International Microbiology

https://doi.org/10.1007/s10123-018-0012-3

REVIEW

Adverse effect of heavy metals (As, Pb, Hg, and Cr) on health and their
bioremediation strategies: a review
Amit Pratush 1 & Ajay Kumar 2 & Zhong Hu 1

Received: 9 April 2018 / Revised: 23 May 2018 / Accepted: 28 May 2018

# Springer International Publishing AG, part of Springer Nature 2018

Abstract
Currently, heavy metal pollution becomes a severe problem whole over the world, and these toxic metals enter into the envi-
ronment either by natural phenomena or due to extensive industrialization. The discharged effluents containing toxic heavy
metals mixed with soil/water and change their natural composition. These heavy metals have adverse effects on living beings and
cause damage to the vital body organs of animals as well as humans. The heavy metal pollution also inhibits the biodegradation of
the chlorinated organic compounds (another type of environmental pollution) by interacting with metabolizing enzymes and
inhibits their functioning. Earlier studies described that heavy metals cannot be fully removed from the environment, but they can
be effectively neutralized or transformed into less toxic form so that their harmful effect on the environment can be reduced. The
distinctive enzymatic apparatus within microbial system plays a major role in the transformation of heavy metals in the envi-
ronment. A considerable advancement has been made during recent years to transform the heavy metals by utilizing the
bioremediation potential of genetically engineered (GE) microorganisms. These transgenics are very much efficient in heavy
metal transformations and still, we have to discover more to additionally utilize their full biotransformation potential.
In the present review article, the detailed description of the adverse effects of four heavy metals (arsenic, lead, mercury, and
chromium) and their adverse effect on our environment and human beings is discussed. Furthermore, the use of microorganisms/
GE organisms for the bioremediation of heavy metals from the environment is also discussed along with their detailed biore-
mediation mechanism.

Keywords Heavy metals . Biotransformation . Micro remediation . Contaminant . Pollution

Introduction
The fast rise of industrialization boosts the economy of the
country, but it also leads to the environmental pollution by
liberating pollutants such as toxic wastes of heavy metals like
arsenic (As), lead (Pb), mercury (Hg), and chromium (Cr),
smoke, and fumes. (Hansda et al., 2014). Presently, pollution
especially heavy metal pollution becomes one of the major
* Amit Pratush
pratush@stu.edu.cn
* Zhong Hu
hzh@stu.edu.cn
1
Biology Department, College of Science, Shantou University,
Shantou 515063, China
2
Department of Microbiology, School of Biological Sciences, Faculty
of Natural and Health Sciences, North-West University,
Potchefstroom 2735, South Africa
issues all over the world. The industries like mining, smelting,
and metal treatment lead to the discharge of a large amount of
heavy metals in the environment which is a cause of concern
for the public as well as to environmental health (Parmar &
Thakur, 2013).
The industrial wastes containing toxic heavy metal wastes
are mostly water soluble and are easily mixed with the soil or
water which changes the natural composition of these me-
diums (Ramamurthy & Memarian, 2012). Due to nondegrad-
able nature of these heavy metals, they enter the soil/water and
finally contaminate the food chain (Azimi et al., 2017). These
toxic wastes also affect soil fertility drastically as a result of
which both quality and quantity of food production, reduce
(Chibuike & Obiora, 2014). The most of the heavy metal
pollution is severe, long-term, and non-reversible in nature
(Tang et al., 2014). The most common heavy metals which
act as pollutants include arsenic (As), lead (Pb), mercury (Hg),
chromium (Cr), zinc (Zn), cadmium (Cd), copper (Cu), and
nickel (Ni) (Hansda et al., 2014). It has been observed that a

