International Journal of Civil & Environmental Engineering IJCEE Vol: 10 No: 1
Application of Biosurfactant Produced by Azotobacter vinelandii AV01 for
Enhanced Oil Recovery and Biodegradation of Oil Sludge
Qomarudin Helmy, Edwan Kardena, Zeily Nurachman and Wisjnuprapto
Abstract - The problem of petroleum waste management
is giving a due consideration of the national level. Large
quantity of dehydrated oil sludge, generated in the
disposal process of oil-containing sewage in Indonesia
that needs to be rendered harmless to human and to the
environment. Microbial degradation has been accepted as
an important method for the treatment of oil sludge by
employing indigenous or extraneous microbial flora. The
purpose of this study was to investigate the performance
of biosurfactant in its attempt to recover oil from oil
sludge and in enhanced biodegradation of oil sludge
process. Measurement of biosurfactant production
indicated that the maximum production occurred at the
end of exponential growth phase (48h). The
emulsification capacity of the biosurfactant also stables
under thermal and ionic strength treatment that meet any
requirement for application in the oil recovery and
degradation. In the oil recovery preliminary test,
biosurfactant have the capability to recover oil up to 15%
from oil sludge. For oil sludge biodegradation assay, it
was found that addition of petrofilic consortia increased
the removal efficiency up to 55%, while addition of
biosurfactant in this reactor increased the total efficiency
of 70% after 70 days of incubation. These results suggest
that both petrofilic consortia and biosurfactant addition
stimulate the biodegradation and overcome the limitation
of petroleum hydrocarbon degradation process.
Keywords: A. vinelandii, biosurfactants, biodegradation,
oil sludge, oil recovery, crude oil
1. INTRODUCTION
Oil, being an essential energy source, is both the
lifeblood and a liability of many industrialized
nations. The use of crude oil as an energy source has
allowed many nations to develop a high standard of
living. Continued economic growth will increase the
demand for oil, which must be met by current
production technologies or by new discoveries.
Traditional oil recovery technologies under the
umbrella of chemically enhanced oil recovery
(CEOR) can recover a maximum of 40–45% of the
oil initially in place, in two stages, namely, primary
and secondary recovery [1, 2].
Manuscript received Januari 9, 2010. This work was supported in
part by ITB Research Grant and Directorate general of higher education
(DIKTI), Government of Indonesia for the project grant
No:324/SP2H/DP2M/III/ 2008 in environmental biotechnology.
Q. Helmy, E. Kardena and Wisjnuprapto is member of the Water
and Wastewater Engineering Research Group, Faculty of Civil and
Environmental Engineering, Institut Teknologi Bandung (ITB)
Indonesia. Ganesha 10 Bandung-West Java. Tel. No. +62 817 023 5878,
helmy@tl.itb.ac.id; kardena@pusat.itb.ac.id; wisjnu@bdg.centrin.net.id.
Z. Nurachman is member of the Biochemistry Research Group,
Faculty of Mathematics and Natural Sciences in the same Institute,
zeily@chem.itb.ac.id
One of the most encountered pollutants in oil
production companies is the formation of oil sludge
that is entrapped with the effluents during treatment
and conditioning of the wells produced crude oil
through treatment process facilities. Most of the oil
sludge is piled up outdoor without any treatment,
and poses a serious environmental problem. The
hydrocarbons in the sludge penetrate from the top
soil into the subsoil slowly, presenting a direct risk
of contamination to subsoil and groundwater [3-5].
On the other hand, the light hydrocarbons in the oil
sludge vaporize, leaving behind a layer of oil-
containing dust of soil which blows upwards to
pollute the air. Therefore, the oil sludge should be
treated to prevent harm to environment. Although
burning of the sludge may be simple and easily
adaptable, this technique has undesirable hazard in
air pollution. Cleaner technologies are needed due to
the environmental friendly such as microbial
degradation concept [6-9].
The oil sludge is attributed to two major factors
controlling in its formation. First is the inorganic
residue consisting of sediments, sands, scales, dust
and the second is the precipitation of paraffinic wax.
Since wax precipitates are sparingly soluble in crude
oil, temperature changes are the reason behind wax
precipitation. In addition to the above reasons, the
oxidation of heavy organic material in crude oil due
to climate changes or from oxidizing
microorganisms and also the interruption in material
balance due to losses of volatile components and the
tendency of asphaltene, resin and polymeric
compounds to precipitate all are the reasons of oil
sludge formation.
As the oil price rised during year 2008, attempt
to recover oil from oil sludge became advantageous
both from economic and environmentally point of
view. Production of oil sludge as oil refinery by-
product in Indonesia exceeding 2,000 ton.day-1. This
huge oil sludge production refer to environmental
problem because of its characteristics and handling.
Many technologies are employed to clean up
contaminated sites including various chemical and
physical methods such as excavation, thermal
