The 2018 Groningen IGEM team has produced several BioBricks that may be useful for future teams.
Phenylalanine ammonia lyase 2 (PAL2) from Arabidopsis thaliana catalyzes the deamination of phenylalanine. It was successfully used in our synthetic styrene production pathway in yeast. The enzyme was proved to function as expected by measuring trans-cinnamic acid production over time.
T7 PAL2 BBa_K2783011
A composite part with PAL2 was created, adding T7 expression functionality to the part. Successful expression was proven to by SDS-PAGE gel, which showed a strong band at the expected molecular weight.
Endoglucanase II from Trichoderma reesei cleaves the beta-1,4 glycosidic bond in cellulose, a resilient polymer of glucose. This part was used in our consolidated bioprocess to degrade cellulose into usable sugars. We showed that growth on cellulose was possible using several cellulase enzymes, and EGII itself was proven to be functional via a colorimetric assay.
T7 EGII BBa_K2783020
T7 expression functionality was added to our EGII part to allow easy expression in Escherichia coli. Functionality of this composite part was proven by western blotting using anti-his antibodies, which showed the presence of His-tagged protein after expression was induced.