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Fig. 3
Mechanisms of phytoremediation.
Phytoextraction
Also known as phytoaccumulation, this process includes the
extraction of toxic metals from soil and water without disturbing its integrity. The absorption and uptake of heavy metals is
performed by plant roots followed by their translocation and
nally accumulation and concentration above ground in the
biomass (shoots).43 Generally, this method is favored for the
sites that are discreetly or supercially polluted.
The underlying mechanism behind phytoextration is
hyperaccumulation. It is the process in which the plants like
Pteris vittata L, Thlaspi rotundifolium (L.) Gaudin, Fagopyrum
esculentum Moench, and Betula papyrifera Marsh can accumulate toxic metals at relatively higher concentrations. Hyperaccumulation is widely favored on metalliferous soils (soils
aected with high concentrations of transition metals) and the
plants which grow on this type of soil are referred to as metallophytes.45 The plants used for phytoextraction should have
high hyper-accumulating capacity and should be capable of
growing on highly toxic soils (and water) in order to make the
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Phytoltration
Phytostabilization
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Phytovolatilization
This process uses plants for the uptake of contaminants from
soil and water followed by subsequent degradation into less
toxic forms which are then transpired into the environment.79
Plants can volatilize both organic and inorganic contaminants
provided that the inorganic contaminants should not form
methyl and hydride derivatives.80 Contaminants which have
high Henry's constant (KH is characteristic of particular solute,
solvent and temperature)81 i.e. KH> 10 atm-m3 water per (m3 air)
are applicable for the phytovolatilization mechanism.82
The mechanism includes open stomata of the leaves to
diuse volatile contaminants in the environment in less toxic
forms. The plants used in this process shows high levels of ux
of the pollutant towards the atmosphere through the transpiration process.80 This method not only removes the pollutants
from contaminated site in a volatile form but the removal is
done in safer forms of that particular pollutant.
Sakakibara et al. reported the eradication of As through a
remediation process by using Pterisvittata. They found that
Pterisvettata had a good eciency of volatilizing As (90%) from
arsenic-polluted soil. However, secondary arsenic pollution was
said to be caused if a large amount of arsenic is released into the
environment.83 In another study, the eect of ethylene glycol on
the phytovolatilization of 1,4-dioxane was estimated. DN34
poplar trees were used for this study and it was seen that when
10 g l 1 of ethylene glycol was present in ground water it
reduced the growth rate of plants to 28%. Similarly the eect of
ethylene glycol on Arabidopsis was also observed and it was
understood that ethylene glycol had an inhibitory eect on its
growth and under hydroponic conditions it inhibited the phytovolatilization of 1,4-dioxane.84 Carvalho et al. carried out
studies on four aquatic plants (Typha domingensis, Lemna
obscura, Hydrilla verticillata Royle and Crinum americanum) for
the removal of aqueous selenium. Initially, they found that the
plants accumulated the selenium in their tissues. But, later it
was concluded that the main mechanism behind selenium
accumulation was phytovolatilization. In this process, plants
converted the inorganic form of the selenium into the organic
form which is less toxic and was then transpired.85
The advantage of this technology is that it does not require
disposal of any contaminant thereby circumventing any site
disturbance and erosion. This process is restricted only for
abolition of volatile compounds and cannot be applied for the
removal of nonvolatile heavy metals. However, the main
disadvantage of phytovolatilization is that the heavy metals are
still toxic to some level even when they are volatilized. The rate
of their migration and translocation cannot be predicted in the
polluted area.62
Phytodegradation
This process exploits the capability of plants that possess
certain specialized enzymes (dehalogenase, reductase and oxygenase) or cofactors for the degradation of contaminants from
soil and groundwater.86 This method is limited only to organic
pollutants because these are biodegradable in nature. Phytodegradation diers from rhizodegradation mainly because of
the fact that the former encompasses the breakdown of
contaminants with the help of microorganisms present in the
rhizosphere and is a relatively slower mechanism. Flavonoids
and carbohydrates secreted by plants facilitating phytodegradation further enhance the microbial activity. Properties like
solubility, polarity, hydrophobicity and partitioning coecient
(Kd) of organic contaminants directly interferes with their entry
into plant through the root membrane.87 For the removal of
