Biosorption of lead ,cadmium,and mercu

ry ions on loofa sponge immobilized bio

mass of Aspergillus terreus

指導老師 : 孫 逸 民 教授
學 生: 田 汶 玄

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 Introduction(1)
Heavy metals are discharged from
various industries such as
electroplating, metal finishing, textile,
storage batteries, mining, ceramic and
glass.

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 Introduction(2)
The commonly used procedures for
removing metal ions from wastewater
include chemical precipitation, ion
exchange, membrane separation,
reverse osmosis, and electrolysis.

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 Introduction(3)
However, these techniques have certain
disadvantages such as incomplete
removal, high reagent and energy
requirements, generation of toxic sludge.

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 Introduction(4)
The search for alternative treatment
techniques has focused attention on the
use of biological materials such as
algae, fungi, yeast and bacteria for the
removal and recovery technologies and
because of the better performance and
low cost of these biological materials .

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 Introduction(5)
The purpose of the present study is to pr
ovide an immobilized biosorption system
using a low cost, physically strong, and
highly porous immobilization matrix; loof
a sponge.

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 Introduction(6)
The study investigated the use of loofa s
ponge immobilized fungal biomass as a
biosorbent for the removal of lead, cadmi
um ,and mercury from aqueous solution.

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Materials and Methods
 Microorganism
The Aspergillus terreus was maintained by pu
re culturing on potato dextrose agar slants at
28 ℃ for 7 days.

Mycelium suspensions from 7-day old culture

s, preculture on glucose at 28 ℃ for 24 hours.

Then, above Mycelium suspensions was main

culture for 15 days.

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 culture medium
medium concentration (g/l)
KH2PO4 5
K2HPO4 5
FeSO4 ． 7H2O 0.2
MnSO4 ． 4H2O 0.1
ZnSO4 ． 7H2O 0.2
CuCl2 ． 2H2O 5
H3BO3 11
(NH4)6Mo7O24 ． 4H2O 5
Mono-hydrate sodium 12.5
glutamate
Glucose 45
MgSO ‧7H O
4 2
0.1
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 Pretreatment of loofa sponge

The fibrous sponge was cut into discs of

approximately 0.2g , soaked in boiling
water for 30 min, thoroughly washed
under tap water, and left for 24 h in
distilled water, changed 3-4 times.

The sponge piece were oven dried at 70

℃.

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Immobilization of A.terrues within lo
ofa sponge
The mycelium suspension (2.5 ml) was inoc
ulated into 250 ml flasks containing 50 ml of
the growth medium and pre-weighed loofa s
ponge discs.

The inoculated flasks were incubated at 28

℃ and shaken at 200 rpm.

After 15 days of incubation, loofa sponge im

mobilized A.terrues were harvested from the
medium, washed twice with distilled water a
nd stored at 70 ℃ until use.
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 Biomass of fungal growth

The dry weight of the fungal biomass ent

rapped within sponge pieces was determ
ined by weighing sponge pieces before a
nd after fungal growth after 48 h drying a
t 70 ℃.

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 Metal solutions

HgCl2 (Riedel-de Haën)

CdCl2‧2.5H2O (Panreac )
PbCl2 (J. T. Baker)

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 Biosorption studies
The biosorption capacity of FBILS (fungal biomass immobi
lized in loofa sponge) was determined by contacting 100
ml metal solution of known concentration in 250 ml flasks.

The metal solution was shaken on an orbital shaker at 200

rpm and 30 ℃.

FBILS were separated from the solution by decantation.

Residual metal concentration in the metal solutions by ce

ntrifugation at 3000 rpm for 5 min

Supernatant was analysed for residual metal ions concent

ration was determined using flame atomic absorption spe
ctrophotometer.
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Effect of contact time and pH on met
al ion biosorption
For the determination of rate of metal bioso
rption by FBILS, the supernatant was analy
sed for residual metal ions after the contact
period of 10, 20, 30, 60, 120, 240 and 360 mi
n.

The effect of pH on metal ions sorption by F

BILS was determined by equilibrating the s
orption at different pH values of 2, 3. 4, and
5.

