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Investigation of the Use of Siderophores from Pseudomonas genus to chelate Heavy Metal ions

Investigation of the Use of Siderophores from Pseudomonas genus to chelate Heavy Metal ions. Members: Vamsi Meka Josh Hasan Shaun Png Johnny Yeung. 6/11/2009. Problem.

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Investigation of the Use of Siderophores from Pseudomonas genus to chelate Heavy Metal ions

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  1. Investigation of the Use of Siderophores from Pseudomonas genus to chelate Heavy Metal ions Members: VamsiMeka Josh Hasan Shaun Png Johnny Yeung 6/11/2009

  2. Problem • Arsenic poising in the water of the village of Bangladesh and formation of lung cancer in patients causes health problems (Renshawetal. 2002). • Heavy metal ions are not biodegradable nor thermodegradable and are toxic. (Tansupo et al. , 2009) • Metal pollution caused by factors such as motor vehicle emissions, sewage sludge applications, and manufacturing has led to formation of hazardous environments

  3. Background - siderophores • The property of siderophores in chelating ferric ions have long been known and utilized in scientific industries (Duckworth & Sposito, 2007) • Contains antimicrobial properties (Barry & Challis, 2009). http://www.biw.kuleuven.be/dtp/cmpg/pgprb_images/PseudoGreen.jpg

  4. Hypothesis • Siderophores will demonstrate chelation for nickel, copper(II), iron(III) and lead(II) ions for both species of Pseudomonas • Temperature and pH for the greatest degree of chelation is 26 degrees Celcius and 7.2 respectively for both species of Pseudomonas

  5. Objectives • The objective of the experiment is • To investigate the degree of chelation of heavy metal ions of copper (II), iron (III), nickel and lead(II) by siderophores from Pseudomonas genus • To find the temperature and pH which allows the greatest degree of chelation.

  6. Variables

  7. Apparatus and Materials

  8. Procedures Part I: Investigating which heavy metal ions can be chelated: Phase 1: Culturing the bacteria

  9. Procedures Phase 2: introducing siderophores to solution. Step 4: Prepare 3 solutions as follows: Set up 1: Experimental Set up Set up 2: Determine if concentration of heavy metal ions changes in absence of Pseudomonas bacteria Set up 3: Determine if heavy metal ions affect growth of Pseudomonas bacteria In the experiment, only record the results for set ups 1 and 2 to compare the difference in heavy metal ion concentration.

  10. Procedures For set ups 1 and 2,

  11. Procedures Part II: Determining optimum temperature for chelation

  12. Procedures Part III: Determining optimum pH for chelation • Repeat the same procedures for Part I for 5 sets for each type of bacteria, but during preparation of MSG medium, alter the pH to 5.2, 6.2, 7.2, 8.2, 9.2 for each set. • After obtaining absorbance values, plot a standard graph to find the final concentration of the set up and thus calculate the change in concentration • Plot a graph of the change in concentration/ppmvs pH for various values of pH.

  13. Data collection • Change in concentration/ppm • The higher the change, the higher the rate of chelation • Graph plotting • Plot graphs for 3 different experiments • Change in concentration against type of metal • Change in concetration against temperature • Change in concentration against pH • Statistical tests • Conducted with concentration obtained before and after introduction of siderophores from Pseudomonas bacteria • p<0.05 suggests that the difference is significant and vice versa. • Sample size=5 • Paired t-test to be done

  14. Work distribution and safety issues • Both AOS and HCI will take exactly the same steps (same heavy metal ion, same treatment etc.) • However, AOS would work with Pseudomonas syringae while HCI to work with Pseudomonas fluorescens. Both are at BSL level 1 so they are safe to work with. • Lab coats and gloves are worn at all times during experiments.

  15. Timeline

  16. Progress • Collected Articles to use in Background Research • Background Research draft completed and peer reviewed by three people • Singaporean team has arrived in AOS and worked out details for project. • Both sides to work on their respective tasks while in AOS and HCI respectively and combine results and explanations when AOS arrives in HCI in August 2010. • A parallel project to ease combination in the future.

  17. References • Barry, S.M., & Challis G.L. (2009). Recent advances in siderophore biosynthesis. Current Opinions in Chemical Biology, 13(2), 205-215 • Cook, R. J. 1993. Making greater use of introduced microorganisms for biological control of plant pathogens. Annu. Rev. Phytopathol. 31:53–80. • Duckworth, O.W., & Sposito G. (2007). Siderophore-promoted dissolution of synthetic and biogenic layer-type Mn oxides. Chemical Geology, 242(3-4), 497-508 • Miethke, M., & Marahiel M.A. (2007). Siderophore-based iron acquisition and pathogen control. Microbiology and molecular biology reviews, 71(3), 413-451 • Nielands, J. B. (1995). Siderophores: structure and function of microbial iron transport compounds. The Journal of Biological Chemistry, 270(45), 26723-26726. • Renshaw, J.C., Robson, G.D., Trinci, A.P., Wiebe, M.G., & Livens, F.R. (2002). Fungal siderophores: structures, functions and applications. Mycological Research, 106(10), 1123-1142 • Visca, P., Imperi, F., & Lamont, I.L. (2006). Pyoverdinesiderophores: from biogenesis to biosignificance. TRENDS in Microbiology, 15(11), 22-30

  18. Thank you.

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