Investigation of Siderophores from Pseudomonas fluorescens

Members:Vamsi MekaJosh HasanShaun PngJohnny YeungInvestigation of the Use of Siderophores from Pseudomonas genus to chelate Heavy Metal ions6/11/2009ProblemArsenic poising in the water of the village of Bangladesh and formation of lung cancer in patients causes health problems (Renshaw et al. 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 environments2Background - siderophoresThe 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.jpg3HypothesisSiderophores will 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.

ObjectivesThe 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 genusTo find the temperature and pH which allows the greatest degree of chelation.

VariablesIndependentDependentControlledType of heavy metal ionFinal concentration of heavy metal ionsInitial ion concentrationTemperature of incubationInitial bacteria innoculumpHApparatus and MaterialsApparatusMaterialsSpectrophotometerPseudomonas fluorescens ATCC 948MicropipettesPseudomonas syringae Inoculating loop Copper (II) ionsOrbital shakerFerric ions CentrifugeNickel ions Lead(II) ions Petri dishesLB mediumMSG mediumProceduresPhase 1: Culturing the bacteriaPart I: Investigating which heavy metal ions can be chelated:

http://www.gbiosciences.com/EducationalUploads/EducationalProductsImages/mediumimages/LB%20Broth.jpgProceduresSet upMSG medium/broth (Is bacteria present?)Presence of heavy metal ion solution1MSG medium bacteria presentPresent2MSG broth bacteria absentPresent3MSG medium bacteria presentAbsentPhase 2: introducing siderophores to solution.Step 4: Prepare 3 solutions as follows:Set up 1: Experimental Set up; Set up 2 and 3: controlsSet up 2: Determine if concentration of heavy metal ions changes in absence of Pseudomonas bacteriaSet up 3: Determine if heavy metal ions affect growth of Pseudomonas bacteriaIn the experiment, only record the results for set ups 1 and 2 to compare the difference in heavy metal ion concentration.ProceduresFor set ups 1 and 2, ProceduresAn arbitrary standard curveProceduresPart II: Determining optimum temperature for chelation

ProceduresPart III: Determining optimum pH for chelationRepeat the same procedures for Part I for 5 sets for each type of bacteria, but during step 3, alter the pH to 5.2, 6.2, 7.2, 8.2, 9.2 for each set.At the end of step 10, plot a graph of difference in concentration/ppm vs pH to determine the optimum pH for chelation.Data collectionChange in concentration/ppmThe higher the change, the higher the rate of chelationStatistical testsDone before and after introduction of siderophores from Psudomonas bacteriap