limited concentration of these heavy metals is necessary for
maintaining a proper metabolism in living beings, but a high
concentration of these heavy metals causes many serious
problems in both plants and animals (Govind, 2014). In hu-
man, the heavy metals like Hg, As, and Pb showed their toxic
effect on the kidney and nervous system which further leads to
mental disorders along with weakness, headache, abdominal
cramps, diarrhea, and anemia (Sharma et al., 2014). The de-
tailed description of all these eight heavy metals (As, Pb, Hg,
Cr, Zn, Cd, Cu, and Ni) including their contamination sources
(natural and anthropogenic), uses, and their adverse effect on
health is given in Table 1.
Currently, the number of methods is available like chemical
precipitation, dialysis, ion exchange, reverse osmosis, and sol-
vent extraction to remove these toxic heavy metals from the
contaminated system (Xu et al., 2017). Although, these tech-
niques are costly and have very less efficiency or sometimes
they also showed adverse effects on soil and change its orig-
inal composition (Azimi et al., 2017). To overcome these lim-
itations (low efficiency and adverse effects), other methods are
also invented which are eco-friendly and have no adverse
effects (Uqab et al., 2016). These methods are known as bio-
remediation techniques which can be carried out by the use of
microorganisms such as bacteria, fungi, or plants. These spe-
cies uptake or absorb the heavy metals or may transform them
by changing their valance state and make them less toxic
(Ayangbenro & Babalola, 2017). The present review article
is focused on the various bioremediation mechanisms which
are used by different microbial systems to overcome the heavy
metal pollution. Herein, the adverse effect of four most com-
Table 1 Contamination sources, uses, and adverse health effects of some heavy metals
Element Containment sources
Natural Anthropogenic
Cd Zn and Pb Mining waste, electroplating, automobile
minerals. exhaust
Phosphate
rocks
Cr Chromite mineral Electroplating, metal alloys, industrial
sewage, anticorrosive products
Cu Sulfides, oxides, Electroplating, metal alloys, domestic and Most uses are based on the
carbonates industrial waste, mining waste,
pesticides
Pb Galena mineral Battery plants, pipelines, coal, gasoline,
pigments
Ni Soils Metal alloys, battery plants, electronics,
industrial waste
Zn Minerals Metal alloys, pigments, electroplating,
(Sulfides, industrial waste, pipelines
oxides,
silicates)
Int Microbiol
mon heavy metal (As, Pb, Hg, and Cr) pollution on human
health and their remediation mechanism is discussed in detail.
Principle of bioremediation
Bioremediation can be defined as the process of elimination of
pollutants from a contaminated site by using microbial sys-
tems. Microorganisms use contaminant as a food and source
of energy (Azubuike et al., 2016). The microorganisms suc-
cessfully break down or transform the complex and toxic con-
taminant into the simpler or less toxic one (Ayangbenro &
Babalola, 2017). The main goal of bioremediation is to stim-
ulate the native microflora in contaminated site by providing
more food and suitable growth conditions so that they can
grow to their full potential and produce more enzymes as
secondary metabolites. These metabolites further efficiently
break down the complex contaminant into simpler ones
(Chen & Wang, 2017). During bioremediation process of the
contaminant, chemical bonds are broken and energy is re-
leased, which is further utilized by the microorganisms for
their growth (Azubuike et al., 2016). The heavy metal
transforming microbial species can be isolated from both aer-
obic and anaerobic environments. The aerobic microorgan-
isms are more frequently utilized for the bioremediation pro-
cesses as compared to the anaerobic microorganisms
(Azubuike et al., 2016). The literature analysis showed that
the total transformation percentage of different heavy metals
by microorganisms is as follows: Cr (27%), Co (20%), Cd
(31%), and Pb (22%) which is contributing about 70% of
Uses Adverse effect on health
Battery plants Respiratory, cardiovascular, renal effects
Pesticides, detergents Mental disturbance, cancer, ulcer,
hypokerotosis
Anemia and other toxicity effect includes
electric conductor indirectly through interaction with other
properties nutrients
Batteries, alloys Neurotoxic
Batteries production, Skin allergies, lung fibrosis, diseases of
catalysts of vegetable oils cardiovascular system
Fertilizers, plastic, pigments Abdominal pain, nausea, vomiting and
diarrhea, irritability, leathery, anemia
Int Microbiol
the total content. Whereas, other metals like Ni (07%), Zn
(05%), and As and Hg are about (18%).
The name of some microorganisms belonging to four major
groups (bacteria, archaea, fungi, and yeast), having metal
transforming capabilities are given in Table 2 (Ayangbenro
& Babalola, 2017; Gupta & Singh, 2017).
Mechanism of microbial bioremediation
Microorganisms transform the heavy metals by using different
strategies. They change the ionic state of metals, which may
affect their solubility, mobility, and bioavailability
(Ayangbenro & Babalola, 2017). The heavy metal bioremedi-
ation can be carried out by mobilization or immobilization
processes, which can be accomplished by following
Table 2 Heavy metal biotransformation potential of some
microorganisms
Organisms Genus/species
Bacteria Arthrobacter sp.
Bacillus cereus
Bacillus cereus strain XMCr-6
Bacillus subtilis
Citrobacter sp.
Cupriavidus metallidurans
Phylum Cyanobacteria sp.
Enterobacter cloacae
Enterobacter cloacae B2-DHA
Kocuria flava
Pseudomonas aeruginosa
Pseudomonas putida
Pseudomonas veronii
Sporosarcina ginsengisoli
Streptomyces sp.
Zoogloea ramigera
Archaea Phylum Crenarchaeota sp.
Phanerochaete chrysosporium
Fungi Aspergillus fumigatus
Aspergillus tereus
Aspergillus versicolor
Gloeophyllum sepiarium
Penicillium chrysogenum
Rhizopus oryzae (MPRO)
Yeast Candida utilis
Hansenula anomala
Rhodotorula mucilaginosa
Rhodotorula rubra GVa5
Saccharomyces cerevisiae
mechanisms, i.e., chelation, oxidation-reduction, pH change,
biosorption, bioaccumulation, immobilization, biomethyla-
tion, or change the organic metallic complex to radionuclides.
The brief description of fundamental metal processing mech-
anisms used by various microorganisms is as follows:
Mobilization
In this process, the redox reaction technique helps to dissolve
the toxic metals and their radionuclides and transforms them
to mineral, organic acids. It also lowers down the pH of the
contaminant. The complete mobilization processes are further
divided into four sub-processes, i.e., enzymatic oxidation, en-
zymatic reduction, complexation, and siderophores. The detail
of these processes is given in subsequent sections.
Enzymatic oxidation
Normally, some inorganic compounds exist in more than one
oxidation states, and it is commonly observed that the higher
oxidation state is less soluble than its lower ionic state; in such
cases, the enzymatic oxidation process (catalyzed by the en-
zymes released from microorganisms) plays a very important
role and enhances the solubility of compounds by oxidizing
the higher state into a lower state. It is one of the important
methods to remove the inorganic species from solution. In this
process, heavy metals lose electrons and get transformed into
the useful or less toxic state. The most common example of
enzymatic oxidation is the oxidation of uranium by
Thiobacillus ferrooxidans, T. thiooxidans from its ore
(Cumberland et al., 2016).
Enzymatic reduction
This process is reverse as compared to the enzymatic oxida-
tion process. The inorganic compounds which have many
oxidation states remained insoluble in their reduced state.
The process of enzymatic reduction proved useful to remove
such elements from solution. In case of in situ bioremediation,
the enzymatic reduction reaction carried out by facultative and
obligate anaerobic microorganisms (Rabus et al., 2016).
Complexation
The complexation is the process of making a complex of the
inorganic metals by the addition of ligand. Due to the metal
complex formation, the toxic inorganic compounds are mobi-
lized and can be easily removed from the solid waste
(Ayangbenro & Babalola, 2017). The major microbial
complexing agents are of two types: (i) low molecular weight
organic acids (citric acid, tricarboxylic acids, and alcohols)
and (ii) high molecular weight ligands, siderophores, and tox-
ic metal-binding compounds. In addition to this, some amino