heavy metals some genetically modied plants have been
developed such as transgenic poplars.88
Farias et al. worked on petroleum-contaminated soil and
studied the tolerance and phytodegradation potential of Erythrina crista-galli L. in three dierent conditions: non-contaminated soil, vegetation-contaminated soil and non-vegetation
contaminated soil. They found that the growth of Erythrina
crista-galli L. in vegetation-contaminated soil was reduced as
compared to non-contaminated soil. On the other hand the
degradation of petroleum in vegetation-contaminated soil was
higher as compared to non-contaminated soil.89 Recently, a
study has been done on a transgenic tobacco plant which
expresses bacterial organophosphorus hydrolase (an enzyme
that degrades organophosphorus pesticides). Aer 14 days of
growth it was found that the tobacco plant degraded more than
92% of methyl parathion and gives more root and shoot
Biosorption
Biosorption is a process which uses biological materials for the
removal of contaminants through dierent mechanisms like
adsorption, absorption, surface complexation, precipitation
and ion exchange. It depends on numerous factors like
substance to be sorbed, environmental issues, biosorbent used,
presence and absence of metabolic process (in living organisms).91 The two terms absorption (process in which one
substance gets incorporated into another of dierent state) and
adsorption (physical phenomenon in which adherence and
binding of ions or molecule occur on the surface of another
molecule) comes under sorption process. In the case of
adsorption, the adsorbate is the substance which gets adsorbed
on a solid surface and the adsorbent is the soil surface.92 If the
adsorption phenomenon results in the formation of a stable
molecular phase at the interface, it is described as a surface
complex which can be of two types: inner and outer sphere
surface complexes. In the former one, the adsorbent gets bound
to at least one molecule of the hydration sphere of the adsorbate
but in the latter one without any hydration sphere the molecule
gets directly bound to the adsorbent.93,94
The contaminants that can be removed by biosorption could
be organic and inorganic or soluble and insoluble. Metals (K+,
Mg+) that are highly mobile and accordingly do not get accumulated with biomass during phytoremediation can be easily
removed through biosorption.95 Heavy metals (lead, arsenic,
cadmium, uranium, mercury) along with dyes, phenolic
compounds and pesticides are receiving a lot of attention for
their eradication through this process.96
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Types of biosorbent
Primarily biosorbents fall into the following categories: living
biomass and non-living biomass. Living biomass includes
bacteria (gram-positive bacteria, gram-negative bacteria and
cyanobacteria), fungi (mould, mushroom and yeast), algae
(micro-algae, macro-algae, brown seaweeds and red seaweeds).
While non-living biomass includes industrial waste (fermentation wastes, food/beverages waste, activated sludges, anaerobic
sludges), agricultural waste (fruit/vegetable waste, rice straws,
wheat bran, soybean hull etc.), natural residues (plant residues,
sawdust, tree bark, weeds etc.) and other biomaterials (chitosanbased materials, cellulose-based materials etc.).91,97
Many studies show that non-living biomass has gained
more preference over living biomass for the biosorption
process because it does not require any maintenance and
nutrient supply.98 Moreover, the biomass can be easily
obtained from industrial waste which adds the ease of availability and makes the process economic.99 Whereas, living
biomass demands proper maintenance of healthy microbial
culture coupled with sustained environmental conditions.
Even by providing these conditions, recovery of heavy metals
Fig. 4
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Biosorption mechanism(s)
The mechanism of biosorption is a highly complex process
owing to the complexity of the biological structures involved.
Functional groups like carboxyl, phosphate, hydroxyl, amino,
thiol etc. are present on the structure of biomass which interacts
with dierent heavy metals with variable degree which may be
aected by physico-chemical factors. There are number of
factors on which the binding of sorbate and sorbent depends
like number of binding site in the biosorbent, binding strength
of pollutant and functional groups present on the biosorbent,
availability and accessibility of sites.
There are various criteria on the basis of which mechanisms
of biosorption can be divided. This includes cell metabolism
Recent developments in
bioremediation
Recently new strategies for the process of bioremediation have
been uncovered. Scientists have shown the application of
nanoparticles, non-living biomass and genetically modied
plants for the removal of heavy metal toxicants from dierent
sources. These approaches are credited with having quick and
high bioremediation capacity.
