Environmental

Science

Processes & Impacts

View Article Online

CRITICAL REVIEW

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Cite this: DOI: 10.1039/c3em00491k

View Journal

A review with recent advancements on

bioremediation-based abolition of heavy metals
Nisha Gaur,* Gagan Flora, Mahavir Yadav and Archana Tiwari
There has been a signicant rise in the levels of heavy metals (Pb, As, Hg and Cd) due to their increased
industrial usage causing a severe concern to public health. The accumulation of heavy metals generates
oxidative stress in the body causing fatal eects to important biological processes leading to cell death.
Therefore, there is an imperative need to explore ecient and eective methods for the eradication of
these heavy metals as against the conventionally used uneconomical and time consuming strategies that
have numerous environmental hazards. One such eco-friendly, low cost and ecient alternative to
target heavy metals is bioremediation technology that utilizes various microorganisms, green plants or
enzymes for the abolition of heavy metals from polluted sites. This review comprehensively discusses

Received 26th September 2013

Accepted 12th November 2013

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

nanoparticles, non-living biomass and transgenic crops.

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

This journal is The Royal Society of Chemistry 2014

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

Environ. Sci.: Processes Impacts

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Environmental Science: Processes & Impacts

sludge which aects environment and might not completely

remove the metals.6
Bioremediation is one such eco-friendly and sustainable
process that can prove to be much more eective and ecient
for eliminating heavy metals present in dierent spheres of the
environment. This strategy aims to clean up the environment
while maintaining the normal biological processes associated
with it.7 According to Glazer and Nikaido, bioremediation is
dened as a process that uses microorganisms, green plants or
enzymes to treat the polluted sites for regaining their healthy
condition.8 This technique is highly favored as it provides much
better results through the application of low cost and economic
inputs in comparison to conventional means. Bioremediation
can thus be regarded as a highly cost-eective, eco-friendly and
more pronounced solution to the problems arising due to the
use of transition metals.

Heavy metal pollution in soil and water

Heavy metals such as Pb, As, Cd and Hg are ubiquitous in
nature and cause an unfavorable result on the surroundings
particularly at high concentrations. Even though the heavy
metals biochemical equivalence and geochemical cycles are
normal components of the earth's crust, their concentration has
become remarkably exacerbated following the advent of the
industrial revolution which resulted in a manifold rise in the
level of usage of these metals.9

Environ. Sci.: Processes Impacts

Critical Review

These heavy metals are known to facilitate phytotoxicity

through contamination of soil, a problem that has called for
considerable attention over the past few decades. The presence of heavy metals in soil can considerably decrease the size
of the microbial community along with reduction of environmental and biological activities such as organic matter
mineralization and leaf litter decomposition.10 The level of
contamination however depends on factors such as chemical
composition, toxicity, mobility and varying bioavailability of
the metal.11 As soon as these heavy metals come in contact
with the soil surface they initially become readily adsorbed,
which is followed by slow adsorption and distribution in
the soil.12,13 When the plants grow on metal-polluted soil they
tend to accumulate these heavy metals, which greatly aect
their growth and development. This not only threatens
their survival but also aects the life which consumes them.
Owing to this contamination, a large amount of terrain has
turned out to be dangerous and non-arable for humans and
animals.
In a manner similar to soil, both surface water and ground
water can easily become contaminated by heavy metals
through natural sources (leaching of ore, erosion of minerals
with sediments and volcanic extruded products) or human
activities (chemical fertilizers, pesticides, solid waste disposal,
industrial and domestic wastes). Due to its polarity and
hydrogen bonds, it may adsorb, dissolve and absorb many
dierent compounds.

Nisha Gaur received her M.Tech

in Biotechnology from Rajiv
Gandhi Technological University, Bhopal, India. She is
currently a research trainee at
R&D department of Kilpest India
Private Limited. Her research
interests include bioremediation
of organic and inorganic wastes.

Mahavir Yadav received his

Ph.D in Molecular Biology at the
Institute of Microbial Technology, Chandigarh, India. He is
currently working as an Assistant Professor in the School of
Biotechnology at Rajiv Gandhi
Technological University, Bhopal, India. His research interests
include biodiesel production
and bioremediation of heavy
metals.

Gagan Flora received his M.Tech

in Biotechnology from Rajiv
Gandhi Technological University, Bhopal, India. He will very
soon be starting his doctorate
studies. His research interests
include toxicity of heavy metals.

Archana Tiwari received her

Ph.D in Environmental Sciences
from Barkatullah University,
Bhopal, India. She is currently
working as an Associate
Professor and Head of the
School of Biotechnology, Rajiv
Gandhi Technological University, Bhopal, India. Her research
interests include production of
bioplastics from biological
sources.

This journal is The Royal Society of Chemistry 2014

View Article Online

Critical Review

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Toxicity of heavy metals

Metals having high atomic weight and a density of more than
5 g cm 3 are regarded as heavy metals or transition metals.
More than 20 dierent kinds of heavy metals are found in
nature but only a few of them are of concern to human health.
According to the Agency for Toxic Substances and Disease
Registry (ATSDR), lead, cadmium, arsenic and mercury are well
known for showing a toxicity prole upon exposure. Although
numerous cellular, intracellular and molecular mechanisms
have been reported to underpin heavy metal toxicity, generation
of oxidative stress is the well accepted mechanism which
explains most of the arising symptoms14 (Fig. 1). Chiey, human
exposure to these metals occurs from industries and toxic waste
sites. The non-biodegradable nature of these heavy metals
further results in their prolonged persistence in the environment. It is now known that existence of transition metals even at
a very low concentration (picomolar) in humans can result in
fatal health eects.15
1. Lead. Lead is a widely known ubiquitously present
xenobiotic heavy metal. Its unique properties like high ductility,
highly malleability, low melting point and soness makes it an
important metal in industries such as automobiles, paint,
ceramics, plastics etc.16 Due to this widespread usage, humans
have become vulnerable targets for its exposure. No level of lead
has been considered to be safe or benecial to living beings.
Upon exposure it aects many organs like the nervous system,
renal system, hematopoietic system, reproductive system and
cardiovascular system, and shows some eects on bone. The
nervous system is the most sensitive target compared to the
others for lead-induced toxicity.17,18
High exposure of lead may cause fatal consequences like
convulsions, lack of coordination, delirium and paralysis. It
also aects the hematopoietic system which inhibits the

Fig. 1 Generation of oxidative stress in cell owing to exposure of

heavy metals such as Pb+2, As+2, Cd+2, and Hg+2, leads to formation of
reactive oxygen species and impairs anti-oxidant defense causing cell
death.

