1. BIOSURFACTANTS
SUBATHRA.S

2. BIOSURFACTANTS
Biosurfactants are proteins which present potential application in the
environment protection, organic substances treatments and oil recoveries.
Microbial surface active molecules able to reduce superficial and
interfacial tensions either in water solutions or in mixtures of hydrocarbons.
Low toxicity,
high biodegradability,
environment compatibility
and specific activity in extreme conditions.
when comparing with chemical surfactants.

3. Screening Organisms
Chroococcus minor BDU 20321
Synechococcus elongatus BDHKU 33801
Synechococcus elongatus BDU
Oscillatoria laetevirens BDU 10143
Oscillatoria formosa BDU40261
Oscillatoria willei BDU
Phormidium valderianum BDU 43131
Phormidium valderianum BDU140842
Phormidium tenue BDU 46241
Nostoc calcicola BDU 40302

4. CTAB TEST-Nostoc calcicola BDU 40302

5. SCRENNING METHODS 1 EMULSIFICATION INDEX(%)
1ml culture supernatant+4mlwater and 6ml of oil.
Mixed well
Leave it 24 hrs
Height of emulsion formed(cm)Х100
Total height(cm)
Organisms
Petrol
Castor oil
Crude oil
Gingelly oil
Neem oil
Diesel
Sunflower oil
Synechococcus elongatus BDHKU 33801
11.4 16
6.9 9
7.1
5.7
4.1
Synechococcus elongatus BDU 28 18 10
23.9
5.7
8.5 16
Nostoc calcicola BDU 40302
51.2 38
28.5 12
12.6
9.5
8.9

6. Emulsification Index(%)

7. Emulsification assay
3ml culture supernatant+1ml oil
Mixed well.
Leave it for 1hr
OD was measured at 400nm.

8. EMULSIFICATION ASSAY

9. Emulsification assay
1ml of Nostoc calcicola BDU culture supernatant .
7.5 ml of Tris- Mg and 1ml of oil was added.
Optical density was measured at 540nm.

10. Emulsification assay for Nostoc calcicola BDU40302

11. OIL SPREAD METHOD Petri plate filled with water and covered with oil.
Then supernatant was added.
Clear zone was formed on the oil.
Nostoc calcicola BDU was displaced the diesel.

12. Concentration of Biosurfactant
Extraction with Diethyl ether in the ratio of 1:3 volume.
The organic layer was taken and dried in the rotary vacuum.
PURIFICATON OF BIOSURFACTANT
Culture supernatant was taken and made up to pH 2 by using Conc. HCL and incubate for over night, precipitate was formed.
The precipitate was taken and carbohydrate test was done.

13. Nostoc calcicola Supplemented with Petrol Control 1% 5% 10% 25%

14. Nostoc calcicola Supplemented with Petrol Control 50% 75% 100%

15. Emulsification Index(%) for petrol supplemented Nostoc culture Concentration of oil Emulsification index(%)
Control
0.0 1% 4 5%
6.8
10% 11
25% 21
50% 45
75% 33
100%
0.6

16. Emulsification Index for Nostoc calcicola with petrol Control 1% 5% 10% 25%

17. Emulsification Index for Nostoc calcicola with petrol Control 50% 75% 100%

18. Nostoc calcicola supplemented with Petrol For seven days

19. Nostoc calcicola BDU 40302 Supplemented with Petrol for seven days

20. Growth Curve For Nostoc calcicola BDU 40302

21. Growth Curve For Nostoc calcicola BDU 40302

22. Nostoc calcicola Supplemented with Crude oil For seven days Control 1% 5% 10%

23. Nostoc calcicola Supplemented with Crude oil For seven days Control 25% 50% 75% 100%

24. Nostoc calcicola BDU 40302 supplemented with crude oil

25. Nostoc calcicola BDU 40302 supplemented with crude oil

26. Thin Layer Chromatography
To find –Carbohydrates, Lipids.
To Isolate DNA
Polymerase Chain reaction and
Sequencing.