ISOLATION AND CHARACTERIZATION OF

GLYCOLIPID BIOSURFACTANT PRODUCED BY

Pseudomonas otitidis STRAIN ISOLATED FROM
CHIRIMIRI COAL MINES, INDIA

Afifah Nur Aini (176090100011006) Microbial Physiology and

Master Program in Biology Biochemistry Class
Brawijaya University April 4, 2018
 Soil sample was Methods
isolated from coal
mining site Selected bacteria: P4

Grown in Carbon Free Characterization

Mineral Media (CFMM)
supplemented with Phenotypic Phylogenetic Physiological
pyrene (colony morfology) 16S rRNA & biochemical
tests
Incubated at 30oC
with shaking at
150 rpm for 7 days Biosurfactant production

Biosurfactant
Positive pyrene Effect of different
stability in different
degrading bacteria C and N sources
parameters
show an increase
in the turbidity
Production of Biosurfactant in P. otitidis P4

BS is produced from
the late exponential
phase until the end of
the stationary phase
 Biochemical Characteristics of P. otitidis P4 – Carbon Source

7 Biomass Yield BS Yield

Coal, oil,
Concentration (g/L)

6 gasoline
5
PAH
4
(polycyclic
3 aromatic
2 hydrocarbons)
1
Pyrene (most
0
abundant)

It is only natural if biosurfactant-producing

bacteria can
Carbon utilize pyrene.
source
 BIOSYNTHESIS
PATHWAY OF
BIOSURFACTANT
IN Pseudomonas
Biochemical Characteristics of P. otitidis P4 – Nitrogen Source

Nitrogen
depleted

Reduced activity
The production of
Catalyzes the
of isocitrate oxidation of isocitrate biosurfactant
dehydrogenase to 2-oxoglutarate enhanced when
grown in nitrogen
Accumulation of limiting
isocitrate in
mitochondria
condition.

Isocitrate Isocitrate
Biosynthesis Biosynthesis of
transported to converted to
of fatty acids biosurfactant
cytosol acetyl-CoA
The Stability of Biosurfactant

• BS is stable at high
temp. (80-100oC),
potential to apply in
various industry
• Highest emulsification
index at 40oC, pH 7
and 40% NaCl
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