This journal is The Royal Society of Chemistry 2014

Environmental Science: Processes & Impacts

synthesis of hemoglobin and thus causes anemia. Renal

dysfunction has also been reported on account of lead-induced
toxicity.19,20
The major mechanism of lead-induced toxicity is induction
of oxidative stress which occurs as a result of imbalance
between pro-oxidant and anti-oxidant ratio. This imbalance
brings about protein oxidation, lipid peroxidation and nucleic
acid peroxidation making a cell prone to cell death.21 The ionic
mechanism is the other mode of action of lead toxicity. In this
process, lead mimics and substitutes other monovalent and
bivalent ions like Na+, Ca2+, and Mg2+, and hinders many biological process like intracellular signaling, cell adhesion,
protein folding, ionic transportation etc.17
Chelation therapy has been regarded as the mainstay treatment which involves introduction of chelating agents like
calcium disodium ethylenediaminetetraacetic acid (CaNa2EDTA), D-penicillamine (DPA), Dimercaprol (BAL) and Succimer into the organism. These chelating agents then bind to
the lead ions forming a complex known as a chelate which is
excreted out of the body mainly through urine. Many natural
anti-oxidants like vitamins (B, C and E), avonoids, and herbal
avonoids have also been used for curing lead-induced toxicity.4
2. Cadmium. Cadmium is an extremely toxic metal having
distinctive properties such as good lustre, high ductility,
malleability and soness that have led to its extensive usage in
diverse industries like NiCd batteries, coatings and plating,
and as stabilizers for plastics.22
It causes many adverse health eects by damaging kidney,
liver, bone and cardiac tissues. The kidneys and liver are the
chief targets for cadmium-induced toxicity. Nephropathy is the
most common renal abnormality that occurs owing to cadmium
exposure. Renal vitamin D metabolism is also aected when
cadmium accumulates in the kidney.23 This signicantly brings
about a calcium imbalance which leads to osteoporosis and
osteomalacia as well as increased excretion of calcium (medical
condition referred to as Itai-Itai).24 Cadmium is also regarded as
a potent human carcinogen that is associated with high risk of
renal and prostate cancer. It is also known to act as a robust
mutagen and can cause multi-locus deletions.
Cadmium weakens the antioxidant defense by severely
reducing the intracellular glutathione levels. It also inhibits the
activity of various antioxidant enzymes like superoxide dismutase and catalase along with generation of ROS.25 The
combinatorial eect of these processes renders cells into a state
of oxidative stress. The increased level of ROS causes damage to
DNA and inhibits DNA repair resulting in mutation.23
3. Arsenic. According to the ATSDR, arsenic is regarded as
the most common cause of acute heavy metal poisoning in
adults and children.26 Arsenic is a ductile metalloid which exists
in three allotropic forms: metallic grey, yellow and black
arsenic. It is broadly used to make insecticides, fungicides,
weed killer, antifouling agents and in preserving woods.
Clinical manifestation of arsenic is referred to as Arsenicosis
and is caused by the prolonged exposure of arsenic in humans.
Pigmentation (development of spotty rain drop patches over the
front of chest) and keratosis are the toxic aermaths on the
skin.27 Arsenic toxicity leads to many respiratory diseases like

Environ. Sci.: Processes Impacts

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Environmental Science: Processes & Impacts

reduced pulmonary function, lung cancer, chronic cough or

chronic bronchitis. Peripheral neuritis, black foot disease, liver
brosis and gastroenteritis is also caused due to the uptake of
arsenic-contaminated water.28
Unlike lead and cadmium, the molecular mechanism
underlying arsenic toxicity is multi factorial. It involves generation of oxidative stress, suppression of DNA repair, inhibition
of cell cycle check points and induction of apoptosis.29
Chelation therapy is considered as the most preferred
approach to control the toxic eects of arsenic. Numerous
avonoids, vitamins and herbal extracts have also been reported for curing and preventing arsenic-mediated cellular and
molecular damage.26,30
4. Mercury. Mercury is a naturally occurring metal that can
exist in inorganic, organic (ethyl-, methyl-, alkyl-, or phenylmercury) and vapor states, with the organic state considered
more hazardous in comparison to other forms. It has both
industrial (batteries, fossil fuel emission, paints, cosmetic
products etc.) as well as clinical applications (thermometers,
sphygmomanometer, barometers etc.).31 In the environment, its
exposure occurs through the erosion of mercury-containing
ores and in the form of gases dissipating from volcanic eruptions which are rich in mercury.
The route of absorption of all forms of mercury is dierent.
95100% of organic mercury (methylmercury) is absorbed in the
intestinal tract and almost 100% gets inhaled through vapor.
The absorption rate of elemental mercury is less as compared to
organic mercury and is found to be around 7585% occurring
mainly through inhalation. Inorganic mercury is absorbed at
much lower rates (715%) of ingested dose and 23% of dermal
dose.31,32
Two forms of mercury, i.e. organic and elemental, are lipophilic in nature and become distributed throughout the body.
Both forms can cross the blood brain barrier (BBB) and even the
placental barrier and nally accumulate in the brain and
kidneys. Whereas, inorganic mercury is not capable of crossing
the BBB or placental barrier. It is found in the brain neonates
and accumulates in the kidneys.31,33
Mercury is a powerful neurotoxin which primarily aects the
central nervous system.34 It may lead to lack of coordination of
movements, impairment of speech and hearing, and muscle
weakness. The lungs absorbs metallic mercury through the
breathing process which can lead to respiratory impairment. It
has diverse mechanisms through which it can cause biochemical damage to tissues and genes. Mercury induces toxicity by
forming free radicals and generating oxidative stress.35 It also
bind to thiol-containing enzymes and inhibits them.31 Methylmercury forms complexes with cysteine, a thiol-containing
compound, which helps in intracellular absorption.36

Critical Review

hydrocarbons, organic solvents and crude oil from soil and

water to improve its quality.37,38
Although every plant has the capability to remove contaminants, only a few selected or engineered plants are used extensively to remove contaminants eciently such as Clerodendrum
infortunatum, Croton bonplandianus, Pistia stratiotes, Thlaspi
caerluescens, Brassica junceae, Alysum lesbiacum, etc.39 Phytoremedial strategies applied in context to heavy metal elimination
take into consideration any of the following methods depending upon the nature of the contaminant:
 Complete removal of the accumulated heavy metals.
 Degradation or containment of heavy metals.
 Combination of these.
Compared to conventional strategies being followed (in situ
vitrication, soil incineration, excavation and landll, soil
washing, soil ushing and solidication) phytoremediation is
an aesthetically pleasing, ecient and eco-friendly process in
removing contaminants from low to moderate levels.40,41 It is
also an economical method which reduces the cost to less than
the half the price of the conventional methods. Moreover, it
requires low installation and maintenance costs. It also
provides an added advantage by not only cleaning polluted soil
but by also preventing soil erosion and metal leaching.42

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

Phytoremediation: a robust strategy for

the eradication of toxic heavy metals
Phytoremediation is as an emerging technology that involves
application of selected plants to degrade, assimilate, metabolize
or detoxify undesirable substances like heavy metals, pesticides,

Environ. Sci.: Processes Impacts

Fig. 2

Dierent mechanisms involved in phytoremediation.

This journal is The Royal Society of Chemistry 2014

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Critical Review

Fig. 3

Mechanisms of phytoremediation.

The in situ method on the other hand is a technology which

removes the heavy metals from contaminated soil, water and air
without performing excavation and transport of contaminants.
The treatment regimes are carried out at the same site that
precludes o-site burial of the removed pollutants and thus
prevents contamination of the clean soil.42 This method causes
less ecological disturbance and is also economically viable
which makes it a better alternative than ex situ technology. This
method is further divided into dierent categories to remove
toxic metals from soil and water: phytoextraction, phytoltration, phytostabilization, phytovolatilization, phytodegradation,
and rhizodegradation (Fig. 2 and 3).