evaporation and soil vapor extraction.
Bioremediation has been accepted as an important
method for the treatment of oil sludge by employing
indigenous or extraneous microbial flora. Under
certain conditions, living microorganisms primarily
bacteria can metabolize various classes of
hydrocarbons compound [10]. Since hydrocarbons
International Journal of Civil & Environmental Engineering IJCEE Vol: 10 No: 1
contain high organic matter, it is not surprisingly if it
can be used as a carbon source by microorganism.
The microorganisms in this process are effective in
oxidizing the dissolve organic compounds, including
some emulsified oil [11]. One of the important
factors in oil biodegradation rate is its solubility.
This related to the bioavailability of contaminant to
the microbial attack. One promising approach to
increasing the biodegradation rates of organic
compounds with limited water solubility is the
addition of biosurfactant [12, 13].
In this paper, we report the possible application
of biosurfactant produced from A. vinelandii in the
oil industry, investigated the performance of
biosurfactant in its attempt to recover oil from oil
sludge and in enhanced biodegradation of oil sludge
process.
2. MATERIAL AND METHODS
2.1. Reagents.
All chemicals were of reagent grade, purchased
from Merck, J.T. Baker and Sigma Chem Co.
Growth media were purchased from Oxoid ltd.
2.2. Crude Oil and Oil Sludge.
Crude oil and oil sludge samples was obtained
from Duri Oil Field Pekanbaru and Balongan Oil
Field Indramayu Indonesia, respectively.
2.3. Bacterial strain and Culture Conditions.
Azotobacter vinelandii AV01 was used in
producing biosurfactant, while the petrofilic
consortia containing Bacillus cereus BL01,
Pseudomonas stuzeri BL02, Acinetobacter sp.
BL03 and Bacillus sp BL04 were used in the
biodegradation assay of oil sludge. All bacteria were
obtained from the Culture Collection of
Environmental Biotechnology Laboratory-
Environmental Engineering Department, Institute
Technology of Bandung, Indonesia. A. vinelandii
was maintained at 4°C on mannitol enrichment agar
slants containing (l-1): 20 g mannitol, 20 g yeast
extract, 20 g tryptone, and 15 g of agar. While each
petrofilic bacteria was maintained at 4°C on Nutrient
Agar covering with 1 drop of crude oil. Sub-cultures
were made to fresh agar slants every 1 month to
maintain viability.
2.4. Biosurfactant Production.
Cultures of A. vinelandii were grown on a
minimal basal medium (MB) which composed the
following components (l-1) of distilled water: 1.5 g
of K2HPO4; 0.5 g of KH2PO4; 0.2 g of MgSO4; 0.25
g of (NH4)2 SO4; and 20 g glucose as substrate. 10
ml Trace Element solution was added per liter of
MB medium. The compositions of this trace element
(l-1) are 12 g of Na2EDTA2.H2O; 1 g of CaCl2; 0.4 g
of ZnSO4.7H2O; 10 g of NaSO4; 0.4 g of
MnSO4.4H2O; 0.1 g of CuSO4.5H2O; 0.5 g of
Na2MoO4.2H2O [14]. The medium was sterilized by
autoclaving at 121°C for 15 min. The inoculums of
A. vinelandii was prepared by transferring cells
grown on a slant to 250 ml Erlenmeyer flasks
containing 50 ml of MB broth. Culture was
incubated in an orbital shaker at room temperature,
110 rpm for 2 days. The MB containing 106 cells/ml
was used to initiate growth using 2% (v/v)
inoculums. Biosurfactant production was carried out
in a 10 liter capacity of fermentor at 37oC with
agitation speed of 100 rpm and aeration rate of 0.176
ft3.min-1 for 2 days.
2.5. Crude Biosurfactant Isolation
The fermentation broth was centrifuged at
13.000 rpm for 30 minute to obtain a cell free broth.
After centrifugation, the supernatant was then
dissolved in a 4 N hydrochloric solution and allowed
to stand overnight at 4°C, followed by the
biosurfactant extraction step with a chloroform
solvent at room temperature [15]. The organic layer
was transferred to a round-bottom flask and the
aqueous layer was re-extracted two times for
complete recovery of biosurfactant. The organic
phases were combined yielding a viscous brown-
colored crude biosurfactant product and then
evaporated to remove the solvent; the residue was
collected and weighted. Vermani [16] method was
used to determine the exopolysaccharide fraction of
biosurfactant. A mixture of 1:2 (v/v) biosurfactant
and chilled acetone were agitated and stand
overnight to precipitate. Formed precipitate were
filtered and gravimetrically analyzed.
2.6. Emulsification Index (E24).
To determine the emulsification index, Batista
[17] method was applied. Centrifugation at 13,000
rpm to separate biosurfactant from microorganism
cells yielding a cell free broth. A mixture of 1:1
between biosurfactant and crude oil is agitated for
about 2 minute then stabilized for 24 hour.
Emulsification index (%) determined by measuring
the column height of emulsified oil against its total
height multiplied by 100 times.
2.7. Emulsification Stability Test
Stability studies as described by [18], were done
using the cell free broth obtained by centrifugation
process as mentioned above.
Effect of extreme temperature. Five milliliter of cell
free broth was exposure with various temperature
conditions: 4oC, 27oC and 120oC for 30 min and
allowed to cool to room temperature, then the
emulsification index were measured using described
protocol.
International Journal of Civil & Environmental Engineering IJCEE Vol: 10 No: 1

