DAYANANDA SAGAR COLLEGE OF

BIOLOGICAL SCIENCES
(Shivage Malleshwara Hills, Kumaraswamy Layout,
Bangalore-560 078)

Heavy-metals & their effects

on Microorganisms
Submitted to: Mrs. Mamtha mam
Department of Microbiology

Submitted by: Bhuvanesh SK

4th Semester M.Sc. Microbiology
 What is heavy metals ?
• Heavy metals are  natural components of the Earth's crust, with a
specific gravity that is at least 5 times the specific gravity of water &
that has a relatively high density and is toxic or poisonous at low
concentrations. E.g. mercury (Hg), cadmium (Cd), arsenic (As),
chromium (Cr), thallium (Tl), and lead (Pb).
• They cannot be degraded or destroyed
• As trace elements, some heavy metals (e.g. copper, selenium, zinc)
are essential to maintain the metabolism of the cell.
• At higher concentrations they can lead to poisoning.
• Essential Metals play an integral role in the life processes of
microorganisms; serve as micronutrients as components of various
enzymes and for regulation of osmotic pressure.
• Nonessential metals have no biological role and are potentially toxic
to microorganism; through the displacements of essential metals from
their native binding sites or through legend interactions.
• At high levels, both essential and nonessential metals can damage
cell membranes, alter specificity, disrupt cellular functions and
damage the structure of DNA.
• Metals in the environment can be divided
into two classes:
(1) Bioaviable (soluble, nonsorbed and
mobile) and
(2) Nonbioavailable (precipitated,
complexed, and thus toxic to biological
systems).
 Different sources of heavy metals
Heavy metals Sources
As Semiconductors, petroleum refining, wood preservatives, animal feed
additives, coal power plants, herbicides, volcanoes, mining and
smelting
Cu Electroplating industry, smelting and refining, mining, biosolids
Cd Geogenic sources, anthropogenic activities, metal smelting and refining,
fossil fuel burning, application of phosphate fertilizers, sewage sludge
Cr Electroplating industry, sludge, solid waste, tanneries
Pb Mining and smelting of metalliferous ores, burning of leaded gasoline,
municipal sewage, industrial wastes enriched in Pb, paints
Hg Volcano eruptions, forest fire, emissions from industries producing caustic
soda, coal, peat and wood burning
Se Coal mining, oil refining, combustion of fossil fuels, glass manufacturing
industry, chemical synthesis (e.g., varnish, pigment formulation)
Ni Volcanic eruptions, land fill, forest fire, bubble bursting and gas exchange
in ocean, weathering of soils and geological materials
Zn Electroplating industry, smelting and refining, mining, biosolids
Position of heavy metals in Mendeleef table

1. Macrobiogenetic elements 3. Toxic elements

2. Microbiogenetic elements 4. Elements with Chemoterapeutical effect

 Heavy Metal Toxicity
• Effects of heavy metals can be drawn on
i) disruptions on a vast assortment of metabolic processes;
ii) alterations on a balance of pro-oxidant and antioxidant systems;&
iii) competitions with nutrient trace elements for binding sites on essential
metalloenzymes, receptors, metal-binding transporter and storage
systems.
e.g.
• Act as pro-oxidant which catalyzes the production of peroxides and
enhances the subsequent formation of hydroxy radicals and lipid
peroxides, and form highly reactive oxygen species.

• Act as anti-oxidatant result in inhibition of superoxide dismutase

(SOD) and catalase action

• Interact with free sulfhydryl groups,and inhibit enzymes involved in

various metabolism;

• Formation of free radicals, which interfere in protein, nucleic acid, and

energy metabolism
 Classification of naturally occurring metals
by toxicity

• ORDER OF SENSITIVITY to metal ions of heterotrophic microorganism is

Ag+ ≥ Cu2+ = Ni2 > Ba2+ = Cr6+ = Hg2+ > Zn2+ = Na+ = Cd2+ = Pb2+
• The 3 most pollutants heavy metals are Lead, Cadmium, and Mercury.
 Heavy Metal Toxicity Cont..
• Structural similarity of divalent metal cations (e.g. Mn2+, Fe2+, Co2+, Ni2+,
Cu2+ and Zn2+) ;chromate resembles that of sulfate, and arsenate to PO4.

¤ Two types of uptake systems for metal ions.

1) fast, unspecific, and driven by the chemiosmotic gradient across the cytoplasmic
membrane of bacteria. Since this mechanism is used by a variety of
substrates, it is constitutively expressed.
2) high substrate specificity, is slower, often uses ATP hydrolysis as the energy
source and is only produced by the cell in times of need, starvation or a special
metabolic situation; they are inducible.

¤ The governing sorption mechanism's for heavy metals in microorganisms depend

largely on:
1. Cell viability (passive or active sorption),
2. Microbial species (eucaryote: gram-positive/negative, archaebacteria),
3. Heavy metal levels in solution (essential trace element or toxic compound),
4. Chemical composition and pH of the liquid phase,
5. Temperature, and
6. Growth phase of the microorganism.
• Ether and ester linkage
Means by which bacteria react in presence of Metals(M2+) in medium
Metal-binding
M 2+
proteins
n
r p tio
so M M
bio _
_
t ive _ M
N a M2+_
_
_
_
M2+