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

This journal is The Royal Society of Chemistry 2014

Environmental Science: Processes & Impacts

extraction and translocation process of toxic metals in the

shoots eective.46
In this process, regular cropping of the hyperaccumulator
plant is required until the metal concentration reduces down to
the desired level at the concerned site. Aer that the contaminated plant biomass is either burned and converted into ash or
is used in various industries (e.g. wood, cardboard, etc.). This
process can also be put into commercial use such as phytomining which essentially involves extraction of biomass in the
form of bio-ore (extracts of saleable heavy metals obtained by
the plant biomass ash).47
The phenomenon of hyperaccumulation is of two types:
natural hyperaccumulation and chemically-enhanced hyperaccumulation. Natural hyperaccumulation utilizes specic
kinds of hyperaccumulators capable of absorbing the toxic
metals in the roots followed by its translocation in shoots.
Finally, storage of these translocated heavy metals occurs the in
aerial portion of the plant in a nontoxic form. These plants have
high tolerance capacity and are credited with the ability of high
translocation mobility of metals by secreting metal chelating
compounds (phytosiderophores) and organic acids.48 Chemically-enhanced hyperaccumulation is applied in case of some
metals like lead and gold that are immobile in soil and cannot
be absorbed readily. For this purpose some chemical inducers
like chelating agents (EDTA, NTA, malate etc.) or acidifying
agents are used which enhance their mobility in the soil by
increasing the bioavailability of the metals in soil which ultimately boosts their uptake.49
Kaur et al. reported that the chemically-enhanced phytoextraction showed better accumulation capacity as compared to
the natural phytoextraction process. They also found that
Brassica juncea arawali can act as an excellent chemicallyenhanced hyperaccumulator.50 A recent study showed the
uptake of heavy metals from municipal solid waste by chelateassisted Festuca arundinacea. It was also reported that the
nitrilotriacetic acid signicantly enhanced the metal accumulation capacity of Festuca arundinacea in contrast to its
absence.51 In a phytoextraction analysis the eect of inducers
like EDTA on texturally dierent soil was assessed. It was
revealed that the induction of EDTA considerably enhanced the
lead accumulation capacity of wheat shoots in loamy sand than
that of the sandy clay loamy soil.52 In a recent study, assessment
of natural plants in Turkish serpentine soil was carried out for
analysis of its Ni accumulation capacity. Scientists tried to
establish the possible relationship between amount of phytoavailable Ni in the soil and the Ni content of potential accumulator plants. It was found that susceptibility and Ni requirement
of a plant was species specic. They presented Isati spinnatiloba
as potential Ni hyperaccumulator species.53
Phytoextraction can be performed by three means i.e. phytoextraction by trees, by crops and by grasses. Each has its own
advantages and disadvantages. Phytoextraction by trees
produces high biomass but due to shedding of leaves on to
surface the metal again becomes transported into the soil.
Phytoextraction by grasses has high metal accumulation
capacity but has low biomass production as well as slow growth
rate. Phytoextraction by crops has both the above mentioned

Environ. Sci.: Processes Impacts

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Environmental Science: Processes & Impacts

Critical Review

advantages. However, they pose a threat due to the ingestion of

crops by herbivores and thus entry of metals into the food
chain.54
There are four major steps involved in metal hyperaccumulation in plants:55
1. Bio-activation of trace metals in the rhizosphere.
2. Root adsorption and compartmentation with the help of
transporters and chelators.
3. Metal uptake by shoots.
4. Distribution, detoxication, and sequestration of metal
ions.
Hyperaccumulators need to have the facility of metal
homeostasis while growing in an impure surrounding. The
Thlaspi family are hyperaccumulating plants among which
twenty three species hyperaccumulate nickel, ten species
hyperaccumulate zinc, three species (T. caerulescens, T. praecox
and T. goesingense) hyperaccumulate cadmium and one species
hyperaccumulates lead.56 T. caerulescens is regarded as one of
the nest and well-known hyperaccumulators.57 Interestingly,
this plant is able to grow in serpentine soils, which contain
elevated levels of heavy metals including Zn, Co, Pb, Cr, Cd and
Ni, being capable of up taking up to 30 000 and 1000 mg kg 1
Zn and Cd, respectively in their shoots, while its development
remains unaected.58
Moreover, due to the advancement in genetic engineering,
the genes which help in remediation of heavy metals have been
isolated and then inserted into the large biomass producing
non-accumulating plants.59

advantages over rhizoltration because of the fact that seeds

can grow independent of environmental conditions and absorb
higher amounts of heavy metal during initial phase of their
life cycle.66
Removal of lead from wastewater using Carexpendula was
achieved by Yadav et al. using the rhizoltration technique.
They carried out pot and simulation experiments and found
that Carexpendula accumulated a signicant amount of lead
especially in root biomass as compared to shoot.67 In another
study, rhizoltration of cadmium and lead was performed by
using four dierent macrophytes (Pistiastratiotes L., Salvinia
auriculata Aubl, Salvinia minima Baker and Azollaliculoides
Lam). It was seen that Pistiastratiotes L. had extensive bioaccumulation eciency of removing lead and cadmium. The
accumulation of these two heavy metals in the roots was 10-fold
higher than that of the leaves.68 Vesely et al. reported the eciency of organic acids to enhance the mobility of heavy metals
through rhizoltration. They studied the bioaccumulation
potential of Pistiastratiotes L. against the removal of Cd, Pb and
Zn. They found that the organic acid substantially increased the
mobility of all heavy metals. However, translocation of heavy
metals decreased in the plant in a time-dependent manner.69
Rhizoltration has an advantage of absorbing metals readily
but this method works only in water and not in soil. Moreover,
metals get accumulated in the plant biomass which must be
disposed of regularly to reduce the risk of contamination.70

Phytoltration

Phytostabilization is a process which involves absorption and

precipitation of contaminants like heavy metals by plants
through immobilization. The process aims at the stabilization
of the heavy metal at the contaminated site instead of its
removal. This prevents movement of these contaminants via
ground water and wind.71 The underlying mechanisms that
determine the phenomenon of phytostabilization are as
follows:72,73
(a) Phytostabilization in the root zone: In this the root gets
exudated (converting contaminants into less bioavailable form)
in the rhizosphere so as to immobilize the heavy metal in the
root zone itself.
(b) Phytostabilization of the root membrane: This step leads
to the binding of the heavy metals to the root surfaces which
prevents their entry inside the plant.
(c) Phytostabilization in the root cells: This step further
prevents the translocation of heavy metals by sequestering them
into the cell vacuole.
For eective phytostabilization, the plants should have rich
root (to absorb large quantity of water) and shoot systems but a
poor translocation mechanism so as to prevent entry of heavy
metals into the shoots. Dense coverings of shoots tend to
increase transpiration which prevents precipitation of heavy
metals into the groundwater. Moreover, upward ow can be
maintained by fast transpiration by plants which prevents
downward leaching.62
Cambrolle et al. investigated the capability of two Spartina
species in terms of phytostabilization and bioaccumulation of