Effect of extreme ionic strength. To study the pH
stability of biosurfactant, five milliliter of cell free
broth were exposure with various pH environments:
pH value between 2-12.
2.8. Oil Recovery from Oil Sludge
Adapting the soil washing method from [19]
with some modification, 1:1 ratio of oil sludge and
biosurfactant poured into 10 ml centrifuge test tubes.
The test tube was then shaken with vortex for 5
minute and allows settling. After that the test tube
was centrifuged at 6,000 rpm for 15 minute yielding
3 phases of oil emulsion, remaining oil sludge
matrix and liquid solution. Oil emulsion recovered
was then measured in volume unit.
2.9. Total Petroleum Hydrocarbon (TPH)
Measurements.
Measurement of TPH was conducted with
gravimetric method as described by [20]. Sample
was extracted with n-hexane, the organic layer were
pooled and dried by evaporation of solvents. After
evaporation, the amount of residual TPH recovered
was weighted.
2.10. Biodegradation Assay
To determine the performance of petrofilic bacteria
in degrading oil sludge, a preliminary
biodegradation assay developed and set up as
follows:
• Control-l: without addition of both petrofilic
inoculums/P and biosurfactant/B.
• Control-2: without P; add 2% (v/v) B.
• Reactor-1: add 2% (v/v) P; without B.
• Reactor-2: add 2 % (v/v) each of P and B.
• Oil sludge initial concentration was 100.000
ppm of TPH and incubation time for
biodegradation assay was 70 days.
TPH concentration and growth of petrofilic
bacteria were observed on certain time.
3. RESULT AND DISCUSSION
3.1 Biosurfactant Production
The reason we choose A. vinelandii are first, A.
vinelandii is a rhizosphere bacterial strain that able
to produce biosurfactant constitutively from water
soluble substrate without any addition of inducer
substrate; second, A. vinelandii known have the
ability to utilized free nitrogen from the air; and
third, the ability to produce biosurfactant from water
soluble substrates is preferred because single-phase
fermentation is simpler than biphasic fermentation
that usually occurred when hydrocarbon based
substrates were applied. Growth and biosurfactant
production from A. vinelandii with 2% glucose as
sole carbon source was described in the Figure 1.

