Heavy metals:

Fe, Mo, Mn Low toxicity

Zn, Ni, Cu, V, Co, Average toxicity

W, Cr
As, Ag, Sb, Cd, Hg, High toxicity
Pb, U
 Heavy metals:
• Sources: mining operations, refining ores, sludge
disposal, fly ash from incinerators, the processing of
radioactive materials, metal plating, or the
manufacture of electrical equipment, paints, alloys,
batteries, pesticides or preservatives.
• Major lead pollution is through automobiles and
battery manufacturers.
• For zinc and chromium the major application is in
fertilizer and leather tanning respectively.
 Removal of metal ions from aqueous
stream:
• Reverse Osmosis
• Electrodialysis:
• Ultrafiltration:
• Ion-Exchange:
• Chemical precipitation:
• Phytoremediation
• BIOSORPTION
– Biosorption can be defined as the ability of biological materials to
accumulate heavy metals through metabolically mediated or physico-
chemical pathways of uptake 
– Biosorption is a physiochemical process that occurs naturally in
certain biomass which allows it to passively concentrate and bind
contaminants onto its cellular structure.
 Advantage of BIOSORPTION over other
methods:
•   Low cost;
•   High efficiency;
•   Minimisation of chemical and or biological
sludge;
• No additional nutrient requirement;
•   Regeneration of biosorbent; and
•   Possibility of metal recovery.
 Biosorption vs. Bioaccumulation
Microorganisms uptake metal either actively (bioaccumulation)
and/or passively (biosorption).
BIOSORPTION BIOACCUMULATION
• It is a metabolically passive •  It is an active metabolic
process i.e. it does not process driven by energy
require energy. from respiration.
• It is more active in dead cells. • It is active in living cells.
• It is faster and produces
higher concentration of
metals.
• It is a slower process.
• Metal affinity is high under • Toxicity affects metal
favorable conditions. uptake by living cells, but
• Temperature tolerance is may have a high uptake.
modest • Usually affected by low
• It is a reversible process temperature.
• • It is only partially reversible.
• The biosorption process involves a solid phase
(sorbent or biosorbent; biological material) and a
liquid phase (solvent; normally water) containing a
dissolved species to be sorbed (sorbate, metal ions).
• Due to higher affinity of the sorbent for the sorbate
species, the latter is attracted and bound there by
different mechanisms.
• The process continues till equilibrium is established
between the amount of solid-bound sorbate species
and its portion remaining in the solution.
• The degree of sorbent affinity for the sorbate
determines its distribution between the solid and
liquid phases.
 Biosorbent material
• Strong biosorbent behaviour of certain micro-
organisms towards metallic ions is a function of the
chemical make-up of the microbial cells. This type of
biosorbent consists of dead and metabolically
inactive cells.

• Some microbes may collect a broad range of metals

while some are specific to certain metals.
 Biosorbents
• Bacteria: the walls of bacteria are efficient
metal chelators.
• Metal binding may be at least a two-stage process ---
first involving interaction between metal ions and
reactive groups followed by inorganic deposition of
increased amounts of metal.
• The carboxyl groups of glutamic acid of
peptidoglycan is the major group for metal
deposition.
• Eg: Staphylococcus saprophyticus are used for
removal of Cr, Pb and Cd from industrial wastewater.
Algae : use of algae is becoming more
advantageous than others
• Algae have low nutritional requirements as they are
autotrophic and produce huge biomass
• And ,compared to bacteria and fungi produce less toxic
materials.
• Binding depends on : ionic charge of metal ions, chemical
composition of metal soln. and algal sp.
• Metabolism-independent accumulation of metals.
• many potential binding sites occur in algae cell walls, which
include polysaccharides, cellulose, uronic acid and proteins.

• Eg: uptake of Pb by Chlorella vulgaris, Cr (VI) by green algae

spirogyra , Cu by broen seaweed Sargassam sp.
Fungi and Yeast:
• Fungal materials have high percentage of cell wall
material that shows high metal binding capacity.
• Metabolism-independent binding of metal ions is rapid
and in large amounts.
• A variety of ligands may be involved including,
phosphate, hydroxyl and sulphydryl groups. carboxyl,
amino