This process can be carried out in both terrestrial and aquatic

environments, though mainly it is carried out to purify ground
water or other water bodies.60 This process enables the plant
roots to absorb or adsorb, concentrate and precipitate the heavy
metals from a polluted euent source (industrial discharge,
agricultural runo, acid-mine drainage etc.). If plants use roots
for remediation purposes then it is known as rhizoltration.
Such kind of plants shows rapid growth of roots and removal
time of toxic metals is minimal.61
This process involves absorption of heavy metals by plants
roots followed by accumulation and transportation in the stem
or leaves. The contaminants are removed through harvesting at
an appropriate time.62 Development of feeder layer fertilization
(suspending several layers of soil above a polluted stream of
water through which the plant obtains the nutrients and
simultaneously removes heavy metals) system led to a boost in
rhizofertilization technology. Extensive root network development can be carried out by regular application of concentrated
fertilizers to this feeder layer.63
A second generation technology which uses plant seedlings
for the removal of heavy metals from contaminated water is
known as blastoltration.64 The seedlings have the potential to
absorb or adsorb a high percentage of toxic heavy metals.
Special kind of seedling cultures are prepared through
economic means using seeds, water and along with appropriate
exposure to light and darkness.65 This process shows

Environ. Sci.: Processes Impacts

Phytostabilization

This journal is The Royal Society of Chemistry 2014

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Critical Review

heavy metals like Co, Cr and Ni in two marshes with dierent

levels of contamination. They reported that in all the site
samples, the concentration of these heavy metals in both
species were higher in below-ground tissues as compared to the
above-ground tissues. Both species of Spartina showed good
phytostabilization capacity towards Co in the contaminated
soil.74 Varun et al. reported the phytostabilization potential of
Typhalatifolia L. in industrial sludge. Their ndings showed
that Typhalatifolia L. had higher potential for immobilization of
Zn, Mn, Cr and As but had less phytostabilisation potential
towards Ni, Cd and Co.75 Recently, the phytostabilization
capacity of six dierent plant species was assessed for the
immobilization of the lead in mine tailing using eld and pot
experiments. It was found that A. mangium had the best ability
for phytostabilization towards lead mine tailing out of six plant
species. A. mangium stored a higher concentration of lead in the
roots which aerward could be used in the timber industry or
paper industry etc.76 In another study, a Sorghum species was
used by Soudek et al. for the immobilization of heavy metals
(Zn, Cd) in soil deposited due to industrial activities. They
found that initially the root had a higher concentration of heavy
metal, but as the concentration of zinc and cadmium in the
solution increased they were transferred into the shoots which
ultimately caused toxicity to leaves. The toxicity aected the Chl
a/b ratio in the leaves.77
This technology is highly cost-eective in nature and does
not require the disposal of soil and contaminants aer treatment. However, this technology is not feasible for all sites
(restricted only to water) and also requires containment of
contaminants for an indenite period as they remain inside the
soil for a long time.78

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

This journal is The Royal Society of Chemistry 2014

Environmental Science: Processes & Impacts

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

Environ. Sci.: Processes Impacts

View Article Online

Environmental Science: Processes & Impacts

biomass as compared to the wild tobacco plant. This research

holds importance for the removal of organophosphorus
compounds from the environment.90

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

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

Critical Review

cannot be done eciently from living biomass as metals bind

intracellularly.98,100
Peptidoglycan carboxyl groups and phosphate groups
provide the metal binding sites in gram-positive and gramnegative bacteria respectively.101 In addition, the proteinaceous
S-layer and sheath also contribute to metal binding which are
made up of proteins and polysaccharides. The metal binding
component of many cyanobacterial cell walls contains peptidoglycan and some of them contain a sheath as well as extracellular polymeric substances.102 Pseudomurein is another cell
wall component which resembles peptidoglycans present in
archaea bacteria along with sulfonated polysaccharides and
glycoproteins which provides the anionic sites (carboxyl and
sulphate groups).103 Amongst all the components present in the
algal cell wall, cellulose is common in all algal diversity which
along with other components (depending on the presence) like
polysaccharides (mannan, alginic acid, xylans) and proteins
provides the binding sites (phosphate, sulphate, hydroxyl,
amine groups) for metal attachment.104
Chitins, glucans, mannans and proteins are the components
of fungal cell walls. Apart from these it also contains other
polysaccharides, lipids and pigments (melanin) which facilitate
binding of many metal ions.105 An important structural
component of the fungal cell wall is chitin which is cheaper as
compared to activated carbon and acts as an ecient biosorbent for metals as well as radionuclides.106 Like chitin, chitosan (derived from deacetylation of chitin) and other
derivatives of chitin also have eective biosorption capacity.107
Carboxyl, phenolic, hydroxyl, carbonyl and methoxyl groups are

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

Environ. Sci.: Processes Impacts

Mechanisms of biosorption can be categorised on the basis of

cellular metabolism and location of biosorption. Further, cellular
metabolism based biosorption is divided into metabolism-dependent
and non-metabolism-dependent. Metabolism-dependent includes
transport through cell membrane and precipitation. Non-metabolismdependent includes ion exchange, precipitation, complexation and
surface adsorption. On the basis of location, biosorption is classied as
extra cellular accumulation/precipitation, cell surface sorption/
precipitation, intracellular accumulation and further they are divided in
the same fashion as metabolism-dependent and non-metabolismdependent processes.

Fig. 4

This journal is The Royal Society of Chemistry 2014

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Critical Review

important as they bind to the oxygen binding sites which are

present in the phenolic polymers and melanins of the fungal
cell wall.93 Due to the availability of fungal biomass and rapid
growth rate these are receiving considerable attention as biosorbents among living cells.
As discussed above, due to the abundance and lesser processing requirement, agricultural and industrial waste and their
by-products are very economic and have received good acceptance. Adsorbents include leafs, bers, fruit peels, saw dust bark
etc. from agricultural and forest industries has been used in
removing metals from contaminated water. Due to their physico-chemical characteristics and their availability they can be
used as adsorbents.108
Vargas et al. worked on waste fruit cortex for the removal of
heavy metals from contaminated water. They checked the biosorption capacity of banana (Musa paradisiaca), lemon (Citrus
limonum) and orange (Citrus sinensis) peel. They found that
lemon and orange cortex showed good biosorption potential for
lead and copper as compared to banana. In the case of
cadmium, banana showed greater biosorption eciency than
lemon and orange. They also studied the relationship between
particle size and surface area and found them to be inversely
related to each other.109 In another investigation, Amaranthus
hybridus stalk and Carica papaya were used for removal of Mn
and Pb ions from wastewater. The study showed that among
both substrates Mn had greater percentage removal than lead.
The adsorptive capacity of Carica papaya in all cases was higher
as compared to Amaranthus hybridus stalk.110 In a recent study,
researchers worked on chitin and a-(1,3)-b-D-glucan (from
industrial bio-waste exhausted from brewer's yeast) for the
removal of heavy metals from acid mine drainage (Merladet and
Faith open-cast mines). They found that the Faith mine
drainage was contaminated with U, Al, Cu, Mn, etc. and the
Merladet mine drainage with Al, Mn, Zn, and Cu. They reported
that Saccharomyces cerevisiae L. acted as an ecient biosorbent
which eliminated heavy metals from polluted water.111 Suryan
et al. used paper mill waste for the removal of heavy metals (Pb,
Cd, Ni and Cu) from aqueous solution. They found that
adsorption process was aected by pH and adsorption rate in
case of all metal ions was above 70% (pH 2 to5). They concluded
that the paper mill waste did not require any pre-treatment and
recommended this as an option for better utilization of waste.112