acids synthesized by certain bacteria also act as complex-
forming agents. It is observed that the complexation of heavy
metal and radionuclides to the microorganisms is highly de-
pendent on pH (Ayangbenro & Babalola, 2017).
Siderophores
During the growth of the microorganism in iron-deficient me-
dia, some microbes produce specific iron chelators which are
also known as siderophores. These siderophores have specific
binding groups such as catecholate, phenolate, or
hydroxamate. Due to the presence of these specific groups,
the complex formation property of these siderophores is in-
creased, and they form various complexes with toxic metals
and hence enhance their solubility (Khan et al., 2017). In
recent years, a large number of siderophores or siderophore-
like compounds have been identified from various biological
systems. It is reported in the literature that siderophores are
specific for Fe (III), but recent studies showed that they can
also complex with other metals and radionuclides (Ahmed &
Holmstrom, 2014).
Immobilization
Ex situ and in situ immobilization techniques are used for
the remediation of the metal-contaminated soil. The ex situ
technique is applied at the highly contaminated area. The
soil of this area is removed from its original place and
stored in a specific place where it can be treated with dif-
ferent microbial systems to immobilize the metal ions pres-
ent in it. Whereas, in case of in situ technique, the metal-
contaminated soil is treated at its original place. In immo-
bilization process, nitrate-nitrogen is converted into organic
nitrogen which is reverse to the mineralization process. The
immobilization process is considered as a biological pro-
cess as it is controlled by several soil bacteria. This process
is affected by soil moisture and soil temperature. The im-
mobilization took place by active (energy-dependent) or
passive (energy-independent) methods. It is observed that
the passive immobilization is nonspecific to metal species
whereas the active immobilization is comparatively slow
and mostly depends on the microbial cell metabolism.
Mostly active immobilization uses specific proteins like
metallothioneins which made a complex with heavy metals.
The heavy metal immobilization is mainly carried out by
precipitation, biosorption, and bioaccumulation. These pro-
cesses are commonly used for the treatment of heavy metal-
contaminated wastewater (Ayangbenro & Babalola, 2017).
The detailed description of various steps of immobilization
process is given in below sections.
Int Microbiol
Precipitation or solidification
The metal ions can be precipitated or solidified in solution or
soil with the help of different methods. The most common
example of precipitation is the sulfate reduction. Sulfate-
reducing bacteria (SRB) are being used in engineering, natural
systems such as constructed wetland to treat metal contami-
nants. SRB remove toxic metals from solution by forming the
metal sulfide precipitates. The solubilities of most of the toxic
metal sulfides are very low, and after forming precipitate, their
toxic effect on the environment almost becomes negligible.
The SRB also create the conditions in solution which favor
the chemical reduction of metals. Microbial transformation of
organo-phosphates to ortho-phosphate can also lead to metal
precipitation through the formation of metal-phosphates, es-
pecially above pH 7. Intracellular phosphates may also immo-
bilize metals (Martinez et al., 2014). Precipitation process can
be carried out both in situ or ex-situ.
Biosorption
Biosorption is the physiochemical process of absorbing the
metal ions by microorganisms such as algae, bacteria, and
fungi. This is an energy-independent process, and these organ-
isms absorb the metal ions and reduced their concentration in
solution (Ayangbenro & Babalola, 2017). Biosorption is much
economical than other metal ion removal strategies. The ca-
pacity of a biomass to recover the metal ions depends on
various physical, chemical, and biological properties. The
common cellulosic natural materials are mostly used as poten-
tial sorbents for heavy metals (Malik et al., 2017). Every
biosorbant organism has specificity for a particular metal ion
(Gupta & Singh, 2017). It is seen that certain extracellular
polymeric substances (EPS), a complex mixture of polysac-
charide, mucopolysaccharides, and proteins are capable to
bind a significant amount of toxic metal ions. In case of
Gram-positive and -negative bacterial species, peptidoglycans
and phosphate groups are main cationic and anionic binding
sites respectively. Whereas, in fungal biomass, the chitin, phe-
nolic polymers and melanin are the most important structural
components which effectively act as heavy metal and radio-
nuclide biosorbent. The biosorbent technique has following
advantages over other known processes, i.e., this is a cheap
process, no drawback, no sludge formation, and as well as this
is a regenerative process. The biosorption can be carried out in
a single- or double-step process. The selection of microbial
species depends upon the concentration of the metal ions in
the solution. In single-step biosorption process, the biosorbent
microbial species are incubated with a medium contaminated
with heavy metal in a mixing tank, and after a specific time
interval (depending on the growth cycle of microbial stains),
the solution is filtered out. Whereas, in double-step
biosorption process, the filtrate of step 1 is again suspended
Int Microbiol
with biosorbent species and filtered after incubation time. The
two-step biosorption method is reported to be much more
effective than single-step method. The biosorption technique
(single/double) is shown in Fig. 1.
Bioaccumulation
Bioaccumulation is an energy-dependent process where the
accumulation of heavy metals is carried out by microorgan-
isms. The bioaccumulation is also a beneficial process like
biosorption. The metals such as mercury, lead, silver, cadmi-
um, nickel, cesium, cobalt, chromium, and uranium are accu-
mulated by the microorganisms (Olaniran et al., 2013). The
microorganism normally accumulates these metals by ion
pumps, ion channels, endocytosis, and lipid permeation
(Satyapal et al., 2016). It is one of the most popular and suc-
cessful technique to restore or accumulate the heavy metals
from the environment. During bioaccumulation, the heavy
metal ions made an inactive complex with other high-
affinity ligands (Satyapal et al., 2016).
Heavy metals as a pollutant and their effect
on health
The heavy metals are those metals whose density is about five
times heavier than water (Sarubbo et al., 2015). These heavy
metals naturally present on the earth in a limited amount, but
due to the uncontrolled human activities (industrial, mining,
smelting, etc.), they get concentrated in a particular site (Das et
al., 2014). This accumulation is harmful to the environment
because these metals generally accumulated in their most sta-
ble oxidation states, i.e., As+3, Pb+2, Hg+2, and Cd+2 which
further react with body bio-molecules to generate extremely
stable bio-toxic compounds which are very difficult to disso-
ciate (Kulshreshtha et al., 2014). The most common heavy
metals which contaminate the soil and water are As, Pb, Hg,
and Cr. The source, toxicity, and harmful effects along with
the brief bioremediation mechanism of these pollutants are
given in subsequent sections.
Fig. 1 Single- and double-step
biosorption process
Arsenic
Arsenic (As) is a trace element which exists in a minute
quantity in our natural diet. Several foods such as seafood,
poultry, grains (especially rice), bread, cereal products,
mushrooms, and dairy products contain As in very low
quantity and concentrations (Cheyns et al., 2017). A min-
ute quantity of As is also used to prepare medicine, but its
exact functions are unknown. The estimated adult daily
intake of arsenic from a typical diet is 12–50 μg, and its
dietary requirement of 12–25 μg per day has been sug-
gested (Mahmood & Malik, 2014). Arsenic is safe when
consumed in a limited amount in the form of various food
components. It is well described that the arsenic present
naturally in foods (organic arsenic) does not cause any
harm at a level of 5 μg/dL (Bräuner et al., 2014). In nature,
it is found in two forms, i.e., organic and inorganic. The
organic form of arsenic is not harmful to human beings, but
its inorganic form is much more harmful, and its poisoning
may cause abdominal pains, destruction of red blood cells
(hemolysis), shock, leuco-melanosis, keratosis, hyperkera-
tosis, dorsum, nonpitting edema, gangrene, and skin cancer
(Parmar & Thakur, 2013). In contaminated groundwater, it
is mostly present in two forms: arsenate (As5+) and arsenite
(As3+) (Paul et al., 2014). The presence of these two forms
(As3+ and As5+) in soil or water mainly depends upon the
physiological conditions (redox condition, pH, etc.) of the
medium. In oxic condition, As5+ is the predominant con-
versely, under anoxic environment. As 3+ is prevalent
which is more toxic and more mobile (Yang et al., 2014).
Microorganism remediates arsenic contamination from the
soil by using mobilization mechanism of bioremediation. The
enzymatic oxidation and reduction are carried out to convert
As3+ to a less toxic arsenic, i.e., As5+. The enzyme arsenic
oxidase catalyzes the arsenic oxidation, whereas As reduction
is carried out by two different ways: cytoplasmic reduction
and periplasmic reduction. The reduced AsIII is either extrud-
ed or accumulated in the intracellular compartments as a free
arsenite or may conjugate with glutathione or other thiols
(Satyapal et al., 2016; Dey et al., 2016).