• Some sp like Aureobasidium pullanans and

Cladosporium resinae show two phase metal uptake.
• Aspergillus niger is found to remove Pb, Cu and Cd.
• Sacchromyces cerevisiae for removal of Pb, Fe,Cu, Cr.
etc.
Agricultural wastes:
• variety of by-products are incorporated, e.g., wool,
olive cake, sawdust, pine needles, almond, and coal.
• Either protein-based animal fibres, or cellulose-based
plants fibres bear many amino, carboxylic or
hydroxyl functional groups which are used for metal
uptake.
• These are inexhaustible , cheap and non-hazardous
which can be easily disposed by incineration.
 Mechanism :
• The mechanism for biosorption may be complex :

– Ion exchange
– Chelation
– Adsorption by physical forces
– Entrapment in inter and intrafibrilliar capillaries and
spaces of the structural polysaccharide network as a
result of the concentration gradient.
– Diffusion through cell walls and membranes .
 Mechanism :
I ) According to the dependence on the cell's
metabolism, biosorption mechanisms can be divided
into……

Metabolism dependent: When transport of metals

occur across the cell membrane, it leads to
intracellular accumulation, this is dependent on cell’s
metabolism.
 Takes place only with viable cells.
 It is often associated with an active defence
system of the microorganism, which reacts in the
presence of toxic metal.
Non -metabolism dependent: In this metal uptake is by
physico-chemical interaction between the metal and
the functional groups present on the microbial cell
surface.
 This is based on physical adsorption, ion exchange
and chemical sorption, which is not dependent on
the cells' metabolism. 
 Cell walls of microbial biomass, mainly composed of
polysaccharides, proteins and lipids have abundant
metal binding groups such as carboxyl, sulphate,
phosphate and amino groups.
 This type can be relatively rapid and reversible.
According to the location where the metal removed
from solution is found, biosorption can be classified
as……

 Extra cellular accumulation/ precipitation

 Cell surface sorption/ precipitation and
 Intracellular accumulation.

If toxic metals are present then the method may be

dependent on cells metabolism as the cells produce
certain compounds that favor the precipitation.
 Transport across cell wall or
membrane: 

• Heavy metal transport across microbial cell

membranes is mediated by the same mechanism
used to convey metabolically important ions such as
potassium, magnesium and sodium. 

• This method requires cell metabolism.

Physical adsorption:
• physical adsorption takes place with the help of van der
Waals' forces.
• biomasses of algae, fungi and yeasts ,through electrostatic
interactions between the metal ions in solutions and cell
walls of microbial cells can uptake the metals. 
Ion Exchange:
Cell wall of microorganisms contain polysaccharides, the
bivalent metal ions exchange with the counter ions of the
polysaccharides. 
For example, the alginates of marine algae occur as salts of K+, Na+, Ca 2+, and Mg2+.
These ions can exchange with counter ions such as CO 2+, Cu2+, Cd2+ and Zn2+
resulting in the biosorptive uptake of heavy metals
Complexation:
• The metal removal from solution may also take place
by complex formation on the cell surface after the
interaction between the metal and the active groups.
• Micro­organisms may also produce organic acids
(e.g., citric, oxalic, gluonic, fumaric, lactic and malic
acids), which chelate toxic metals resulting in the
formation of metallo-organic molecules.
• These organic acids help in the solubilisation of metal
compounds and their leaching from their surfaces.
• Metals may also be biosorbed or complexed by
carboxyl groups found in microbial polysaccharides
and other polymers.
Precipitation: 
• It may be either dependent on the cellular
metabolism or independent of it.
• In metabolism dependent precipitation, the biomass
react in the presence of a toxic metal producing
compounds, which favour the precipitation process.
• In the case of precipitation not dependent on the
cellular metabolism, it may be a consequence of the
chemical interaction between the metal and the cell
surface.

The various biosorption mechanisms can take place

simultaneously.
 Use of Recombinant bacteria for metal
removal:
• Metal removal by adsorbents from water and
wastewater is strongly influenced by physico-
chemical parameters such as ionic strength, pH
and the concentration of competing organic and
inorganic compounds. 

• Genetically engineered E.coli, which expresses

Hg2+ transport system and metallothionin (a
metal binding protein) was able to selectively
accumulate 8 mmole Hg2+/g cell dry weight.
Genetic engineering of bacteria to remove
toxic metals from the environment

E. coli bacterium
New gene/transport proteins

Hg2+-metallothein
Hg2+
Hg2+→Hgo
New gene/enzyme
 Desorption
• It is the regeneration of the biosorbant and
recovering the accumulated metals.

• This metal recovery process may be destructive

or non-destructive.