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

This journal is The Royal Society of Chemistry 2014

Environmental Science: Processes & Impacts

and the location of biosorption (Fig. 4). Cell metabolism

mediated biosorption can further be divided into metabolismdependent and non-metabolism-dependent processes:113
 Metabolism dependent: occurs only in viable cells and
plays a vital role in the defence mechanism of microbes
showing reaction with toxic metals. Metabolism-dependent
biosorption may further be classied as:
Transport across cell membrane: microorganisms shares
the same mechanism for transport of heavy metals across the
membrane as well as transport of metabolic ions like sodium,
magnesium etc.88 It has no association with metabolic activity
and comprises two steps:
1. Metabolism-independent binding where the metals bind
to the cell walls.
2. Metabolism-dependent intracellular uptake which includes transport of metal ions across the cell membrane.
Precipitation: metabolism-dependent precipitation is
oen related to the microbe's defense mechanism where
their reaction in the presence of toxic metals produces
compounds favouring precipitation.114
 Non-metabolism-dependent: involves the interaction
between metal and functional groups present on the microbial
surface. As already discussed, many functional groups
(carboxyl, phosphate, sulphate etc.) are present on the microbial
surface because the microbial cell wall is made up of polysaccharides, proteins and lipids. It can further be categorised as
follows:
Physical adsorption: a physical phenomenon involving
van der Waal's and electrostatic forces. Even dead biomasses of
algae fungi and yeasts have shown adsorption of heavy metals
like copper, uranium, cobalt, zinc and cadmium through electrostatic interactions.93
Ion exchange: here, the polysaccharides of microbial cell
walls act as counter ions and facilitate the exchange of bivalent
metal ions. The marine algae alginates which occur as salts of
Mg2+, K+, Na+, Ca2+ can adsorb heavy metals by exchange of
counter ions like Co2+, Cu2+, Cd2+ and Zn2+.115
Complexation: interaction of metals with active groups
mediates their removal from solution via the formation of cell
surface complexes. Organic acids produced by microbes also
play an important role in chelating toxic metals by their solubilisation and leaching resulting in generation of metalloorganic molecules. Complexation and adsorption of metals is
brought about by carboxyl groups in microbial polysaccharides
and polymers.116
Precipitation: metabolism-independent precipitation
results from the metal and cell surface interaction which is a
chemical phenomenon.
The location-dependent biosorption can be categorised on
the basis of location where removed metals from solution
accumulate. It is of the following types:
 Extracellular accumulation/precipitation.
 Cell surface sorption/precipitation.
 Intracellular accumulation.
These can further be categorised in exactly the same fashion
as that in the case of metabolism-dependent and independent
biosorption.114

Environ. Sci.: Processes Impacts

View Article Online

Environmental Science: Processes & Impacts

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Merits and demerits of biosorption

Conventional methods like ion exchange, electrodialysis,
reverse osmosis etc. are expensive and their eciency of
removing heavy metals is very low. For this reason, biosorption
is receiving much attention because it shows many advantages
over traditional methods stated as follows:88
3 It is cost eective, selective and shows ecient removal of
metals even at low concentrations.
3 Unlike conventional methods it does not produce any toxic
sludge during the removal process which presents the opportunity of metal recovery and recycling of biosorbent.
3 Low cost adsorbents like industrial solid waste, agricultural waste etc. have shown excellent heavy metal removal
eciency.
3 It has a great advantage of being used in situ without the
need of any industrial process in integration with other ecofriendly systems.
3 With dead biomass, concerns related to toxicity, microbial
constrains and media formulation is alleviated.
The only disadvantage of using biosorbents is there is no
control over the biological characteristic of the biosorbent and
early saturation while performing experiments.

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

Environ. Sci.: Processes Impacts

Critical Review

wastewater. It was found that the adsorption capacity of Fe3O4

particles increased with decreasing the particle size or
increasing the surface area. Furthermore, maximum adsorption
was seen to occur at pH 4.0 under room temperature (20  C) and
the adsorption capacity of nanoparticles was found to be as
high as 35.46 mg g 1 (7 times higher than that of the coarse
particles).119 In another nding, the bioaccumulation capacity
of magnetic gel beads (prepared from Fe2O3 nanoparticle and
gellan gum) as potent bioadsorbents was assessed. They
concluded that the magnetic gel beads were eective for the
removal of lead, manganese and chromium in the order of
Pb2+ > Cr3+ > Mn2+. Additionally, it showed the high desorption
capacity with sodium citrate which would prove be very
economical.120 Bezbaruah et al., used calcium alginate beads for
the entrapment of zero-valent iron nanoparticle (Feo) to remove
the test contaminant present in ground water (nitrate). Zerovalent iron nanoparticles (nZVI) have been known to eradicate
various
groundwater
contaminants
like
chlorinated
compounds, pesticides and heavy metals. However, it suers
from higher mobility, agglomeration and settlement problems
by non-target compounds. They found that the overall removal
eciency of entrapped nZVI towards contaminants was
comparable to that of bare nZVI.121 Another investigation
reported the formulation of magnetic chitosan nanoparticles by
a one-step in situ co-precipitation method with an aim to
examine the sorption property for removing Cu(II) from aqueous
solution. The process was found to be highly competent and the
maximum sorption capacity was calculated to be 35.5 mg g 1.122

Use of non-living biomass

Another favorable strategy that is used extensively for bioremediation includes use of living microorganisms such as
bacteria, fungi, algae etc. They provide a large surface area to
volume ratio because of the smaller size of microorganisms.
However, they suer from several disadvantages such as
causing redox reactions between cell and medium which leads
to an increase in pH of the system. Also, this method increases
the biological oxygen demand and chemical oxygen demand as
it requires nutrient uptake.123 In contrast to this, the application
of dead biomass can be regarded as a suitable and favorable
alternative. Dead biomass does not require growth media or
nutrients and does not cause toxicity during the metal removal
process. Besides, this method is very economical in comparison
to living biomass.124
A recent investigation showed the biosorption capability of
nonliving biomass of marine macrophytes for arsenic removal.
Dierent phyla of alga were used to check their capability under
dierent pH conditions. Considerable adsorption was exhibited
by all the species. The highest observed value was by red alga
Ceramium (1.3  0.1 mg g 1) and seagrass Zostera, comparable
to other low-cost adsorbents like activated carbon. It was also
observed that there were many factors on which biosorption
rested like composition and structure of outer layer of macrophytes, availability of functional groups present on arsenic,
dierent pH levels and counter ionic interaction with arseniate.125 Ekmekyapar et al., studied the biosorption capacity of