The common arsenic bio-transforming microorganisms are
as follows: Acinetobacter sp., Brevundimonas sp.,
Pseudomonas sp., Rhizobium sp., Aeromonas sp., and
Penicillium canescens (Layton et al., 2014).
Lead
Lead (Pb) is the second most excessive heavy metal which is
found as the pollutant on the earth. In nature, the presence of
native lead is very rare, and mostly it is associated with the
zinc, silver, and copper ores and extracted along with the
extraction of these metals (Davidson et al., 2014). But with
the development of civilization, the lead utilization increased,
and nowadays it becomes a major pollutant (Khatri & Tyagi,
2015). Lead pollution is higher in the developing world as
compared to the developed world. The lead is being utilized
in gasoline, and a man-made lead-cycle has been developed.
Zhang et al. (Zhang et al., 2015), described that during the
burning of gasoline containing lead in automobile engines
leads to the production of many lead salts such as chlorines,
bromines, and oxides. These lead salts enter the environment
through the automobile exhausts. The larger sized particles
fell on the ground as they exhausted and pollute soil or surface
water, whereas the smaller particles fly away with air and
remained in the atmosphere. Some parts of these small parti-
cles again come in contact with earth from rainwater mixed
with the small particles. The man-made lead-cycle raised the
lead concentration in the environment up to an alarming stage,
and it is much more fatal than natural lead-cycle.
Lead has the most damaging effects on human health, and
it enters into the human body through uptake of food (65%),
water (20%), and air (15%) (Mahmood & Malik, 2014). The
exposure of human beings to the lead is known as lead poi-
soning or lead intoxication. Lead poisoning may be of two
types, i.e., acute (from intense exposure of short duration) or
chronic (from repeated low-level exposure over a prolonged
period). The diagnosis and treatment of lead poisoning depend
on the presence of lead level in the blood which is measured in
micrograms of lead per deciliter of blood (μg/dL) (Parmar &
Thakur, 2013). According to the current guidelines of WHO,
the excessive exposure of lead to a healthy person must be less
than 25 μg/dL for adult and for children, it must be less than
5 μg/dL (WHO, 2010).
Lead bioremediation by various microbial strains is carried
out by using immobilization mechanisms. The microorganism
developed several resistant mechanisms which allow or help
them to withstand the toxic effects of lead. The most common
lead resistance mechanism which is used by the several mi-
crobial systems to remediate the lead from soil or medium is
accumulation, biosorption, and precipitation (Naik & Dubey,
2013). Major Pb biotransforming organisms are Aspergillus
niger, Aspergillus terrus, Aspergillus versicolor, Neurospora
Int Microbiol
crassa, Penicillium canescens, Penicillium chrysogenum,
Penicillium decumbens, Penicillium simplicissimum, and
Saccharomyces cerevisiae (Jebara et al., 2015; Joshi et al.,
2011).
Mercury
Mercury (Hg) is a heavy metal commonly known as quick-
silver having atomic number 80. Hg is the only heavy
metal which exists in its liquid state at standard pressure
and temperature conditions after bromine. Mercury occurs
in deposits throughout the world, mostly as cinnabar (mer-
curic sulfide) (Wang et al., 2012). It is known as one of the
extremely rare elements in Earth’s crust, having an average
crustal abundance by mass of only 0.08 ppm (Ehrlich &
Newman, 2008). Being a rare natural element, it is mobi-
lized by human activities to the environment (aquatic and
terrestrial) since from thousands of years. It is mainly uti-
lized for the extraction of precious metals. Hg is also com-
monly used in certain measurement-related instruments
such as thermometers, barometers, manometers, sphygmo-
manometers, float valves, mercury switches, mercury re-
lays, and fluorescent lamps as well as in paint industries
(Martinez-Finley & Aschner, 2014). There are a variety of
ways by which mercury enters into the environment. The
majority of it emitted in the form of gaseous elemental
mercury, which can be transported far from the emission
source (Driscoll et al., 2013). The other Hg emissions are
in the form of gaseous inorganic ionic mercury forms such
as mercuric chloride. These both forms have a short life-
time, and they start depositing on land or water approxi-
mately 100 to 1000 km away from their emission source.
The ocean tides are another way for long-range mercury
transport (Weber & Thomas, 2017). Hg poisoning has a
number of side effects on human health. The excessive
exposure to Hg causes many kinds of sickness like aller-
gies, the inability to speak, nervous, and renal system-
related disorders (Azimi & Moghaddam, 2013). It can be
bio-remediate from contaminated soil or water by using
both mobilization and immobilization processes. The most
common mechanism used by microorganisms for mercury
biotransformation is changing the valance state of the met-
al ion and make it less toxic by undergoing reduction,
extracellular precipitation, or volatilization (Ramasamy &
Kamaludeen, 2007; Sinha et al., 2009).
The Hg can be transformed by the use following microor-
ganisms: Penicillium canescens, Rhizopus arrhizus,
Shewanella oneidensis, Geobacter sulfurreducens, and
Geobacter metallireducens (Sinha et al., 2009; Dixit et al.,
2015; Wiatrowski et al., 2006).
Int Microbiol

