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

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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


Problem

  • 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

  • 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

http://www.biw.kuleuven.be/dtp/cmpg/pgprb_images/PseudoGreen.jpg


Hypothesis

  • 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


Objectives

  • 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.


Variables


Apparatus and Materials


Procedures

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

Phase 1: Culturing the bacteria


Procedures

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

  • 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.


Procedures


Procedures

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


Procedures

Part III: Determining optimum pH for chelation


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

  • 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.


Timeline


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.


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


Thank you.


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