50

REMOVAL OF CHROMUM, LEAD AND COPPER IONS FROM

INDUSTRIAL WASTE WATERS BY Staphylococcus saprophyticus

Semra Ilhan
1*
, Macit Nurba Nourbakhsh
2
, Serpil Kiliarslan
2
and Huseyin Ozdag
3

1
Department of Biology, Osmangazi University, 26480 Eskisehir, Turkey
2
Department of Chemical Engineering, Osmangazi University, 26480 Eskisehir, Turkey
3
Department of Mining Engineering, Osmangazi University, 26480 Eskisehir, Turkey

ABSTRACT
In this study, selective biosorption of chromium, lead and copper ions by
microorganisms from industrial wastewaters were investigated. Microorganisms were isolated
from soil and in this research a bacterium which was identified as Staphylococcus
saprophyticus was used. The effects of pH, temperature and initial concentration of metal ions
on the biosorption capacity were investigated. The optimum pH values for chromium, lead
and copper biosorption was found to be 2.0, 4.5 and 3.5 respectively. The maximum
adsorption was observed for Cr
6+,
Pb
2+
and Cu
2+,
at the initial concentrations of 193.66 mg
Cr
6+
/l; 100 mg Pb
2+
/l and 105 mg Cu
2+
/l and under these conditions the biosorption values
were found to be 88.66 mg Cr
6+
/l; 100 mg Pb
2+
/l and 44.94 mg Cu
2+
/l, respectively.
The results indicated that S. saprophyticus was suitable for biosorption of lead and
chromium from wastewaters.
Key words: Biosorption, adsorption, heavy metals, industrial wastewaters
Staphylococcus saprophyticus.
*Corresponding author. Tel.: +90 222 2290433/2419
fax: +90 222 2393578
E-mail address: silhan@ogu.edu.tr


INTRODUCTION
Metals are directly and/or indirectly involved in all aspects of microbial growth,
metabolism and differentiation. Many metals such as K, Na, Mg, Ca, Mn, Fe, Cu, Ni, Co, Zn,
Mo are essential for biological functions, whereas it is not known whether some others, such
as Al, Ag, Cd, Sn, Au, Sr, Hg, Tl and Pb have essential biological functions. All these
elements can interact with microbial cells and be accumulated as a result of physico-chemical
mechanisms and transport systems of varying specificity, independent on, or directly and
indirectly dependent on metabolism (Gadd, 1988). Some of these processes are of
biotechnological importance being relevant to metal removal and recovery from mineral
deposits and industrial effluents for industrial use or environmental bioremediation.
Turkish Electronic Journal of Biotechnology
Vol 2, p:50-57, 2004
Biotechnology Association

51
Metals are introduced into aquatic systems as a result of the weathering of soils and
rocks, from volcanic eruptions, and from a variety of human activities involving the mining,
processing, or use of metals and/or substances that contain metal contaminants (Laws, 1993).
The last group which has the highest potential of toxicity and pollution includes waste waters
coming from metal plating industry, automobile, electrical and electronic materials, home
appliances, pipes, caps, guns, mechanics and dye industries. Unlike other pollutants, heavy
metals are getting important because of the fact that they can not be decomposed by in situ
biological means. Virtually all metals, including the essential metal micronutrients, are toxic
to aquatic organisms as well as humans if exposure levels are sufficiently high (Laws, 1993).
It is, therefore, necessary to remove heavy metals from waste waters before discharge due to
effects of disturbing environmental quality and being harmful to human health.
The removal of metals from industrial effluents can be achieved by ion exchange,
chemical oxidation, chemical precipitation etc (Aksu et al., 1992). For advanced purification,
different physico-chemical methods such as active carbon adsorption, ion exchange and
reverse osmosis are used. As an alternative to these methods, recently, the method of the
removal of heavy metal contaminants by means of bacteria has been focused on. Biological
removal of heavy metal contaminants from aquatic effluents offers great potential when
metals are present in trace amounts (Fourest and Roux, 1992; Vinita and Radhanath, 1992).
Many microbial species such as bacteria, fungi, yeast and algae are known to be
capable of adsorbing heavy metals on their surface and/or accumulating within their structure
(Campbell and Martin, 1990; Luef, 1991; Mitani and Misic, 1991; Vinita and Radhanath,
1992;). It is possible that microorganisms can be used in the removal of toxic metal ions from
the water and even in the recovery of them by using these adsorption properties of the
microorganisms. Physical adsorption or ion exchange at the living or non-living cell surface is
very rapid and occur in a short time after microorganisms come into contact with heavy metal
ions. Accumulation occurs in living cells and is slow, related to metabolic activity (Gadd,
1990; Gadd, 1992, Nourbakhsh at al., 2002). Although there are a number of studies on
removal of heavy metals, the knowledge of the applications of biosorbents in the environment
and industry has not been clear yet.
In this study, some important parameters that should be considered in the removal of
heavy metals from the industrial waste waters were investigated and an approach concerning
with application to the industrial waste water containing heavy metals have been presented.
52