Fig. 2 Genetic engineering in

indigenous bacteria to enhance
bioremediation capability of
microbial strain

Chromium
Chromium (Cr) having atomic number 24, is the 17th most
abundant element in the Earth’s mantle (Bhalerao & Sharma,
2015). Weathering of Cr-containing rocks, industrial effluents,
and soil leaching are the measured reasons of its deposition in
the soil or water bodies (Khatri & Tyagi, 2015). Cr composi-
tion of both water and soil is different. In soil, its concentration
depends on the presence and quantity of Cr-containing rocks
present in that region, whereas in water bodies, Cr concentra-
tion depends on the quantity of industrial effluent mixed with
water (Ertani et al., 2017). In the environment, Cr exists in
various oxidation states like Cr (0), Cr (III), and the hexavalent
Cr (VI) species (Bhalerao & Sharma, 2015). Out of these
states, Cr (0) is the most stable state followed by the Cr (VI)
which exists mostly as chromate (CrO4 2−), dichromate
(Cr2O42 −), and chromium trioxide (CrO3). The CrO3 is the
most toxic state which has high oxidizing potential, along with
high solubility, and it has high mobility across the cell mem-
branes in living organisms. On the other hand, the Cr (III)
exists in the form of oxides, hydroxides, and sulfates, which
are less toxic and relatively insoluble in the water, having low
mobility (Khatri & Tyagi, 2015). In water, the Cr solubility
depends on its pH. Cr (III) is soluble in only in acidic pH
conditions whereas at neutral and alkaline pH, Cr (III) gets
precipitated. The other forms of Cr (VI), i.e., chromate and
dichromate are extremely soluble under all pH (Parmar &
Thakur, 2013; Ertani et al., 2017). Hexavalent chromium (Cr
(VI)) is carcinogenic and mutagenic in nature. The acceptable
Cr concentrations in water are 0.8 and 0.1 μg/dL for Cr (III)
and Cr (VI), respectively. The Cr level in the effluents of Cr
industries varies between 2 and 5 × 10−5 μg/dL which is very
much higher than the recommended concentration (Parmar &
Thakur, 2013). The excessive accumulation of Cr in soil or
water causes various effects on the plant as well as human
health. In case of plants, Cr accumulation causes severe dam-
age to their root, shoot growth, and photosynthesis process
(Rodriguez et al., 2012). Whereas, in case of animals and
microorganisms, it induces hyperexpression of several antiox-
idant enzymes [peroxidase, catalase (CAT), glucose-6-
phosphate dehydrogenase, and superoxide dismutase
(SOD)], which further leads to many kinds of serious

Table 3 Engineered bacteria having modified gene expression and their heavy metal biotransformation capabilities

Heavy metal Bacteria Reference

As E.coli strain (Yuan et al., 2008)

Sphingomonas desiccabilis and Bacillus Idriensis strains (Liu et al., 2011)
Pb Bacillus subtilis BR151; Staphylococcus aureus RN4220 (Bondarenko et al., 2008)
Hg Achromobacter sp. AO22 (Ng et al., 2009)
Acidithiobacillus ferrooxidans strain (Sasaki et al., 2005)
Deinococcus geothemalis (Dixit et al., 2015)
E.coli JM 109 (Zhao et al., 2005)
E.coli MC 1061 (Bondarenko et al., 2008)
Pseudomonas K-62 (Kiyono & Pan-Hou, 2006; Kiyono et al., 2009)
Pseudomonas sp. (Sone et al., 2013)
Cr Methylococcus capulants (Hasin et al., 2010)
P. putida (Ackerley et al., 2004)
Cd B. subtilis BR151 (pT0024) (Ivask et al., 2011)
E.coli strain (Freeman et al., 2005)
Ni E.coli SE5000 (Deng et al., 2005)
E.coli strain (Freeman et al., 2005)
P. fluorescens 4F39 (Lopez et al., 2002)

