Toxicity of Intermediates

Determine whether the ions nitrite, nitrate and ammonium are toxic to E. coli, and at what concentration. This experiment will use the Biolector to grow an E. coli strain that constitutively expresses RFP in the presence of these ions (at a range of concentrations) to determine the OD and fluorescence of the cells as a proxy for growth.

Toxicity Experiment

Verify nitrite stability in anaerobic, untransformed E. coli

Grow E. coli anaerobically in the presence of varying concentrations of nitrite; measure the amount of ammonia and nitrite present.

Experiment 1a

Measure production of N₂O from Nitrite NO₂^- anaerobically

Grow E. coli and P.aeroginosa anaerobically in the presence of varying concentrations of nitrite; measure the amount of nitrous oxide produced.

Experiment 2

Measure consumption of Ammonium in Mutant 1 transformants compared to native E. coli.

Grow AMO and HAO transformants aerobically in the presence of ammonia; measure concentration of ammonia.

Experiment 4

Growth of Nir transformant in Bioreactor

Bioreactor