This journal is The Royal Society of Chemistry 2014

View Article Online

Critical Review

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Cladonia rangiformis (a non-living lichen) for the removal of lead

from aqueous solution. They found that the non-living lichen is
a natural biomass, harmless and easily available and exhibited a
high accumulation capacity towards lead which can be exploited for the treatment of industrial euents.126 Mohamad et al.,
reported the capability of dead cells of Mesorhizobium amorphae
to act as a robust biosorbent for the removal of Cu2+ from
aqueous solutions.123 Biosorption capacity of non-living
biomass of Spirulina sp. for the removal of lead and zinc was
also reported in a work carried out by Goyal et al.127
Use of genetically modied plants (GMP)
The main aim of genetic engineering in the eld of phytoremediation is to enhance the capacity of plants to tolerate,
accumulate and absorb contaminants. Many genes from
dierent organisms have been identied and characterized that
are involved in acquisition, allocation and decontamination of
metals. The recombinant proteins produced by these transgenic
plants play an important role in chelation (e.g. citrate, phytochelatins, metallothioneins, phytosiderophores and ferritin),
assimilation and membrane transport of metals.128
Recently transgenic Arabidopsis thaliana was developed to
increase the tolerance and accumulation of arsenic and
cadmium by simultaneous over-expression of AsPCS1 and YCF1
genes. These genes are derived from garlic and baker's yeast.
This work was based on chelation of metals and vacuolar
compartmentalization which are the main strategies for heavy
metals/metalloids detoxication. It was found that the two
genes simultaneously increased the accumulation capacity as
compared to use of a single gene.129 Zhang et al., carried out a
study on a transgenic alfalfa plant with a motive to enhance its
resistance capability towards the heavy metals and organic
pollutants. This transgenic plant expressed both human
CYP2E1 and glutathione S-transferase which were produced
from hypocotyl segments by the use of Agrobacterium-mediated
transformation. They found that the transgenic alfalfa plant
which expressed both genes simultaneously had a remarkable
potential to remove mixed contaminants as compared to the
wild type and transgenic plant expressing the single gene.130
Another GMP, Nicotiana tabacum carrying a yeast metallothionein gene was shown to accumulate cadmium in the root
of the transgenic plant.131

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

This journal is The Royal Society of Chemistry 2014

Environmental Science: Processes & Impacts

recovery, pure euent production and high productivity, they

have severe disadvantages which includes their high cost,
production of toxic sludge and incomplete removal of metals.
So, as a potential alternative to these methods, bioremediation is a promising upcoming technology which uses plants,
microbes and their enzymes for the removal of heavy metals in
an eco-friendly manner. Bioremediation involves two
approaches i.e. phytoremediation and biosorption. Phytoremediation makes use of plants which have the capability to
accumulate, degrade and/or volatilize the heavy metals, hydrocarbons and organic solvents leading to improvement in the
quality of soil and water. Depending upon the conditions for e.g.
on the basis of the site (soil or water) and property of contaminants (organic or inorganic), phytoremediating plants implement dierent mechanisms which include phytoextraction,
phytoltration, phytostabilization, phytovolatilization and
phytodegradation.
Another mode of bioremediation exists called biosorption
which uses low cost adsorbents like industrial waste, agricultural waste, microbial biomass and their derivatives for the
treatment of aqueous waste. Dierent mechanisms are used by
the adsorbent on the basis of location of biosorption and
cellular metabolism. Further, they are divided into ion
exchange, precipitation, complexation and physical adsorption.
The heavy metal adsorption depends upon the type of adsorbent used, surface area, particle size, shape of the adsorbent
and experimental conditions. Biosorption is a promising
approach and is not only cost eective but also shows selectivity
and high eciency towards the removal of heavy metals.
Moreover, it does not produce any toxic sludge.
The latest addition to this technology is the application of
nanoparticles, non-living biomass and transgenic crops. These
novel approaches carrying out bioremediation have given highly
encouraging results and in addition of being ecient they are
also economical and give rapid results.
Thus, bioremediation has bright prospects for the abolition
of heavy metals from polluted sites. Also, its applicability could
be further enhanced by identifying and implementing more
novel plants and biosorbents that can oer better scope for
heavy metal removal without the need for any substrate modication. Consequently, the processing cost for modication of
adsorbents could be saved. Conclusively, it can be said that the
bioremediation technology has given us a platform that could
direct us towards the elimination of heavy metal pollution in an
eco-friendly manner.

Competing interest
The authors declare that they have no conict of interest.

Abbreviations
ROS
BBB
EDTA

Reactive Oxygen Species

Blood Brain Barrier
Ethylenediaminetetraacetic acid

Environ. Sci.: Processes Impacts

View Article Online

Environmental Science: Processes & Impacts

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

CaNa2EDTA Calcium disodium ethylenediaminetetraacetic

acid
DPA
D-Penicillamine
BAL
Dimercaprol
NTA
Nitrilotriacetic acid
Chl
Chlorophyll
KH
Henry constant
Kd
Partitioning coecient.

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.

Environ. Sci.: Processes Impacts

Critical Review

20 N. Fretellier, N. Bouzian, N. Parmentier, P. Bruneval,

G. Jestin, C. Factor, C. Mandet, F. Daubin
e, F. Massicot
and O. Lapr
evote, Toxicol. Sci., 2013, 131, 259270.
21 B. Anilkumar, A. G. Reddy, P. Ravikumar, Y. R. Reddy and
C. Haritha, Inventi Rapid: Molecular Pharmacology, 2012,
Inventi:pmp/167/12.
22 T. O. Llewellyn, Cadmium (materials Flow), Department of
the Interior. Bureau of Mines, United States, 1994.
23 V. Arroyo, K. Flores, L. Ortiz, L. G
omez-Quiroz and
M. Guti
errez-Ruiz, J. Drug Metab. Toxicol., 2012, 5, 2.
24 H. Baba, K. Tsuneyama, M. Yazaki, K. Nagata,
T. Minamisaka, T. Tsuda, K. Nomoto, S. Hayashi, S. Miwa
and T. Nakajima, Mod. Pathol., 2013, 12281234.
25 M. Filipic, T. Fatur and M. Vudrag, Hum. Exp. Toxicol., 2006,
25, 6777.
26 G. J. Flora, Curr. Trends Biotechnol.Pharm., 2012, 6, 280289.
27 S. Chanda, B. Ganguli, U. B. Dasgupta and D. G. Mazumder,
J. Toxicol. Environ. Health Sci., 2011, 3, 171175.
28 D. Guha Mazumder and U. Dasgupta, Kaohsiung J. Med. Sci.,
2011, 27, 360370.
29 K. Jomova, Z. Jenisova, M. Feszterova, S. Baros, J. Liska,
D. Hudecova, C. Rhodes and M. Valko, J. Appl. Toxicol.,
2011, 31, 95107.
30 V. Pachauri, A. Mehta, D. Mishra and S. J. Flora,
Neurotoxicology, 2013, 137145.
31 L. Patrick, Alternative Med. Rev., 2002, 7, 456471.
32 J.-D. Park and W. Zheng, Journal of Preventive Medicine and
Public Health, 2012, 45, 344352.
33 M. Korbas, J. L. O'Donoghue, G. E. Watson, I. J. Pickering,
S. P. Singh, G. J. Myers, T. W. Clarkson and G. N. George,
ACS Chem. Neurosci., 2010, 1, 810818.
34 C. Tschirhart, P. Handschumacher, D. Lay and
E. B
en
ece, Hum. Ecol., 2012, 40, 511523.
35 M. Valko, H. Morris and M. T. Cronin, Curr. Med. Chem.,
2005, 12, 11611208.
36 A. P. Society and H. Press, Cell Physiology, American
Physiological Society, 1992.
37 A. Erakhrumen, Educational Research and Review, 2007, 2,
151156.
38 N. Adams, D. Carroll, K. Madalinski, S. Rock, T. Wilson and
P. Pivetz, Introduction to phytoremediation, National Risk
Management Research Laboratory, Cicinnati, Ohio, USA,
2000.
39 P. Ahmadpour, F. Ahmadpour, T. Mahmud, A. Abdu,
M. Soleimani and F. H. Tayefeh, Afr. J. Biotechnol., 2012,
11, 1403614043.
40 Y. D. Jing, Z. L. He and X. E. Yang, J. Zhejiang Univ., Sci., B,
2007, 8, 192207.
41 M. I. Lone, Z. L. He, P. J. Stoella and X. E. Yang, J. Zhejiang
Univ., Sci., B, 2008, 9, 210220.
42 M. Ghosh and S. Singh, Applied Ecology and Environmental
Research, 2005, 3, 118.
43 S. Hayat and A. Ahmad, Brassinosteroids: A Class of Plant
Hormone, Springer London, Limited, 2011.
44 L. V. Pavel and M. Gavrilescu, Environ. Eng. Manage. J., 2008,
7, 815834.
45 N. Rascio and F. Navari-Izzo, Plant Sci., 2011, 180, 169181.