Use of nanoparticles
Application of nanotechnology is widely being used for the
development of resourceful, ecient and environment friendly
nanomaterial systems in dierent spheres of biotechnology
including bioremediation. The physiochemical properties of
the nanoscale particles vary signicantly from their larger
counterparts. This is due to the very high surface to volume ratio
of the nanomaterials which provides them with high adsorption
capacity. Moreover, they have low cost and augmented
bioavailability which makes them excellent candidates for
bioremediation.117
Lately, superparamagnetic iron oxide nanoparticles (SPION)
have been used for the separation of contaminants from
wastewater because of their ultrane structure and high
competence. In this technique, the carriers contain a polymeric
shell having functional groups and a magnetic core (FeO, Fe3O4
and Fe2O3) which provides a strong magnetic response.118 Shen
et al., prepared and implemented Fe3O4 nanoparticles for the
purication of wastewater contaminated with heavy metals
(Cd2+, Cr6+, Cu2+ and Ni2+). The nanoparticles prepared were of
dierent size and were prepared by co-precipitation and polyol
method. They found that particles of 8 nm size were very
eective for the recovery and removal of metals from
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Conclusion
Levels of heavy metals are increasing day by day due to
increased industrial usage causing their accumulation in living
beings. Their exposure can cause fatal consequences to organ
systems through several mechanisms (primarily due to generation of oxidative stress). Oxidative stress leads to the production of free radicals followed by the decrease in the level of
antioxidants and nally leading to cell death.
Presently, conventional remediation methods like ion
exchange, chemical precipitation, reverse osmosis, landlling
and bio-piles are used for the removal of heavy metal contaminants. Although they have several advantages like ease of metal
Competing interest
The authors declare that they have no conict of interest.
Abbreviations
ROS
BBB
EDTA
Acknowledgements
I heartily acknowledge Ms. Batul Diwan (School of Biotechnology, Rajiv Gandhi Proudyogiki Vishwavidyalaya, Bhopal,
India) who played a major role in correcting and editing of the
manuscript.
References
1 V. Marcano, P. Benitez and E. Palacios-Pr
u, Planet. Space
Sci., 2003, 51, 159166.
2 I. G. Dubus, J. M. Hollis and C. D. Brown, Environ. Pollut.,
2000, 110, 331344.
3 R. Singh, N. Gautam, A. Mishra and R. Gupta, Indian J.
Pharmacol., 2011, 43, 246253.
4 S. J. Flora, M. Mittal and A. Mehta, Indian J. Med. Res., 2008,
128, 501523.
5 V. Vinod and R. Sashidhar, Indian J. Biotechnol., 2011, 10,
113120.
6 N. Singh and R. Gadi, J. Environ. Chem. Ecotoxicol., 2012, 4,
137142.
7 S. N. Singh and R. R. D. Tripathi, Environmental
bioremediation
technologies,
Springer-Verlag,
Berlin
Heidelberg, 2007.
8 A. N. Glazer and H. Nikaido, Microbial Biotechnology:
Fundamentals of Applied Microbiology, W.H. Freeman, 1995.
9 D. Di Baccio, R. Tognetti, L. Sebastiani and C. Vitagliano,
New Phytol., 2003, 159, 443452.
10 J. Kelly, M. H
aggblom and R. Tate, III, Biol. Fertil. Soils,
2003, 38, 6571.
11 R. A. Wuana and F. E. Okieimen, ISRN Ecol., 2011, 402647.
12 J. Buekers, Ph.D. thesis, Katholieke Universiteit Lueven,
Dissertationes De Agricultura, Doctoraatsprooefschri nr,
2007.
13 J. Shiowatana, R. G. McLaren, N. Chanmekha and
A. Samphao, J. Environ. Qual., 2001, 30, 19401949.
14 K. Jomova, D. Vondrakova, M. Lawson and M. Valko, Mol.
Cell. Biochem., 2010, 345, 91104.
15 K. A. Al-Ghanim, Afr. J. Biotechnol., 2011, 10, 1386013866.
16 R. R. Schroeder, Thesis for the Master of Environmental
Study Degree, The Evergreen State College, 2010.
17 G. Flora, D. Gupta and A. Tiwari, Interdiscip. Toxicol., 2012,
5, 4758.
18 S. Mahjoub and A. H. Moghaddam, Iranian Journal of Health
and Physical Activity, 2011, 2, 15.
19 J. Kasten-Jolly, Y. Heo and D. A. Lawrence, Toxicol. Appl.
Pharmacol., 2010, 247, 105115.
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