Vol. 1(2) SEP 2011 - SAVAP International

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Academic Research International ISSN: 2223-9553 Volume 1, Issue 2, September 2011 RNase (1ug/ml) - - + + + + - - - - - - - - Lysozyme (10ug/ml) - - + + + + - - - - + + + + Pepsin (10ug/ml) - - - - - - - - - - - - - - + Sign shows the appearance of zone of inhibition, - Sign shows the absence of zone of inhibition Supernatant of the bacterial strains cultures were heated and it was observed that heat destroyed the antibacterial activity of all the five selected bacterial strains. Which showed that the compound was heat sensitive (Table 3). For the chemical analysis T.L.C was done and for this purpose bacterial strain was grown on the nutrient agar having lawn of sensitive strain and after 24 hours the piece of gel from the zone of inhibition was dissolved in the diethyl ether and then this diethyl ether was allowed to evaporate and this was done twice and a thick mixture was prepared, which was yellow in color. T.L.C. or thin layer chromatography is similar to paper chromatography, but instead of a paper, an aluminum sheet coated with a thin layer of an absorbent material, such as finely powdered silica gel or aluminum oxide, is used. Spots of solution are applied onto a base line near the slide’s lower edge, and the slide is then placed into a shallow bath of solvent. The solvent travels upward by capillary action, passing through the point at which the sample was applied, carrying the component with it at different rates. The T.L.C. plate is developed with aniline pthylate spray and viewed under ultraviolet light to reveal each component separated by T.L.C cards (Fig. 1). After the development of the silica gel card, the card was divided into six portions with respect to the spots, each spot was separated from other by cutting the silica card in form of silica strips, then each strip was cut into small pieces which were then dip into diethyl ether to extract the individual spot into the solvent and solvent was air dried and finally all the silica gel got dissolved. Now filter paper strips were dipped in the purified extracts along with the positive control i.e. the pure culture and negative control i.e. the silica gel dissolved in the diethyl ether. These filter paper strips were placed on the plate having bacterial lawn of Staphylococcus aureus and it was observed that within six hours the zones of inhibition were produced. And it was observed that two spots showed antibacterial activity not all. T.L.C of crude extract from CMG646 showed six spots with different Rf values (Table 4) three of which were aniline pthylate spray sensitive and three were U.V sensitive (Fig. 1) and two of which showed antibacterial activity upon inoculation in the plate having staphylococcus aureus lawn. As the bacterial strains CMG646, CMG645, CMG649 showed antibacterial activity against different indicator strains with the same intensity, so it seems that they show broad-spectrum antagonistic antibacterial activity and hence they are the producers of strong antibacterial compound. Fig. 3 Bioabsorbent Production As a Function of Time 3.5 3 2.5 Weight (grams) 2 1.5 1 0.5 0 0 50 100 150 200 250 Time (hours) Wet Gelatin Dry Gelatin Water Regained Copyright © 2011 SAVAP International www.savap.org.pk www.journals.savap.org.pk 131
Academic Research International ISSN: 2223-9553 Volume 1, Issue 2, September 2011 LITERATURE CITED 1. Agriculture statistics of Pakistan, 2001-2002. 2. Aslop, R. M. 1983. Industrial production of dextrans. Elsevier Science publishers. Vol. 83 printed in Netherlands. 40-44 3. Bonsall, R. F., Weller, D.M. and Thomashow S. L. 1997. Quantification of 2,4- Diacetylphloroglucinol Produced by Fluorescent Pseudomonas of Wheat. Appl. Eviron. Microbiol.63 (3): 951-955. 4. Burd, G. L., Dixon, D.G. and Glick, B.R. 2000. Plant growth promoting Rhizobacteria that decreases heavy toxicity in plants. Can. J. Microbiol. 33:237-245. 5. Carpitia, N., and C. Vergara 1998. Arecipe of cellulose. Science vol 279, 30-32. 6. Colwell, R.R. (1997). Microbial biodiversity and biotechnology. In: Biodiversity II: Understanding and protecting our biological resources. Reaka-Kudla, M.L., Wilson, D.E. & Wilson, E.O. (eds). Joseph Henry Press, University of Washington, D.C. 7. De Weger, L.A., Vaneder, A. J., Dekkers, L.C., Simons, M., Wijffelman, C.A., and Lugtenberg, B., 1995. Colonization of the rhizosphere of the crop plants by plant beneficial Pseudomonas. FEMS Microbiol. Ecol. 17:221-228. 8. Fasim, F., N. Ahmed and G. M. Gadd. 2002. Solubilization of zinc oxide and zinc phosphate by bacterium from air environment of a tannery. FEMS Microbiology Letters, 213 (1): 1-6. 9. Gale N.L.1981. The Role of Algea and other Microorganisms in Metal Detoxification. In Biotechnology for Mining. Metal refining and Fossil Fuel Processing Industries. Pp; 171-180. Jhon Wiely and sons, New York. 10. Hill, G. T., Motrowski, N.A., Wolfe, LA, Emele, L.R., Jurkoine, D.D., Ficke, A., Ramirez, S.M., Lynch, S.T. and Nelson, E.B., 2000. Methods for assessing the composition and diversity of soil microbial communities. Appl. Soil. Ecol., 15: 25-36. 11. http://www.nrdc.org. 12. http://www.agr.kuleuven.ac.be Copyright © 2011 SAVAP International www.savap.org.pk www.journals.savap.org.pk 132
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Vol. 1(2) SEP 2011 - SAVAP International

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