This journal is The Royal Society of Chemistry 2014

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Critical Review

46 M. M. Lasat, The use of plants for the removal of toxic metals

from contaminated soils, American Association for the
Advancement of Science, Environmental Science and
Engineering Fellow, Washington, D.C., USA, 2000, p. 33.
47 V. Sheoran, A. S. Sheoran and P. Poonia, Miner. Eng., 2009,
22, 10071019.
48 T. Mahmood, Soil Environ., 2010, 29, 91109.
49 B. V. Tangahu, S. Abdullah, S. Rozaimah, H. Basri, M. Idris,
N. Anuar and M. Mukhlisin, Int. J. Chem. Eng., 2011, 2011,
939161.
50 L. Kaur, K. Gadgil and S. Sharma, International Journal of
Bioassays, 2013, 2, 352357.
51 S. Zhao, L. Jia and L. Duo, Bioresour. Technol., 2013, 129,
249255.
52 Saifullah, M. H. Zia, E. Meers, A. Ghafoor, G. Murtaza,
M. Sabir, M. Zia-ur-Rehman and F. M. G. Tack,
Chemosphere, 2010, 79, 652658.
nver, Turk. J. Bot.,
53 H. Altin
ozl
u, A. Karag
oz, T. Polat and I. U
2012, 36, 269280.
54 W. A. Number, C. Ma, J. Kingscott and M. Evans, U.S.
Environmental Protection Agency Oce of Solid Waste and
Emergency Response Technology Innovation Oce,
Washington D.C., 1997.
55 H. Patra and M. Mohanty, Development, 2013, 25, 27.
56 S. Mukhopadhyay and S. Maiti, Global J. Environ. Res., 2010,
4, 135150.
57 C. Cosio, E. Martinoia and C. Keller, Plant Physiol., 2004,
134, 716725.
58 J. B. Wells, T. U. o. T. a. S. A. Earth and E. Science, Eect of
Infection with Glomus Mosseae on Cadmium Uptake in Vetiver
Grass [Chrysopogon Zizanioides (L.) Roberty], University of
Texas, San Antonio, 2008.
59 M. L. Lang, Y. X. Zhang and T. Y. Chai, Chin. J. Biotechnol.,
2004, 20, 157164.
60 M. S. Islam, Y. Ueno, M. T. Sikder and M. Kurasaki, Int. J.
Phytorem., 2012, 10101021.
61 D. Sarkar, R. Datta and R. Hannigan, Concepts and
Applications in Environmental Geochemistry, Elsevier
Science, 2011.
62 N. A. Anjum, M. E. Pereira, I. Ahmad, A. C. Duarte and
S. Umar, Phytotechnologies: Remediation of Environmental
Contaminants, CRC Press Inc, 2012.
63 S. K. Ong, Natural Processes and Systems for Hazardous
Waste Treatment, American Society of Civil Engineers,
2008.
64 P. D. M. Jumba, Genetically Modied Organisms The Mystery
Unraveled, AEG Literary Publishing Services, Incorporated,
2010.
65 I. Raskin, R. D. Smith and D. E. Salt, Curr. Opin. Biotechnol.,
1997, 8, 221226.
66 B. Thomas, D. J. Murphy and B. G. Murray, Encyclopedia of
Applied Plant Sciences, Three-Volume Set, Elsevier Science,
2003.
67 B. K. Yadav, M. A. Siebel and J. J. A. van Bruggen, Clean: Soil,
Air, Water, 2011, 39, 467474.
68 T. Vesely, P. Tlustos and J. Szakova, Int. J. Phytorem., 2011,
13, 859872.

This journal is The Royal Society of Chemistry 2014

Environmental Science: Processes & Impacts

69 T. Vesely, P. Tlustos and J. Szakova, Int. J. Phytorem., 2012,

14, 335349.
70 C. S. K. Mishra, Environmental Biotechnology, APH
Publishing Corporation, 2007.
71 R. Mohee and A. Mudhoo, Bioremediation and
Sustainability: Research and Applications, Wiley, 2012.
72 S. Shilev, M. Benlloch, R. Dios-Palomares and E. D. Sancho,
in Predictive Modeling and Risk Assessment, Springer, 2009,
pp. 225242.
73 R. Costa and K. Kristbergsson, Predictive Modeling and Risk
Assessment, Springer, USA, 2009.
74 J. Cambroll
e, E. Mateos-Naranjo, S. Redondo-G
omez,
T. Luque and M. Figueroa, Hydrobiologia, 2011, 671,
95103.
75 M. Varun, R. D'Souza, J. Pratas and M. Paul, Biotechnol.
Bioinf. Bioeng., 2011, 1, 137145.
76 W. Meeinkuirt, P. Pokethitiyook, M. Kruatrachue,
P. Tanhan and R. Chaiyarat, Int. J. Phytorem., 2012, 14,
925938.
77 P. Soudek,
S. Petrov
a and T. Van
ek, International
Proceedings of Chemical, Biological and Environmental
Engineering (IPCBEE), 2012, 46, 2529.
78 L. K. Wang, N. K. Shammas and Y. T. Hung, Waste
Treatment in the Metal Manufacturing, Forming, Coating,
and Finishing Industries, Taylor & Francis, 2010.
79 S. Feroz, D. G. Rao, R. Senthilkumar and J. A. Byrne,
Wastewater
Treatment:
Advanced
Processes
and
Technologies, Taylor & Francis Group, 2012.
80 B. P. Singh, Industrial Crops and Uses, CABI Publishing,
2010.
81 D. L. Reger, S. R. Goode and D. W. Ball, Chemistry, Principles
and Practice, BROOKS COLE Publishing Company, 2009.
82 P. J. Alvarez and W. A. Illman, Bioremediation and Natural
Attenuation: Process Fundamentals and Mathematical
Models, Wiley, 2005.
83 M. Sakakibara, A. Watanabe, M. Inoue, S. Sano and
T. Kaise, Proceedings of the Annual International Conference
on Soils, Sediments, Water and Energy, 2010.
84 M. R. Edwards, M. F. Hetu, M. Columbus, A. Silva and
D. D. Lefebvre, Int. J. Phytorem., 2011, 13, 702716.
85 K. M. Carvalho and D. F. Martin, J. Aquat. Plant Manage.,
2001, 39, 3336.
86 P. A. Kulakow and V. V. Pidlisnyuk, Application of
Phytotechnologies for Cleanup of Industrial, Agricultural and
Wastewater Contamination, Springer, 2009.
87 D. Tsao and M. K. Banks, Phytoremediation, Springer, 2003.
88 H. Fulekar, Bioremediation Technology: Recent Advances,
Springer, Netherlands, 2010.
89 V. de Farias, L. T. Maranho, E. C. de Vasconcelos, M. A. da
Silva Carvalho Filho, L. G. Lacerda, J. A. M. Azevedo,
A. Pandey and C. R. Soccol, Appl. Biochem. Biotechnol.,
2009, 157, 1022.
90 X. Wang, N. Wu, J. Guo, X. Chu, J. Tian, B. Yao and Y. Fan,
Biochem. Biophys. Res. Commun., 2008, 365, 453458.
91 G. M. Gadd, J. Chem. Technol. Biotechnol., 2009, 84, 1328.
92 W. Stumm and J. J. Morgan, Aquatic Chemistry: Chemical
Equilibria and Rates in Natural Waters, Wiley, 2012.

