Isolation of biosurfactant-producing and oil degrading

bacteria consortia
Suaad Hamad, Karina Law, Catherine Chen and Peter Morin Nissom*
Faculty of Engineering, Computing, and Science, Swinburne University of Technology (Sarawak Campus)
Jalan Simpang Tiga, 93350, Kuching, Sarawak, Malaysia
*Email: pmorin@swinburne.edu.my

INTRODUCTION
Bioremediation
Microorganisms that
can degrade oil are
used to remove
pollutants

Advantages

Biosurfactant producing bacteria

Natural process
Less expensive
Can remediate areas
that are not easily
accessible or inaccessible
to other technologies.

Sample collection

These bacteria have the ability to degrade oil

and utilise hydrocarbon.
They can be used in bioremediation and had
proven to give promising results.

Spot Fairy Cave

(undisturbed environment, may have
novel strains)

The crews are

trying to remove oil
spill by using
bioremediation.

Left : Pseudomonas
Right : Stenotrophomonas

METHODOLOGY
1.Screening for oil degrading & surfactant producing bacteria

2.

Dilution of soil samples and transfer to MSM

media with 1% oil

Identification of isolated strains

Dilution of soil samples and spread plate on NA

plate

Shake at 120rpm for 5 days and sub-cultured

for 6 times

Sub-culturing until single colony is obtained.

Perform gram staining method &

microbact kit test

Perform filter paper test & oil displacement

test for each sub-culturing

RESULTS & DISCUSSIONS

1.

Enrichment cultures (MSM + 1% oil)

2. Oil displacement test

3. The table of bacteria morphology

Graph of Oil Displacement Test

3.5

R = 0.9275
R = 0.8361

Diameter (cm)

3
2.5
2

sample
positive
negative

1.5
1

BACTERIA

SHAPE

GRAM

MORPHOLOGY

PLATE 1 ( 10-1)

ROD

MILKY WHITE
CONCAVE

PLATE 4 ( 10-4)

ROD

MILKY WHITE
CONCAVE

PLATE 5 ( 10-5)

COCCO
BACILLI

MILKY WHITE
FLUFFY

ROD IN CHAINS

MILKY WHITE
FLUFFY

ROD

NOT MILKY, CLEAR

SMALL

0.5
R = 0.9754

0
0

10

15

20

25

30

35

40

PLATE 6 ( 10-6)

Day

CONCLUSION
4.

In this project, bacteria were able to grow on the enriched media with
the hydrocarbon as sole energy source.
The bacteria with high potential of degrading oil & producing surfactant
were successfully being isolated & had identified on its genus.
Most isolates were from Stenotrophomonas genus. This genus might be
the novel bacteria that can degrade oil & produce surfactant derived from
the limestone sites.

FURTHER WORKS

ACKNOWLEDGEMENT

REFERENCES

Soil samples were stored in -80 for further

analysis.
Carry out PCR and Sequencing.
Identify 16S RNA of the potential bacteria.
Further analyze on bio augmentation test.
Test on other hydrocarbon (such as petroleum).

A very sincere thank you to Chua

Jia Ni, Rafika and Nurul for their
helping hand, guidance, advice and
moral support throughout the
preparation of this final year project.

Bioremediation n.d., Oppenheimer Biotechnology

Incorporated,
viewed
4
May
2012,
<http://www.slsc.org/uploadedFiles/Bioremediation.pdf>.
Margesin R. and Schinner F. (2001) Biodegradation and
bioremediation of hydrocarbons in extreme environments.
applied Microbiology and Biotechnology 56:650663.