Investigation of the use of siderophores from pseudomonas genus to chelate heavy metal ions
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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

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



Josh Hasan

Shaun Png

Johnny Yeung



  • Arsenic poisoning in the water of Bangladeshi villages and formation of lung cancer in patients caused health problems (Renshawet al. , 2002).

  • Heavy metal ions are not biodegradable nor thermodegradable and are toxic. (Tansupoet al. , 2009)

  • Metal pollution caused by sources such as motor vehicle emissions, sewage sludge applications, and manufacturing has led to formation of hazardous environments

Background siderophores
Background - siderophores

  • The property of siderophores in chelating ferric ions have long been known and utilized in scientific industries (Duckworth & Sposito, 2007)

  • Siderophores are produced by bacteria under iron-starved conditions


  • Siderophores from both Pseudomonas fluorescensand Pseudomonas syringaewill demonstrate chelation for nickel, copper(II), iron(III) and lead(II) ions

  • Temperature and pH for the greatest degree of chelation is 26 degrees Celcius and 7.2 respectively for both species of Pseudomonas


  • The objective of the experiment is

    • To investigate chelationof 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.


Part I: Investigating the heavy metal ions that can be chelated:

Phase 1: Culturing the bacteria


Phase 2: Preparation of beads

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.

Phase 2 preparation of sodium alginate beads
Phase 2: Preparation of sodium alginate beads

  • Mix set ups with sodium alginate solution

  • Put the mixture dropwise into calcium chloride solution to obtain beads with bacteria inside for set up 1 and 3 but not 2.


Part II: Determining optimum temperature for chelation - test only for metal ions that are chelated during Part I


Part III: Determining optimum pH for chelation

Data collection
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 concentration 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

Work distribution and safety issues
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.


  • 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.


  • 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