Environ. Sci.: Processes Impacts

View Article Online

Published on 12 November 2013. Downloaded by RSC Internal on 23/12/2013 10:52:49.

Environmental Science: Processes & Impacts

93 G. Crini and P. M. Badot, Sorption processes and pollution:

Conventional and non-conventional sorbents for pollutant
removal from wastemasters, Presses Universitaires de
Franche-Comt
e, 2011.
94 G. Sposito, The Chemistry of Soils, Oxford University Press,
USA, 2008.
95 Z. Aksu, Process Biochem., 2005, 40, 9971026.
96 G. M. Gadd, FEMS Microbiol. Lett., 1992, 100, 197203.
97 D. Park, Y.-S. Yun and J. Park, Biotechnol. Bioprocess Eng.,
2010, 15, 86102.
98 K. Vijayaraghavan and Y.-S. Yun, Biotechnol. Adv., 2008, 26,
266291.
99 R. H. Vieira and B. Volesky, Int. Microbiol., 2000, 3,
1724.
100 J. Modak and K. Natarajan, Miner. Metall. Process., 1995, 12,
189196.
101 P. Kotrba, M. Mackova and T. Macek, Microbial biosorption
of metals, Springer, Netherlands, 2011.
102 G. M. Gadd, Microbiology, 2010, 156, 609643.
103 G. Seltmann and O. Holst, The Bacterial Cell Wall, Springer,
2002.
104 T. A. Davis, B. Volesky and A. Mucci, Water Res., 2003, 37,
43114330.
105 J.-P. Latg
e, Cell. Microbiol., 2010, 12, 863872.
106 J. Wang and C. Chen, Biotechnol. Adv., 2009, 27, 195226.
107 E. Lichtfouse, J. Schwarzbauer and D. Robert,
Environmental Chemistry for a Sustainable World:
Remediation of air and water pollution, Springer
Science+Business Media B.V., 2011.
108 M. Rafatullah, O. Sulaiman, R. Hashim and A. Ahmad,
J. Hazard. Mater., 2010, 177, 7080.
109 K. Kelly-Vargas, M. Cerro-Lopez, S. Reyna-Tellez,
E. R. Bandala and J. L. Sanchez-Salas, Phys. Chem. Earth,
2012, 3739, 2629.
110 J. Egila, B. Dauda, Y. Iyaka and T. Jimoh, Int. J. Phys. Sci.,
2011, 6, 21522157.
111 F. Ramrez-Paredes, T. Manzano-Mu
noz, J. Garcia-Prieto,
G. Zhadan, V. Shnyrov, J. Kennedy and M. Roig,
Biotechnol. Bioprocess Eng., 2011, 16, 12621272.
112 S. Suryan and S. Ahluwalia, International Journal of
Environmental Sciences, 2010, 13311343.

Environ. Sci.: Processes Impacts

Critical Review

113 N. Ahalya, T. Ramachandra and R. Kanamadi, Res. J. Chem.

Environ., 2003, 7, 7179.
114 L. K. Wang and J. H. Tay, Environmental Bioengineering,
Humana, 2010.
115 S. K. Sharma and A. Mudhoo, Green Chemistry for
Environmental Sustainability, Taylor & Francis, 2010.
116 S. Srivastava and P. Goyal, Novel Biomaterials:
Decontamination of Toxic Metals from Wastewater,
Springer, Berlin Heidelberg, 2010.
117 S. Cameotra and S. Dhanjal, in Bioremediation Technology,
ed. M. H. Fulekar, Springer, Netherlands, 2010, pp. 348
374.
118 V. V. Mody, R. Siwale, A. Singh and H. R. Mody, J. Pharm.
BioAllied Sci., 2010, 2, 282.
119 Y. F. Shen, J. Tang, Z. H. Nie, Y. D. Wang, Y. Ren and L. Zuo,
Sep. Purif. Technol., 2009, 68, 312319.
120 X. Wang, C. Zhao, P. Zhao, P. Dou, Y. Ding and P. Xu,
Bioresour. Technol., 2009, 100, 23012304.
121 A. N. Bezbaruah, S. Krajangpan, B. J. Chisholm, E. Khan
and J. J. Elorza Bermudez, J. Hazard. Mater., 2009, 166,
13391343.
122 C. Yuwei and W. Jianlong, Chem. Eng. J., 2011, 168, 286
292.
123 O. A. Mohamad, X. Hao, P. Xie, S. Hatab, Y. Lin and G. Wei,
Microbes Environ., 2012, 27, 234241.
124 C. M. Monteiro, P. M. Castro and F. X. Malcata, Biotechnol.
Prog., 2012, 28, 299311.
125 C. Pennesi, F. Vegli`
o, C. Totti, T. Romagnoli and
F. Beolchini, J. Appl. Phycol., 2012, 24, 14951502.
126 F. Ekmekyapar, A. Aslan, Y. Bayhan and A. Cakici, Int. J.
Environ. Res., 2012, 6, 417424.
127 R. K. Aneja, G. Chaudhary, S. S. Ahluwalia and D. Goyal,
Indian J. Microbiol., 2010, 50, 438442.
128 T. Vamerali, M. Bandiera and G. Mosca, Environ. Chem.
Lett., 2010, 8, 117.
129 J. Guo, W. Xu and M. Ma, J. Hazard. Mater., 2012, 199200,
309313.
130 Y. Zhang and J. Liu, J. Hazard. Mater., 2011, 189, 357362.
131 O. Krystofova, O. Zitka, S. Krizkova, D. Hynek,
V. Shestivska, V. Adam, J. Hubalek, M. Mackova, T. Macek
and J. Zehnalek, Int. J. Electrochem. Sci., 2012, 7, 886907.

This journal is The Royal Society of Chemistry 2014