1.6 16 100
Biosurfactant production (g/L)

1.4 14 90

Emulsification Index (%)

80
1.2 12
70
Biomass (g/L)

1 10 60
0.8 8 50

0.6 6 40
30
0.4 4
20
0.2 2 10
0 0 0
0 20 40 60 80 100
Incubation Time (hour)

Fig 1. Growth (solid circle symbol), biosurfactant production (open circle symbol) and emulsification
activity (solid square symbol) profiles of A. vinelandii grown in fermentor with 2% (v/v) glucose as a carbon
source at 37oC.

The biosurfactant production started to increase
during the exponential phase, reaching its maximum
after about 48 h (9.81 g/l). These results indicate that
the maximum of biosurfactant biosynthesis from
glucose occurred predominantly at the end of the
exponential growth phase. The emulsification
activity of the cell free broth increased up to 90% in
the first 24 hour of incubation, whereas surfactant
accumulation increased during this period and start
to decrease after reaching its maximum synthesis.
This might be due to biosurfactant were used as
carbon source by A. vinelandii. Similar result
International Journal of Civil & Environmental Engineering IJCEE Vol: 10 No: 1

reported by [18], that grown C.lipolytica with 10% effectiveness of biosurfactant produced from A.
canola oil and 10% glucose. Biosurfactant vinelandii was pH stable at pH range 2-8.
concentration reaching its maximum production after A
48h at the end of exponential phase and start to 100
decrease in a longer incubation time. While [21],
used the molasses and corn steep liquor as the 80
primary carbon and nitrogen source to produce

E24  (%)
rhamnolipid biosurfactant from Pseudomonas 60
aeruginosa GS3. Maximum surfactant production
occurred after 96h of incubation, when cells reached 40
the stationary phase of growth.
20
3.2. Emulsification Stability Test: effect of
0
temperature and pH
Interest in biosurfactant is growing due to their 4 27 120
applications in the protection of the environment and Temperature (oC)
in the oil industry. Their environmental uses are
principally to the bioremediation of petroleum B
hydrocarbon contaminated sites [22]. In the oil 100
industry, they are used in microbial enhanced oil
recovery (MEOR) and or biosurfactant-mediated 90
enhanced oil recovery, facilitate transportation of
heavy crude oil through pipelines and in the cleaning 80
E24  (%)
of contaminated vessels [1]. Application of
biosurfactant in enhanced oil recovery process must 70
meet any requirement involving the extreme
condition of oil reservoir [23]. These processes 60
frequently involve exposure to extremes of
temperatures, pressure, salinity, pH and organic 50
solvents. Stability studies were done using cell-free 2 6 8 12
broth obtained by centrifugation the culture at pH
13.000 rpm for 30 minute as described in methods.
The emulsification index of the culture broth
free of cells was stable when stored at 4oC either at Fig 2 Effect of temperature (a) and pH (b) on the
room temperature (27o C). It is interesting to observe emulsifying activity of cell free broth of A.
that the emulsification capacity of the biosurfactant vinelandii grown in fermentor with 2% (v/v) glucose
remain stable after heating for 30 minutes up to 1200 as a carbon source at 37oC.
C. The effect of thermal treatment (chilled/heated)
on the activity of the biosurfactant from A. 3.3. Oil recovery from oil sludge
vinelandii cultivated in minimum basal medium with For many countries like Indonesia, domestic oil
2% glucose as carbon source showed that no production is in decline and the likelihood of
appreciable changes in emulsification capacity discovering neither large nor new oil reserves is low.
occurred. Similar result reported by [18] by growing These countries must then rely on foreign imports,
C. lipolytica on canola oil and glucose to produce which can slow economic growth and employment
biosurfactant that reach its maximum emulsification and aggravate trade deficits. As the oil price rised
activity at high temperature (120oC). In contrary, during year 2008, attempt to recover oil from oil
liposan from C. lipolytica was found to be relatively sludge became advantageous both from economic
stable between 30oC and 90oC, but lost almost 60% and environmentally point of view. Production of oil
of its activity after boiling for 1 h [24]. The effect of sludge as oil refinery by-product in Indonesia
extreme temperature and pH on the emulsification exceeding 2,000 ton.day-1. Economically attractive
activity of the culture broth free of cell can be seen because approximately 15% (see Table 1) of oil
in Figure 2. Extreme of pH could possibly transform could be recovered from oil sludge. Also
less surface-active species into more active environmentally attractive because the remaining oil
emulsifiers by denaturation of proteinaceous sludge became less in the oil content thus make it
components or by increased ionization. The easier to handle with. However, the low oil prices
that prevailed during the end of year 2008 lead to a
International Journal of Civil & Environmental Engineering IJCEE Vol: 10 No: 1