Table 4 Major advantages and limitations of microbial remediation
Advantages
Is a natural process and is therefore perceived by the public.
Is useful for the complete destruction of a wide variety of contaminants.
The microbes did not produce any secondary pollutant during
biodegradation.
Instead of transferring contaminants from one environmental medium to Are often highly specific and these processes depend upon microbial
another, for example, from land to water or air, the complete destruction
of target pollutants is possible.
Can often be carried out on site, often without causing a major disruption It is difficult to extrapolate (deduce) from bench and pilot-scale studies to
of normal activities.
Can prove less expensive than other technologies that are used for
cleanup of hazardous waste.
disorders. Chromium can be bio-remediate by several micro-
bial species by using different mechanisms. The microbial
species carry out the metabolic reduction of Cr (VI) to Cr
(III) or either via extracellular or intracellular precipitation
(Jouety et al., 2015).
The major chromium transformers are Pantoea sp.,
Bacillus circulans, Bacillus megaterium, Bacillus coaglans,
Zoogloea ramigera, Streptomyces nouresei, Aeromonas
caviae, Pseudomonas sp., and Staphylococcus xylosus
(Malaviya & Singh, 2014; Aryal & Liakopoulou-Kyriakides,
2015).
Genetic engineering for improved microbial
remediation of heavy metal pollution
The microbial strains whose genetic makeup is reformed with
the help of molecular biology tools to enhance the biodegra-
dation or biotransformation capabilities are known as geneti-
cally engineered microorganisms (GEM’s) (Zhang et al.,
2015). The GEM’s development is mainly achieved by four
principal approaches, i.e., (i) modification of enzyme specific-
ity and affinity, (ii) construction and regulation of specific
pathways, (iii) development of bioprocess for remediation
and its monitoring and control (iv) Use and applications of
biosensors in chemical sensing, toxicity reduction, and end-
point analysis (Gupta & Singh, 2017). On the bases of these
approaches, genetically engineered microorganisms are gen-
erated to minimize the heavy metal pollution. The GEMs can
be produced by making changes in different genes. Singh et al.
(Singh et al., 2011), described the genetically engineered bac-
terial gene for heavy metal degradation or biotransformation
in details (Fig. 2).
The engineered microorganisms have enhanced capabili-
ties to transform the heavy metals to their corresponding less
harmful states. A number of microbial systems are engineered
till date; a brief detail of these microbial systems is given in
Int Microbiol
Limitations
Is limited to those compounds that are biodegradable. Not all compounds
are susceptible to rapid and complete degradation.
The products of biodegradation may be more persistent or toxic than the
parent compound.
populations, suitable environmental growth conditions, and appropriate
levels of nutrients and contaminants.
full scale field operations.
Bioremediation often takes longer time than other treatment options.
Table 3 (Singh et al., 2011). Application of genetically
engineered microbial system for the remediation of heavy
metal pollution is a cheap and safe alternative due to the se-
lective nature of GE microbes which pose a very few health
hazards as compared to the other physicochemical methods.
Advantages and limitations of microbial
remediations
Microorganisms play a very important role in our daily life.
They remediate or biotransform the contaminant or waste by
using it as the main source of carbon and energy. The major
advantages and limitations of microbial remediations are tab-
ulated in Table 4.
Conclusion
Heavy metal pollution is widespread pollution all over the
world. It is observed that many natural and industrial activities
generate metal ions which reach to our environment (soil and
water) and disturb their natural composition. In last one de-
cade, a large number of studies have been conducted to lower
down the heavy metal pollution. Currently, many research
groups are working on the discovery of new pathways and
techniques to lower down this pollution. Further, research
groups’ investigations are aimed at the identification of new
potential microbial species, which can transform heavy metal
faster as well as up to the large content. Also, there is need to
find out the new mechanisms of heavy metal bioremediation
and use of genetic engineering techniques to enhance the bio-
remediation potential. The studies described that bioremedia-
tion cost is still high, so there is a need to generate such type of
technology or methodology which further reduces the biore-
mediation cost. Researchers are also working with transgenic
or genetically modified microorganisms to improve the
Int Microbiol
biodegradation rate. The genetically modified microorgan-
isms can be successfully employed to remediate the heavy
metal pollution from the environment, but unfortunately, these
transgenic showed their maximum potential under controlled
(lab) conditions. To enhance the remediation rate of heavy
metal from containment area, we have to work towards the
stability of transgenic under in situ conditions.
Acknowledgments All content of the review article is fully
acknowledged.
Funding information This study was funded by the National Science
Foundation of China (31670117), the Science & Technology Project of
Guangdong Province, China (2014A020217017), and the Guangdong
Natural Science Foundation, China (2014A030313475).
References
Ackerley DF, Gonzalez CF, Keyhan M, Blake R 2nd, Matin A (2004)
Mechanism of chromate reduction by the Escherichia coli protein,
