Physico-Chemical Characterization of A Biosurfactant Produced by
Physico-Chemical Characterization of A Biosurfactant Produced by
Physico-Chemical Characterization of A Biosurfactant Produced by
Pseudomonas Fluorescens
Surfactants
Surfactants are amphipathic molecules that reduce the surface tension between phases of differing polarity, such as oil and water.
Surfactants are used as emulsifiyng , dispersing, solubilizing, foaming agents. Nowadays surfactants are one of the most important substances for many fields of industry pharmacy, food industry, design of washing agents, petroleum industry, agriculture, environmental protection and remediation An excessive use of chemical surfactants leads to technogenic load on environment, flora and fauna, affects on food products
Biosurfactants
Types
Most microbial surfactants are complex molecules, comprising different structures that include peptides, glycolipids, glycopeptides, fatty acids and phospholipids.
Rhamnolipid Rhamnolipid
Surfactin lipopeptide
SOLUTION?
Various techniques are used for cell or biomass immobilized, such as flocculation, adsorption on surfaces, covalent bonding to carriers, cross linking of cells, encapsulation in a polymer gel, and entrapment in a polymeric matrix.
IMMOBILIZATION METHODS ADSORPTION ENCAPSULATION ADHESION ENTRAPMENT IN GEL OR POLYMERMATRIX
CELL IMMOBILIZATION!
Limits
Diffusional limitations Support plugging Product inhibition
Supports
Either natural biopolymers (polysaccharides, alginate, carrageenan, and agar) or synthetic polymer (polyacrylates, polyurethanes, and polyether's) can be used as gel-forming agents.
Calcium alginate
Calcium Alginate gel is widely employed for immobilization of whole cells since it is less toxic than synthetic polymers and easily gelled under mild conditions.
Entrapment in insoluble Ca alginate gel is recognized as a rapid, nontoxic, inexpensive, versalite and the most often used method for immobilization of cells.
Objectives
Study the feasibility and effect of immobilization of Pseudomonas fluorescens on the kinetics of biosurfactant production compared to free cells process. Biosurfactant Separation and characterization
Assays:
Surface tension (ST) (De Nouy Tensiometer Kruss K6), emulsification index (E24%), Biomass (dry weight) and pH
Immobilization Technique
Entrapment in calcium alginate
Optimum Conditions for immobilization were obtained by Response Surface Method with three factors : Sodium alginate concentration
Calcium chloride concentration Biomass loading To obtain maximum Responses of surface tension and E24
washing Sodium alginate
Cell suspension
Syringe(20G)
RESULTS
Biosurfactant production kinetics by free [A] and immobilized [B] cells of Pseudomonas fluorescens
75 60
)
10 8
) ) ) )
75 60 45
10
8
)
45 30 15 0 0 20 40 60
6 4
pH ( Biomass(
6 30 15 0 0
pH (
[A]
2 0 80 100 120
[B]
Time(hours)
Time(hours)
Biosurfactant production kinetics by free [A] and immobilized [B] cells of Pseudomonas fluorescens
Minimum ST (dyne/cm) Maximum E24 (%)
pH variations
Observations
Free cells
30
67
40-50
6.8-7.0
Immobilized cells
35
62
72
Biosurfactant separation
Easy recovery with acetone precipitation Yield (6g/l) Purer product with immobilized cell technique
Biosurfactant characterization
80
Surface tension (dyne cm )
-1
70 60 50 40 30 20 10
0 1 2 3 4 5 6 7 8 -1 Biosurfactant concentration(g L )
Wetting capacity
98,7
ability effect on hydrophobic surfaces Contact angle () (polystyrene Eau CH2I2 Formamide surface)coated with biosurfactant:
30,8 74,8
Good wetting
43
58
63,2
Solubilized Naphthalene(mg/l)
Solubilized Naphthalene(mg/l)
5% 10% 20% 0%
0 1 2 3 4 5 Boiosurfactant concentration(g/l)
1 2 3 4 5 Boiosurfactant concentration(g/l)
Conclusions
Diffusional limitations in alginate beads affected the
kinetic of biosurfactant production. Stable emulsions were obtained with immobilized cells. Easy separation Thermal and chemical stability increasing with pH Interesting characteristics for use in environmental and industrial applications at normal and extreme conditions.