MATERIALS AND METHODS
Isolation and identification of microorganisms:
The bacterium strain used in this study was isolated from the soil samples undergone
enrichment process (Nourbakhsh at al., 2002). The enrichment was carried out in the shaken
culture of the medium with the following contents: 4.0 g/l D(+) glycose monohydrat, 0.8 g/l
KH
2
PO
4
, 4.0 g/l (NH
4
)
2
SO
4
, 0.1 g/l NaH
2
PO
4
, 0.2 g/l MgSO
4
.7H
2
O, 4.0 g/l yeast extract, 0.1
g/l CaSO
4
.H
2
O, 4.0 g/l pepton, 4.0 g/l nutrient broth, 0.1 g/l FeCl
3
.6H
2
O, 0.1 g/l
Na
2
MoO
4
.2H
2
O. Pure culture was obtained by inoculation to nutrient plates from the
enrichment culture. Isolates were investigated with respect to their removing capacity of lead,
copper and chromium from the solutions containing these heavy metal ions. The isolate
OGUB 003 was selected. Microscopic and biochemical tests were applied to this isolate
according to Bergeys Manual of Systematic Bacteriology and API 20E (Schleifer, 1986). The
genus to which the isolate belongs was determined.
Growth of Microorganisms and Biosorption
OGUB 003 was incubated at 27C and at 150 rpm for 40-48 hours in tripticase soy
broth (Oxoid). At the end of incubation, biomass was separated from medium by centrifuging
at 5000 rpm and it was kept in the oven at 50C to remove the free water as much as possible.
Then, it was suspended in deionized water in order to use it in the biosorption. 100 ml
solutions containing 100 mg/l Cr
6+
, 150 mg/l Pb
2+
, and 100 mg/l Cu
2+
were prepared from
stock solution containing 1 g/l Cr
6+
(K
2
Cr
2
O
7
), 1 g/l Pb
2+
(Pb(NO
3
)
2
), 1 g/l Cu
2+
(CuSO
4
).
Then, 4.0 g microorganism (gmo) were added to the medium (20 gmo/l) and adsorptions of
metals were investigated for different pH values adjusted by using HCl and NaOH at 27C.
The solution containing the biomass was agitated in a shaker of 150 rpm during the
adsorption. Samples taken at predetermined intervals were centrifuged and supernatants were
analyzed. The analyses of Cr
6+
, Pb
2+
, and Cu
2+
ions were carried out by atomic absorption
spectrophotometer (Perkin-Elmer) at 0.01 ppm sensitivity level after dilution of the samples.
By taking the determined optimum conditions into consideration, the capacity of
microorganism to remove the mentioned metals from an industrial waste water was searched
with the same method.

53
RESULT AND DISCUSSION
The isolate OGUB 003 was identified as Staphylococcus saprophyticus. First, for
biosorption of Cr
6+
, Pb
2+
and Cu
2+
ions by wet biomass of OGUB 003 optimum conditions
(pH, temperature, ion concentration) were determined. The highest biosorption of Cr
6+
was
determined at pH 2, temperature 27C, initial ion concentration 193.66 mg/l while for Pb
2+

this was realized at pH 4.5, temperature 27 C, initial ion concentration 100-150 mg/l. For the
Cu
2+
optimum adsorption conditions were determined to be pH 3.5, temperature 27C, 105
mg/l initial ion concentration. Obtained results are shown in Table 1, 2, 3 and 4.


54


Under the optimum conditions determined (Table 4) the highest uptake, %100 yield,
was determined for lead ions. Biosorption of the metals in their individual solutions by S.
saprophyticus was determined to be Pb
2+
>> Cr
6+
>Cu
2+
. The walls of Gram positive bacteria
are efficient metal chelators and in Bacillus subtilis, the carboxyl group of the glutamic acid
of peptidoglycan was the major site of metal deposition. Teichoic and teichuronic acids were
important binding sites in Bacillus licheniformis (Gadd, 1990). S. saprophyticus is a Gram
positive bacterium and it has similar cell wall properties as of other Gram-positive bacteria.
The study was focused on biosorption of lead ions due to the high recovery rate of lead
ions by S. saprophyticus. Because lead ions were fully biosorbed from the solution having a
100 mg/l Pb
2+
initial concentration, experiments were carried out on the solution of 250 mg/l
Pb
2+
initial concentration, in order to apply the process to waste waters containing higher lead
ion concentration. The equilibrium time was determined to be 2 hours because after 2 hours
the adsorption rate was insignificant. However it was observed that the uptake reached to 70%
during the first 15 min. This high uptake can be attributed to physical adsorption on the cell
wall consisting of peptidoglucan (Gadd, 1990). Figure 1 shows the metal ratio biosorbed.
55

The waste water samples taken from a plating unit in Eskiehir were used as industrial waste
water. The results of removal of Cr
6+
, Pb
2+
and Cu
2+
ions from an industrial waste water by S.
saprophyticus were given in Table 5.

While in the individual solutions the biosorption yield of the metals by S.
saprophyticus was 100%, 24,2% and 14,5% for Pb
2+
, Cr
6+
and Cu
2+
, respectively, in a mixed
solution these were 100 %, 25% and 24 % for Pb
2+
, Cr
6+
and Cu
2+
. Because the adsoption was
selective (Nakajima and Akaguchi, 1986) and high in the industrial waste waters having low
lead ion concentrations (0.17 and 0.47 mg/l Pb
2+
), it can be said that S. saprophyticus can be
56
used in tertiary treatment of the waste water containing lead ions at low concentrations (Vinita
and Radhanath, 1992).
As a result, it can be concluded that S. saprophyticus can be used in the removal or recovery
of heavy metal ions, especially lead ions from industrial waste waters. It can also be said that
the usage of S. saprophyticus can be successful to remove chromium element from the waste
waters containing higher levels of chromium ions. Further studies are needed to increase the
biosorption capacities of biomass and to develop appropriate technologies applicable in the
treatment of industrial waste waters.

Acknowledgments
Authors acknowledge the Research Foundation of Osmangazi University for support
the project as numbered 1997/15.

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