M(ox.)
M-CH2 e¯
Metal
Enzymatic M(red)
precipitation
Transformation

H2S
M(red) M(ox.) HPO42¯

+M2+
 Heavy Metal Resistance
• Because metal ions cannot be degraded or
modified like toxic organic compounds, there are
six possible mechanisms for a metal resistance
system:
(1) exclusion by permeability barrier;
(2/3) intra- and extra-cellular sequestration;
(4) active efflux pumps;
(5) enzymatic reduction; and
(6) reduction in the sensitivity of cellular targets to
metal ions
 Arsenic
• Appears in Group V of the periodic table
• Semi metallic
• Exists in both organic and inorganic form
• Four oxidation states: -3, 0, +3, +5
• The elemental form of arsenic is As (0)
• Major inorganic forms are As+5 & As+3
• Organic forms : Monomethylarsonic acid (MMA),
Dimethylarsinic acid (DMA), and
Trimethylarsine oxide (TMAO)
• In an aerobic environment, As (V) is dominant and is co-precipitate with or
absorb into iron oxyhydroxides under acidic and moderately reducing
conditions.
• Protonated forms are, H3As03, H2As03-, and HAs032-.
• Under extreme reducing conditions, elemental arsenic and arsine AsH 3 may
be present.
• Does not from complexes with anions such as Cl- and S042-.
• Methylated form of arsine are, dimethyl arsine HAs (CH3)2 and
trimethylarsinic acid (CH3) As02H2 and dimethylarsenic acid (CH3)2 As02H.
Strucutre of arcenic compounds
 Toxicity

• It replace phosphate in many biochemical

reactions
• Presence of arsenic leads to arsenolysis
and depletion of ATP.
• Its Organic forms interact with specific
functional groups in enzymes, receptors,
or coenzymes.
Transport of and resistance to As(V) in Escherichia coli
As(V) As(III)

PhoE PhoE OM

As(V) PR
As(III)

PstABC Pit ArsB CPM

ArsA
As(V)
As(III)

Regulation :- CT

arsR D A B C ArsC
Arsenic metabolism
As(V) enter through phosphate
transport systems such as the
Pst ABC ATPase.

As(V) is reduced to As(III) by

ArsC, a Grx–GSH-linked
enzyme.

The resulting As(III) is pumped

out of the cells by the
ArsA/ArsB ATPase.
The arsRDABC operon is
regulated by the ArsR
repressor protein and ArsD
co-regulator protein.
Transport and metabolism of arsenic in Bacillus subtilis

As (V) enters the cell via the phosphate uptake system (Pit).
As (III) is thought to be taken up by glycerol transporters (GlpF).

• Three products of the

ars-operon have
known functions.
(1)ArsC reduces
As (V) to As (III),
(2)ArsB actively
pumps As (III) out of
the cell,while
(3)ArsR regulates the
expression of the
operon.
• The ars-operon contains four
genes: arsR, yqcK, arsB, and arsC.
 The Reaction Mechanism Of ArsC

1) an oxyanion [As(V), phosphate, or sulfate] is bound noncovalently to the enzyme; a

covalent thiarsahydroxy intermediate forms with the thiolate of Cys12.
2) a glutathionylated intermediate is formed
3) is then reduced to novel ArsC-S-As+-O intermediate
4) In the final step,this enzyme-As(III) complex dissociates to release free As(III).
 3 families
of
Arsenate
Reductases

• Reductases involved in As(V) detoxification have apparently evolved independently at least

three times. One family is typified by the E. coli plasmid R773 ArsC that uses Grx and GSH
as reductants.
• A second family represented by the S. aureus plasmid pI258 ArsC uses thioredoxin (Trx) as
a reductant and is related to a family of lmwPTPases.
• Acr2p is the only eukaryotic arsenate reductase so far identified. It belongs to the
superfamily of PTPases that includes the Cdc25a cell-cycle phosphatase.
 CONCLUSION

• All these mechanisms aim at reducing the

intracellular concentration of a metal so as to
protect the cellular targets
• Free metal ions as they are likely to be more
toxic than the bound ones
• Adaptation to live in such conditions
• Mechanisms of toxic ion resistance may help in
basic understanding of the physiology of the cell
and in enhancing the ability of microorganisms
to extract deleterious ions from the environment
DISCUSSION
 REFERENCE
• Agency for Toxic Substances and Disease Registry, ATSDR. 1999.
http://www.atsdr.cdc.gov.
• Adebowale Adeniji (2004). Bioremediation of Arsenic, Chromium, Lead, and
Mercury
• Lesle Lai. (2003): The carcinogenicy, Mutagenicity, and Toxicity of Arsenic and
Cadmium, SS Biology, MIT
• Bushra Muneer, (2005). Role of Microorganism in Remediation of Heavy Metal in
the Wastewater of Tanneries
• Mohammad Iqbal Lone, Zhen-li He, Peter J. Stoffela, Xiao-e Yang, (2008).
Phytoremediation of heavy metal polluted soils and water:Progresses and
perspectives Lone et al. / J Zhejiang Univ Sci B 2008 9(3):210-220
• Riina Turpeinen(2002). Interactions between metals, microbes and plants –
Bioremediation of arsenic and lead contaminated soils
• Turpeinen, R., Pantsar-Kallio, M., Häggblom, M., Kairesalo, T. Influence of
microbes on mobilization, toxicity and biomethylation of arsenic in soil. The
Science of the Total Environment 236 (1999) 173-180.
• CHAALAL Omar; ZEKRI Adulrazag Y.; ISLAM Rafiq. Uptake of heavy metals by
microorganisms: An experimental approach 2005, vol. 27, no1-2, pp. 87-100 [14
page(s) (article)] (25 ref.)
• http://www.cob.lu.se/arsenic/proj_descr.html
• http://www.bionet.schule.de/schulen/novaky/heavy_metals/en/hm01.htm
 Thank You

For any query please write to bhuvanesh611@gmail.com