Besides designing a bioengineered microorganism that could help solve the methanol poisoning problem, the pragmatically goal on wet lab was to generate new data on PAOX1 and PFLD1 transcriptional profiles, measuring the performance of kinetic parameters and input compatibility using the standardization principles of synthetic biology [1]. We also tested some P. pastoris grow and cultivation variables to verify how it would behave on detection conditions and on the lyiophilization (freeze-drying) process.

The main questions regarding on our experimental approach are:

  • BioBrick Characterization Questions
    • How strong are the AOX1 and FLD1 promoters?
    • How relatively strong are the PAOX1 variants?
    • How different alcohol sources will behave on interaction with wild PAOX1 and variants? (Activation? Repression?)
    • Will PFLD1 be activated or repressed by inputs other than mentioned on literature (methylamine and methanol)?
    • How long is the time delay response of these promoters in a system with RFP?
    • Have our Pichia codon-optimized RFP a better fluorescence than a non-optimized (BBa_E1010)?

  • Microbiology Questions
    • Will P. pastoris survive on ethanol solutions?
    • Will P. pastoris survive lyophilization? If survives, will the survival population be large enough to deliver the output signal at naked eye!?

Our team managed to answer some of these questions and is still working to completely deliver all of them with good quality. You may start checking our results on our Assemblies page.

Back to our mathematical models Check out our Human Practices results