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Latest revision as of 20:19, 19 October 2016
TEST what you can't TASTE.
Project
1.0 We engineer our bacteria to generate Penicillin Binding Protein 5-Green fluorescence Protein (PBP5-GFP), and bind penicillin to the coated wells on the ELISA plate. The protein is added to the sample, where the penicillin in the sample compete for PBP5-GFP with those on the plate, and the fluorescence intensity in the sample is measured and reflects its penicillin concentration.
2.0 PBP5 causes the hydrolysis of tripeptide Lys-Ala-Ala(KAA) when active and not bound with penicillin, releasing one alanine and exposing one amino group.[3] We measure its hydrolysis by the amino group, which changes the color of KAISER, a substance that reacts with free amino acids to yield CO2, NH3, and an aldehyde, the NH3 further yielding a colored product (diketohydrindylidene-diketohydrindamine, a bi-indanedione derivative also called Ruhemann purple), yielding the color purple. With more penicillin present, more PBP5 becomes inactive, and the purple gets lighter.