NfsA, and the role of different chromate reductases in minimizing
oxidative stress during chromate reduction. Environ Microbiol 6:
851–860
Ahmed E, Holmstrom SJM (2014) Siderophores in environmental re-
search: roles and applications. Microbial Biotechnol 7:196–208
Aryal M, Liakopoulou-Kyriakides M (2015) Bioremoval of heavy metals
by bacterial biomass. Environ Monit Assess 187(4173):1–26
Ayangbenro AS, Babalola OO (2017) A new strategy for heavy metal
polluted environments: a review of microbial biosorbents. Int J
Environ Res Public Health 14(1):94
Azimi S, Moghaddam MS (2013) Effect of mercury pollution on the
urban environment and human health. Environ Ecology Res 1(1):
12–20
Azimi A, Azri A, Rezkazemi M, Ansarpour M (2017) Removal of heavy
metals from industrial wastewaters: a review. Chem Bio Eng
Reviews 4:37–59
Azubuike CC, Chikere CB, Okpokwasili GC (2016) Bioremediation
techniques—classification based on site of application: principles,
advantages, limitations and prospects. World J Microbiol Biotechnol
32(11):180
Bhalerao SA, Sharma AS (2015) Chromium: as an environmental pollut-
ant. Int J Curr Microbiol App Sci 4(4):732–746
Bondarenko O, Rolova T, Kahru A, Ivask A et al (2008) Bioavailability
of Cd, Zn and Hg in soil to nine recombinant luminescent metal
sensor bacteria. Sensors 8:6899–6923
Bräuner EV, Nordsborg RB, Andersen ZJ, Tjønneland A, Loft S,
Raaschou-Nielsen O (2014) Long-term exposure to low-level arse-
nic in drinking water and diabetes incidence: a prospective study of
the diet, cancer and health cohort. Environ Health Perspect 122:
1059–1065
Chen H, Wang L (2017) Chapter 5—microbial cell refining for biomass
conversion. Technologies for Biochemical Conversion of Biomass
101–135
Cheyns K, Waegeneers N, de Wiele TV, Ruttens A (2017) Arsenic release
from food stuffs upon food preparation. J Agri Food Chem 65(11):
2443–2453
Chibuike GU, Obiora SC (2014) Heavy metal polluted soils: effect on
plants and bioremediation methods. Applied and Environmental
Soil Science Article ID 752708: 1–12
Cumberland SA, Douglas G, Grice K, Moreau JW (2016) Uranium mo-
bility in organic matter-rich sediments: a review of geological and
geochemical processes. Earth Sci Rev 159:160–185
Das S, Raj R, Mangwani N, Dash HR, Chakraborty J (2014) 2-heavy
metals and hydrocarbons: adverse effects and mechanism of toxic-
ity. Microbial Biodegradation and Bioremediation 23–54
Davidson A, Ryman J, Sutherland CA, Milner EF, Kerby RC, Teindl H,
Melin A, Bolt HM (2014) Lead. Ullmann’s Encyclopedia of
Industrial Chemistry doi:https://doi.org/10.1002/14356007.a15_
193,
Deng X, Li QB, Lu YH, He N, Jiang J (2005) Genetic engineering of
Escherichia coli SE5000 and its potential for Ni2+ bioremediation.
Process Biochem 40:425–430
Dey U, Chatterjee S, Mondal NK (2016) Isolation and characterization of
arsenic-resistant bacteria and possible application in bioremediation.
Biotechnol Rep 10:1–7
Dixit R, Wasiullah, Malaviya D, Pandiyan K, Singh UB, Sahu A, Shukla
R, Singh BP, Rai JP, Sharma PK, Lade H, Paul D (2015)
Bioremediation of heavy metals from soil and aquatic environment:
an overview of principles and criteria of fundamental processes.
Sustainability 7(2):2189–2212
Driscoll CT, Mason RP, Chan HM, Jacob DJ, Pirrone N (2013) Mercury
as a global pollutant: sources, pathways, and effects. Environ Sci
Technol 47:4967–4983
Ehrlich HL, Newman DK (2008) Geomicrobiology. CRC Press. 265,
Geomicrobiology of Mercury
Ertani A, Mietto A, Borin M, Nardi S (2017) Chromium in agricultural
soils and crops: a review. Water Air Soil Pollut 228:190
Freeman JL, Persans MW, Nieman K, Salt DE (2005) Nickel and cobalt
resistance engineered in Escherichia coli by over expression of ser-
ine acetyl transferase from the nickel hyper accumulator plant
Thlaspi goesingense. Appl Environ Microbiol 71:8627–8633
Govind M (2014) Heavy metals causing toxicity in animals and fishes.
Res J Animal Veterinary and Fishery Sci 2(2):17–23
Gupta S, Singh D (2017) Role of genetically modified microorganisms in
heavy metal bioremediation. Springer Nature Singapore Pvt. Ltd. R.
Kumar et al. (eds.), Advances in environmental biotechnology DOI
https://doi.org/10.1007/978-981-10-4041-2_12
Hansda A, Kumar V, Anshumali V, Usmani Z (2014) Phytoremediation
of heavy metals contaminated soil using plant growth promoting
rhizobacteria (PGPR): a current perspective. Recent Res in Sci &
Tech 6(1):131–134
Hasin AA, Gurman SJ, Murphy LM, Perry A, Smith TJ, Gardiner PE
(2010) Remediation of chromium (VI) by a methane-oxidizing bac-
terium. Environ Sci Technol 44:400–405
Ivask A, Dubourguier HC, Pollumaa L, Kahru A (2011) Bioavailability
of Cd in 110 polluted top soils to recombinant bioluminescent sensor
bacteria: effect of soil particulate matter. J Soils Sediments 11:231–
237
Jebara SH, Abdelkerim S, Fatnassi IC, Chiboub M, Saadani O, Jebara M
(2015) Identification of effective Pb resistant bacteria isolated from
Lens culinaris growing in lead-contaminated soils. J Basic
Microbiol 55:346–353
Joshi PK, Swarup A, Maheshwari S, Kumar R, Singh N (2011)
Bioremediation of heavy metals in liquid media through fungi iso-
lated from contaminated sources. Indian J Microbiol 51:482–487
Jouety NT, Sayel H, Bahafid W, El Ghachtouii N (2015) Mechanisms of
hexavalent chromium resistance and removal by microorganisms.
Rev Environ Contam Toxicol 233:45–69
Khan A, Singh P, Srivastava A (2017) Synthesis, nature and utility of
universal iron chelator–Siderophore: a review. Microbiol Res, in
press 212-213:103–111. https://doi.org/10.1016/j.micres.2017.10.
012
Khatri N, Tyagi S (2015) Influences of natural and anthropogenic factors
on the surface and groundwater quality in rural and urban areas.
Front Life Sci 8(1):23–39

