Purification of Enzymes
We have successfully purified the enzymes needed. Below is the SDS-PAGE result of most of these enzymes.
From 1 to 10: Glk, Pgi, PfkA, FbaA, TpiA, Pgk, GpmA, Eno, PykF, GapA (M: protein ladder)
Measurement of Enzyme Concentration
We used Pierce™ Microplate BCA Protein Assay Kit to measure concentrations of each enzyme.
In Vitro Expression
We used protein expression kit by ThermoFisher Scientific © to express part Bba_J04450 along with the purified enzymes. Unfortunately, we failed to acquire a convincing result. This is possibly due to the inappropriate ratio of different components. We will continue to test different settings to test our designed glucose sensor.