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 Chemical and physical stability of
FBILS
For the purpose, FIBILS was shaken on
an orbital shaker at different rotation
speed for 2 days.

Similarly, soaking of FBILS in buffer of

various pH (2.0–12.0 for 20 days) was
shaken on an orbital shaker at 200 rpm
for 20 days had weight loss 20% of the
FBILS were observed during this period.

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 data analysis
q = V(Ci - Ceq)/M

q is the metal uptake (mg metal ions /g dry weight of fungal bio
mass entrapped within sponge)

V is the volume of metal solution (ml)

Ci is the initial concentration of metal ions in the solution (mg/l)

Ceq is the final concentration of metal ions in the solution

M is the dry weight of fungal biomass

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Results and discussion
Fungal biomass immobilized within l
oofa sponge
(a) (b)

(a) natural loofa sponge covered with A. terrues biomass

(b) artificial loofa sponge covered with A. terrues biomass

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Growth curve — Aspergillus terreus
immobilized loofa sponge
biomass of immobilize loofa

1
0.8
sponge(g)

0.6 natural
0.4 artificia
0.2 l
0
0 1 3 5 7 9 11 13 15 18 23 30

growth phase (day)

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 Effect of different phase on metal
uptake by FBILS

400
Pb adsorbed(mg/g)

300

200

100

0
0 3 7 15 30
diferent phase(days)

100 ml single metal solution (6000mg/l) was contacted with different

days of FBILS on orbital shaker at 200 rpm at 30 ℃ at pH3 for 30 min

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Time-course profiles for metal ion
biosorption by FBILS
 P b a d so rb e d (
350
300
250
200
150
100
50
0
0 10 20 30 60 120 240 360 480 960
C d a d s o r b e d ( m g /g ) H g a d s o rb e d (m g /g )

time(min)

50
40
30
20
10
0
0 10 20 30 60 120 180 240 300
time(min)

40

30

20

10

0
0 10 20 30 60 120
time(min)
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 effect of pH on metal uptake by
FBILS
m etal ion ads orbed(m g/g)

350
300
250 Pb
200
Cd
150
100 Hg
50
0
1 2 3 4 5 6 7
pH

Metal solution was contacted with 15 days of FBILS at optimal condition

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Effect of temperature on metal bioso
rption by FBILS

350
metal ion adsorbed(mg/g)

300
250
200 Pb
150 Cd
100
Hg
50
0
0 10 20 30 40 50 60
temperature(℃)

Metal solution was contacted with 15 days of FBILS at optimal condition

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Effect of rotation rate on metal bioso
rption by FBILS
m eta l io n a d s o r b e d ( m g /g )

350
300
250
Pb
200
Cd
150
Hg
100
50
0
0 50 100 150 200 250
rotation(rpm)

Metal solution was contacted with 15 days of FBILS at optimal condition

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Effect of initial metal ion
concentration
 P b ad s o rb ed (m g /g 400
300
200
100
0
0 1000 2000 3000 4000 5000 6000 7000 8000
H g a d s o r b e d ( m g /g )

initial concentration (mg/l)

50
40
30
20
10
0
C d a d s o rb e d (m g /g )

0 200 400 600 800 1000 1200

initial concentration (mg/l)

35
30
25
20
15
10
5
0
0 100 200 300 400 500 600

initial concentratiom (mg/l)

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 Chemical stability of LIBCS

100
80
weight loss (%)

60 before weight
40 after weight
20
0
2 4 6 8 10 12
pH

soaking of FBILS in buffer of various pH (2.0–12.0 for 20 days)

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 physical stability of FBILS
100
80
weight loss (%)

60 before weight
40 after weight
20
0
50 100 150 200 250
rotation speed(rpm)

soaking of FBILS in pH7 buffer for 2 days

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Conclusions
 Loofa sponge is an effective immobilization carrier for
the entrapment of A.terreus to produce FBILS.

The biosorption capacity of FBILS for single metal decr

eased in the order Pb > Hg > Cd

FBILS showed an excellent potential for the removal le

ad(II) from aqueous solution.

FBILS showed an good physical and chemical stability.

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Thanks for your attention