shift in focus from enhanced oil recovery to

enhanced oil remediation processes. Table 1 showed 3.4. Biosurfactant Enhanced Biodegradation of
the properties of oil sludge and the experiments Oil Sludge
parameter. The most successful attempt in using Biosurfactant are also used in emerging
biosurfactant to recover oil from oil sludge reported technologies like microbial remediation of
by [25]. The clean-up process was successful in hydrocarbon and crude oil–contaminated soils.
removing the sludge from the tank bottom, and it Hydrocarbon contaminants are removed from the
also allowed the recovery of more than 90% of the environment, primarily as a result of their
hydrocarbon trapped in the sludge. The recovered biodegradation, which is performed by native
hydrocarbon had excellent properties and could be microbial populations. Such biodegradation is
sold after being blended with fresh crude. The known to be time-consuming and new technologies
recovered 774 m3 (≈5550 barrels, at US $60/barrel) have been developed; for example the addition of
of oil is worth $333,000. biosurfactant help to stimulate the indigenous
microbial population to degrade hydrocarbons at
TABLE 1 rates higher than those which could be achieved
Properties of oil sludge and biosurfactant enhanced through addition of nutrients alone. Biosurfactant is
oil recovery from oil sludge experiment parameters. a well known surface active agent that generally
used in improving the viability of contaminant to the
Parameter Value
microbial attack.
Oil sludge properties The biosurfactant affect the biodegradation
Oil sludge density at 25oC (kg/m3) 867.8 process by increasing the solubility and dispersion of
Viscosity at 25oC (centipoises) 450 the compound [26]. There are two ways in which
Oil content (%) 30-33 biosurfactant affect which is increasing the surface
Experiment parameters area of hydrophobic water insoluble substrate.
Volume of oil sludge (ml) 5 Secondly is increasing the bioavailability of
Volume of biosurfactant (ml) 5 hydrophobic water-insoluble substances. A
Conc. of biosurfactant (g/l) 9.81 laboratory scale of biosurfactant enhanced
Shaking time (min) 5 biodegradation of oil sludge was conducted. Effects
Centrifuge speed (rpm) 6000 of addition of biosurfactant from A. vinelandii in the
Centrifuge time (min) 15 biodegradation process were shown in Table 2. We
Oil recovered (ml)1 0.75 used crude biosurfactant in the form of cell free
Oil recovered (%)2 15 broth directly without purifying the biosurfactant
1
means values from triplicate measurement. first for the simplicity reason of the experiments.
2
oil in emulsion

TABLE 2
TPH removal efficiency of oil sludge biodegradation in batch reactor. C1/Control 1 without addition both
Petrofilic inoculums/P and Biosurfactant/B; C2/Control 2 (–P, +B); R1/Reactor 1 (+P, –B); R2/Reactor 2
(+P, +B).
Biodegradation system TPH Removal Efficiency (%)1 Increased Removal Efficiency (%)
Oil sludge biodegradation
C1 12.4 6.4 (C1-C2)
C2 18.8 55.6 (C1-R1)
R1 68.0 20.9 (R1-R2)
R2 88.9 70.1 (C2-R2)
1
means values from triplicate measurement.

The low water-solubility of many hydrocarbons availability to biodegrading microorganisms [27].

reduces their availability to microorganisms and
limits the biodegradation process. It has been
assumed that biosurfactant can be used to enhance
the bioavailability of hydrophobic compounds. On
the other hand this low water-solubility increases
sorption of compound to surface and limits their
Once again, biosurfactant can enhance growth on
bound substrates by desorbing them from surfaces or
by increasing their apparent water solubility. Figure
3 showed the microbial growth and TPH profile of
control reactor. It was noticed that changes in oil
sludge environmental condition from its originally
International Journal of Civil & Environmental Engineering IJCEE Vol: 10 No: 1

slurry phase into more aqueous phase in the reactor,
triggering the indigenous bacteria in it to grow. For
the total plate count measurement, the CFU values
increased from 103.2 (CFU/ml) at day 0 and reach its
maximum to 105.4 (CFU/ml) in the first week of
incubation. Similar pattern occurred in the control
reactor-2 (without addition of petrofilic inoculants,
added by 2% v/v of biosurfactant only).
Biosurfactant addition make the oil sludge become
more soluble in the reactor, this shown by increase
in the microbial growth from 103.2 (CFU/ml) at day
0 and reach its maximum to 106 (CFU/ml) in the
first week of incubation. However, the degradation
process of oil sludge by mean of indigenous bacteria
predicted small enough/neglect able throughout the
experiment. TPH losses in control reactors mainly
due to weathering/physical influences [28, 29] such
as temperature shift, shaking condition,
volatilization of low molecular weight of
hydrocarbon [30] and photo-oxidation [31]. This
phenomenon confirmed the results reported by [32],
who observed disappearance of diesel compound in
the absence of biological activity reached 11% after
45 days of incubation. While addition of synthetic
surfactant and biosurfactant in the reactor increased
the oil sludge degradation rate by indigenous
bacteria to 12 and 21% after 60 days incubation,
respectively [33].

