Isolation and Characterization of

Biosurfactant Producing Bacteria

and
Optimization of Biosurfactant
Production

Sunita Sanjay Dange

ADT’s
Sharddbai Pawar Mahila Mahavidyalaya
Shardanagar,Baramati

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 INTRODUCTION
• Biosurfactants
• Applications
• Significance wrt chemical
surfactants
• Action
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 INTRODUCTION
• complex molecules including peptides, fatty
acids, phospholipids, glycolipids, lipopeptides
• used industrially as adhesives, flocculating,
wetting and foaming agents, de-emulsifiers and
penetrants.
• Biodegradability, Biocompatibility and
digestibility, Availability of raw materials,
Specificity
• Environmental control: Processes for stabilization
of industrial emulsions, control of oil-spills,
biodegradation and detoxification of industrial
effluents and in bioremediation of contaminated
soil can be favored with use of biosurfactants.
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 AIM
Aim of the present study is to focus special
emphasis upon the screening of over-
producer microbial strains and optimization of
cultural parameters.

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 OBJECTIVES
•Isolation of Microorganisms in Bushnell Hass
broth
•Screening by using Blood Haemolysis Assay
•Identification
•Production of biosurfactants .
•Extraction and purification of biosurfactants
produced.

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 RESULTS

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 RESULTS
Colony Size Shape Colour Margin Elevation Opacity Consistancy
characterstics

5mm Circular Bluish- Entire Convex Opaque Smooth

green

Gram staining - Gram negative( short thin rods)

Motility - Motile

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 Sr no Biochemical test Result
1 Ala-Phe-pro-Arylamidase -
2 H2S production -
Beta-glucosidase BGLU-26 Lysine
3 -
D carboxilase LDC-
4 4 L – Proline-Arylamidase +
5 Saccharose/ Sucrose -
6 L –Lactate alklinization +
7 Glycine – Arylamidase -
8 O129R resistance +
9 Adonitol -
10 Beta- N-acytil-Glucosaminidase -
11 D- Maltose -
12 Lipase -
13 D-Tagatose -
14 Alfa- Glucosidase -
15 Ornithine D-Carboxylase -
16 Glu-Gly-Arg-Arylamidase -
17 L-Pyrrolydonyl-aryamidase -
18 Glutiamyl arylamidase pNA +
20 Palatinose -

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 Optimization of production by combination of different parameters

Parameters pH Temp. Inoculu Agitation Incubation Nacl

m size period
Conditions 7 30 0 C 1% 120 rpm 72 hrs 0.5%

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 Sr.no. Strain

Carbon source O.D E24

at 540 nm index

1 Olive Oil 0.64 48

2 Coconut Oil 0.30 39
3 Dextrose 0.11 35
4 Engine oil 0.42 50

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 Sr.No. Isolate

Carbon Diameter of
source zone (cm)

1 Olive Oil 5.2

2 Coconut Oil 7.9

3 Dextrose 1.0
4 Engine oil 6.2

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 Oil Displacement Tests

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Production of biosurfactant at optimized conditions:
by an isolate- Optimum conditions.

Temperature- 300 C
pH – 7
NaCl concentration -0.5%
Carbon source- Coconut oil
Nitrogen source-NH4NO3
Aeration-120rpm
Production medium-Nutrient broth with 1%glycerol

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 16s rRNA identification
Closest Pseudomonas aeruginosa
Neighbour

Strain JCM 5962(T)

Citation (Schroeter 1872) Migula

1900

Accession No. BAMA01000316

Pairwise 100
Similarity (%)

Diff/Total nt 0/1486
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 Stability of the product
•PH range-4-11
•Temperature range(0C)-20-50
•Salt Concentration-0.5-3.5%
•Shelf life-20days
•Toxicity(Fish Bioassay)- Low toxicity(above
8grams per litre)

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 Applications

•Soil Remediation-Use of soil columns

•Soil Wettability- From 5grams per litre

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•The latest discoveries in the biotechnological
and nanotechnological fields are applicable to
biosurfactants also. Further research on the use
of the produced biosurfactants as a “green”
alternative for high-performance nanomaterials
production is possible.
Prerequisites for making biosurfactant
production more profitable and economically
feasible include optimized growth/production
conditions and novel and efficient multi-step
downstream processing.
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