• Metabolism-independent biosorption is often

reversible and leads to non-destructive
desorption whereas metabolism-dependent
intracellular accumulation is often irreversible,
necessitating destructive recovery
• Regeneration of the biosorbent is done by washing
the metal- laden biosorbent with an appropriate
solution, the type and strength of this solution would
depend on the extent of binding of the deposited
metal. 

• Dilute solutions of mineral acids like hydrochloric

acid, sulphuric acid, acetic acid and nitric acid can be
used for metal desorption from the biomass.
• The desorption process should:
•  yield the metals in a concentrated form;
•  Restore the biosorbent to close to the
original condition for effective reuse
with undiminished metal uptake and
•  no physical changes or damage to the
biosorbent.
 LIMITATIONS
• Early saturation, i.e. once metal interacting sites
are occupied , uptake ceases and desorption is
required.

• The potential for biological process improvement

through genetic engg. is limited as cells are not
metabolizing.

• There is no potential for biologically altering the

metal valency state.
 Effect of Different Parameters :
• pH ---- At low pH the positively charged metal ions
are in competition with the protons in the soln. to
bind to the negatively charged surface of the active
site of bimass.
• So, at low pH metal uptake is less as cell wall remains
associated with H3O + ions, and the metals will face a
repulsive force.
• As the pH increases to 3-4 there’s a subsequent
increase of negative charge , so uptake increases.
• After pH 6 there is a drastic increase in metal uptake
due to formation of metal hydroxides with their
respective metal ions.
Adsorbent Mass:
• As the biomass increases the number of
binding sites for metals also increases.

• After some point sorption capacity reaches a

steady state or uptake decreases with
biomass conc. the active sites get blocked and
uptake decreases.
Contact Time:
• The rate of sorption is higher at the beginning
due to more biomass which provides
increased contact between metal and
biomass.
• As the active sites in the biomass are
exhausted uptake is controlled by the rate at
which adsorbate is transferred from exterior
to interior of adsorbant.
• Maximum removal time is within the first 45
min.. Equilibrium time is generally set as 4 hrs.
Initial Metal Ion concentration:
• Initial metal concentration provides an important
driving force to overcome all mass transfer
resistance of metal between aqueous and solid
phase.
• At higher conc. There is an aggregation of adsorbent
particles, such aggregation leads to a decrease in the
total surface area of adsorbent and increase in
diffusion path length.
• But after a certain conc. the uptake decreases due to
the blockage of active sites.
• The initial metal conc. Should be in the range of
200-600 ppm.
Effect of Temperature:
• Temperature effects a no. of factors for heavy metal
biosorption like :
i. Stability of initial metal ions in soln.
ii. Stability of micro organism-metal complex depending on
the site of biosorption.
iii.Effect on microorganism cell wall.
iv. The ionization of chemical moieties in the cell wall.
With increase in temp liquid viscosity decreases ,increasing
the adsorbate diffusion across the external boundary
layer.
It effects the equilibrium capacity of adsorbate depending
on whether process in exothermic or endothermic.
Sorption isotherm:
• This is a graphical representation showing a relation
between the amount adsorbed by a unit weight of
adsorbent and the amount of adsorbate remaining in
the test medium at equilibrium.
• The Langmuir and Freundlisch are the most common
models used for adsorption of metal.
• Langmuir eqn is based on some assumptions:
• All sites are equivalent
• Adsorption results in monomolecular layer of coverage.
• A molecule is adsorbed on a site independent of other
adsorbed molecules.
• Coverage is independent of binding energy and
• Constant temprature.
 Langmuir eqn.
The rate of attachment to the surface should be
proportional to the driving force times the area.
Driving force is the conc. In the fluid and
Area is amount of bare surface.
Affinity between biomass and diff metals is given by:

q = qmax b Ceq / 1+ b Ceq

Where, q is milligrams of metal accumulated per gram of the

biosorbent material;
Ceq is the equilibrium metal concentration in solution in mg/L;
qmax is the maximum specific uptake per unit mass of
adsorbent in mg/g and b is the Langmuir constant for ratio of
adsorption and desorption rates.
 Freundlisch eqn.

Linear form :
q = kF Ceq 1 /n

Where F and n are constant.

q is conc. Of metal ion adsorbed.
Ceq is equilibrium conc. Of metal ions .
These models can be applied at a constant pH.
These models are used in literature for modeling of
biosorption equilibrium in the presence of one
metal.