1.2E+04 8.0
7.5
1.0E+04
7.0
TPH of oil sludge  (ppm)

6.5
8.0E+03

Log  CFU  ml‐1
6.0
6.0E+03 5.5
5.0
4.0E+03
4.5
4.0
2.0E+03
3.5
C1 C2 BC1 BC2
0.0E+00 3.0
0 7 14 21 28 35 42 49 56 63 70
Time (day)
Fig 3. The indigenous microbial growth (open symbol) and TPH degradation (solid symbol) profiles in
batch control reactor system with 0 (circle/C-1) and 2 % v/v (square/C-2) of biosurfactant addition at 27oC.

Figure 4 shows that after 70 days of incubation,
a significant reduction of TPH (68%) occurred in the
biodegradation system supplemented with petrofilic
consortia/R-1. This positive result suggests that bio-
augmented bacteria could degrade TPH
significantly. Bioaugmentation also can be used to
increase the biodegradative capabilities of the
indigenous microbial population. Compared with
control reactor/C1, addition of petrofilic consortia
increased the removal efficiency up to 55%. Non
biological degradation (physical transformation) also
occurred in the process; however the biological
transformation dominated the process based on the
growth of bacteria observed during the process. For
the total plate count measurement, the CFU values
increased from 106 (CFU/ml) at day 0 to 107.5 and
106.7 (CFU/ml) in the first week and day 70
respectively. The presence of biosurfactant in
biodegradation system (R-2) increased the removal
efficiency up to 20% compared to those without
addition of biosurfactant/R-1. The present of
biosurfactant also increased the microbial growth
from 106.5 (CFU/ml) at day 0 to 108.8 (CFU/ml) in
the first week of incubation and 108.1 (CFU/ml) at
day 70. Similar result by [34], that examined the
effect of rhamnolipid biosurfactant to diesel/water
degradation from 0 to 80 mg/l significantly increases
biomass growth and diesel biodegradation
percentage from 1000 to 2500 mg VSS/l and 40-
100%, respectively.
International Journal of Civil & Environmental Engineering IJCEE Vol: 10 No: 1

1.2E+04 9.0

1.0E+04 8.5

TPH of oil sludge  (ppm) 8.0E+03 8.0

Log  CFU  ml‐1
6.0E+03 7.5

4.0E+03 7.0

2.0E+03 6.5
R1 R2 BR1 BR2
0.0E+00 6.0
0 7 14 21 28 35 42 49 56 63 70
Time (day)
Fig 4 The petrofilic consortia growth (open symbol) and TPH degradation (solid symbol) profiles in batch
reactor system with 0 (circle/R-1) and 2 % v/v (square/R-2) of biosurfactant addition at 27oC.

Our findings show that the addition of both
petrofilic consortia and biosurfactant favors the
biodegradation of the oil sludge. The limiting
condition in the degradation of hydrocarbon and
other PAH is their insolubility, thus decreasing the
efficiency and rate of degradation. This limitation
can be overcome either by addition of surface-active
compounds surfactant to the growing culture, thus
making hydrocarbons more water-soluble and
available for the cell to degrade, or by production of
its own surfactant by the augmented organisms to
facilitate uptake. The presence of biosurfactant also
stimulate the catabolism of hydrocarbon by mean of
co-metabolism process since biosurfactant are
organic compound and readily degradable to
microorganism.
4. CONCLUSION
Referring to the Indonesian Ministry of
Environmental (regulation no. 128/2003), it was
mandatory to manage a safe treatment of these
wastes (oil sludge) and also disposed off in an
environmental friendly manner. It is regulated that
the final concentration of TPH must less than 10.000
mg/kg. Based on the data presented in this paper
indicate that the petrofilic consortia have the
capability to degrade oil sludge to comply with the
regulation above.
ACKNOWLEDGMENT
Q. Helmy thank S. Hidayat for contributing in part
of the research. The authors also gratefully
acknowledge Prof. Naoyuki Funamizu, Laboratory
of Engineering on Sustainable Sanitation, Hokkaido
University for facilitating QH in the research
fellowship program in Japan.
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Q. Helmy is a researcher in Institut Teknologi Bandung
(ITB), Indonesia. He received his BSc in Biology (2001),
Master in Environmental Engineering (2006). Currently,
he is a Doctorate Student at ITB. His research areas
include environmental biotechnology. He is a member of
International Water Association (IWA) and the
Indonesian Society for Microbiology, and received
research sandwich program from Hokkaido University,
Japan.