Kiyono M, Pan-Hou H (2006) Genetic engineering of bacteria for envi-
ronmental remediation of mercury. J Health Sci 52:199–204
Kiyono M, Sone Y, Nakamura R, Pan-Hou H, Sakabe K (2009) The Mer
E protein encoded by transposon Tn21 is a broad mercury transport-
er in Escherichia coli. FEBS Lett 583:1127–1131
Kulshreshtha A, Agrawal R, Barar M, Saxena S (2014) A review on
bioremediation of heavy metals in contaminated water. IOSR J
Environ Sci Toxicol Food Tech (IOSR-JESTFT) 8(7) I):44–50
Layton AC, Chauhan A, Williams DE, Mailloux B, Knappett PS,
Ferguson AS, McKay LD, lam MJ, Matin Ahmed K, Van Green
A, Sayler GS (2014) Metagenomes of microbial communities in
arsenic- and pathogen-contaminated well and surface water from
Bangladesh. Genome Announc 2(6):e01170–e01114
Liu S, Zhang F, Chen J, Sun G (2011) Arsenic removal from contaminat-
ed soil via biovolatilization by genetically engineered bacteria under
laboratory conditions. J Environ Sci 23(9):1544–1550
Lopez A, Lazaro N, Morales S, Marques AM (2002) Nickel biosorption
by free and immobilized cells of Pseudomonas fluorescens4F39: a
comparative study. Water Air Soil Pollut 135:157–172
Mahmood A, Malik RN (2014) Human health risk assessment of heavy
metals via consumption of contaminated vegetables collected from
different irrigation sources in Lahore, Pakistan. Arab J Chem 7(1):
91–99
Malaviya P, Singh A (2014) Bioremediation of chromium solutions and
chromium-containing wastewaters. Crit Rev Microbiol 42(4):607–
633
Malik DS, Jain CK, Yadav AK (2017) Removal of heavy metals from
emerging cellulosic low-cost adsorbents: a review. Appl Water Sci
7:2113–2136
Martinez RJ, Beazley MJ, Sobecky PA (2014) Phosphate-mediated reme-
diation of metals and radionuclides. Advances in Ecology Article ID
786929, 1–14
Martinez-Finley EJ, Aschner M (2014) Recent advances in mercury re-
search. Current Environmental Health Reports 1(2):163–171
Naik MM, Dubey SK (2013) Lead resistant bacteria: lead resistance
mechanisms, their applications in lead bioremediation and biomon-
itoring. Ecotoxicol Environ Saf 98:1–7
Ng SP, Davis B, Polombo EA, Bhave M (2009) Tn5051 like mer con-
taining transposon identified in a heavy metal tolerant strain
Achromobacter sp. AO22 BMC Res Notes 7:2–38
Olaniran AO, Balgobind A, Pillay B (2013) Bioavailability of heavy
metals in soil: impact on microbial biodegradation of organic com-
pounds and possible improvement strategies. Int J Mol Sci 14(5):
10197–10228
Parmar M, Thakur LS (2013) Heavy metal Cu, Ni and Zn: toxicity, health
hazards and their removal techniques. Int J Plant Animal and
Environ Sci 3(3):143–157
Paul D, Poddar S, Sar P (2014) Characterization of arsenite-oxidizing
bacteria isolated from arsenic-contaminated groundwater of West
Bengal. Journal Of Environmental Science And Health Part A
49(13):1481–1492
Rabus R, Boll M, Heider J, Meckenstock RU, Buckel W, Einsle O,
Ermler U, Golding BT, Gunsalus RP, Kroneck P, M H, Krüger M,
Lueders T, Martins BM, Musat F, Richnow HH, Schink B, Seifert J,
Szaleniec M, Treude T, Ullmann GM, Vogt C, von Bergen M,
Wilkes H (2016) Anaerobic microbial degradation of hydrocarbons:
from enzymatic reactions to the environment. J Mol Microbiol
Biotechnol 26(1–3):5–28
Ramamurthy AS, Memarian R (2012) Phytoremediation of mixed soil
contaminants. Water Air & Soil Pollut 223(2):511–518
Ramasamy K, Kamaludeen, Banu SP (2007) Bioremediation of metals:
microbial processes and techniques. In: Singh N, Tripathi R, editors.
Int Microbiol
Environmental bioremediation technologies. Springer Berlin
Heidelberg; 2007. p. 173–187
Rodriguez E, Santos C, Azevedo R, Mountinho-Pereira J, Correia C, Dias
MC (2012) Chromium (VI) induces toxicity at different photosyn-
thetic levels in pea. Plant Physiol & Biochem 53:94–100
Sarubbo LA, Rocha RB Jr, Luna JM, Rufino RD, Santos VA, Banat IM
(2015) Some aspects of heavy metals contamination remediation
and role of biosurfactants. Chem Ecol 31(8):707–723
Sasaki Y, Minakawa T, Miyazaki A, Silver S, Kusano T (2005)
Functional dissection of a mercuric ion transporter MerC from
Acidithiobacillus ferrooxidans. Biosci Biochem Biotech 69:1394–
1402
Satyapal GK, Rani S, Kumar M, Kumar N (2016) Potential role of arsenic
resistant bacteria in bioremediation: current status and future pros-
pects. J Microb Biochem Technol 8:3
Sharma B, Singh S, Siddiqi NJ (2014) Biomedical implications of heavy
metals induced imbalances in redox systems. BioMed Res Int
Article ID 640754:1–26
Singh JS, Abhilash PC, Singh HB, Singh RP, Singh DP (2011)
Genetically engineered bacteria: an emerging tool for environmental
remediation and future research perspectives. Gene 480:1–9
Sinha RK, Valani D, Sinha S, Singh S, Herat S (2009) Bioremediation of
contaminated sites: a low-cost nature’s biotechnology for environ-
mental clean up by versatile microbes, plants & earthworms. Solid
Waste Management and Environmental Remediation 1–72
Sone Y, Mochizuki Y, Koizawa K, Nakamura R, Pan-Hou H, Itoh T,
Kiyono M (2013) Mercurial-resistance determinants in
Pseudomonas strain k-62 plasmid pmr68. AMB Express 3:41
Tang W, Shan B, Zhang H, Zhang W, Zhao Y, Ding Y, Rong N, Zhu X
(2014) Heavy metal contamination in the surface sediments of rep-
resentative limnetic ecosystems in eastern China. Sci Rep 4:7152
Uqab B, Mudasir S, Nazir R (2016) Review on bioremediation of pesti-
cides. J Bioremediat Biodegrad 7(3):1–5
Wang J, Feng X, Anderson CWN, Xing Y, Shang L (2012) Remediation
of mercury-contaminated sites—a review. J Hazardous Mat 221-
222:1–18
Weber T, Thomas M (2017) Influence of ocean tides on the general ocean
circulation in the early Eocene. Paleoceanography 32:553–570
WHO (2010) Preventing disease through healthy environments.
Exposure to lead: a major public health concern
Wiatrowski HA, Ward PM, Barkay T (2006) Novel reduction of
mercury(II) by mercury-sensitive dissimilatory metal reducing bac-
teria. Environ Sci Technol 40(21):6690–6696
Xu R, Chen M, Fang T, Chen J (2017) A new method for extraction and
heavy metals removal of abalone visceral polysaccharide. J Food
Process Preserv 41:e13023
Yang L, Li X, Chu Z, Ren Y, Zhang J (2014) Distribution and genetic
diversity of the microorganisms in the biofilter for the simultaneous
removal of arsenic, iron and manganese from simulated groundwa-
ter. Bioresour Technol 156:384–388
Yuan CG, Lu XF, Qin J, Rosen BP, Le XC (2008) Volatile arsenic species
released from Escherichia coli expressing the AsIII S-adenosyl me-
thionine methyl transferase gene. Environ Sci Technol 42:3201–
3206
Zhang R, Wilson VL, Hou A, Meng G (2015) Source of lead pollution, its
influence on public health and the countermeasures. Int J of Health,
Animal science and Food safety 2:18–31
Zhao XW, Zhou MH, Li QBLYH, He N, Sun DH, Deng X (2005)
Simultaneous mercury bioaccumulation and cell propagation by ge-
netically engineered Escherichia coli. Process Biochem 40:1611–
1616