Collaborations

Summary

IGEM is an international competition that promotes the search for solutions of problems in the world using synthetic biology. While every team is working on their own projects in their local laboratories utilizing the biobricks provided by iGEM over the summer, they design new parts to add to the collection of biobricks at the end of the competiton. This makes every team part of a big community, that works togehter to promote the transparent and innovative developement of new tools for engineering biology. This community is not only about the collection of biobricks, but also about helping each other. Collaborations of individual teams are one of the most important aspects of iGEM, where teams help each other achieve their goals. This can be just by giving helpful adivse or even doing experiments together. We collaborated intensively with the team of Marburg to determine the binding affinity of the EPSP synthases AroE and AroA to glyphosate.

Collaboration - Biochemical characterization of EPSP synthases in cooperation with iGEM Marburg

This year, our team came toghether with the iGEM team of Marburg, to investigate the interaction of the EPSP synthases AroE and AroA from B. subtilis and E. coli, respectively, with glyphosate. Glyphosate targets AroE and AroA in B. subtilis and E. coli, respectively. Since the members of the Bange lab, which is hosting the iGEM team of Marburg, are experts in the field of Isothermal titration calorimetry (ITC), we did not hesitate to contact them for support. Our team cloned the aroE and aroA genes from B. subtilis and E. coli, respectively, and the resulting constructs are suitable for overexpression of the N-terminally Strep-tagged AroE and AroA enzymes in E. coli. The proteins can be purified by Streptactin:Strep-tag affinity purification system from the IBA, Göttingen. The purified proteins were send to the iGEM team in Marburg (Figure 1). The ITC measurments were performed in Marburg. Unfortunately, not interaction between glyphosate and the EPSP synthases could be detected. Unfortunately, also the second ITC measurement using freshly purified AroA and AroE enzymes from E. coli and B. subtilis, respectively, did not reveal whether glyphosate may interact with the EPSP synthases. Nevertheless, we are grateful to the iGEM Team Marburg for performing the ITC measurements.

Figure 1. (A) Evaluation of the purification of the N-terminally Strep-tagged EPSP synthases from B. subtilis and E. coli Strep-AroE and Strep-AroA, respectively, by 12% SDS PAGE. The proteins were stained with Coomassie Brilliant Blue. M, unstained protein molecular weight marker Thermo Scientific; CE, crude extract; FT, flow through, W, washing steps; E, elution steps. (B) ITC measurements with Strep-AroE and Strep-AroA and glyphosate. (C) Control experiments with water and the calcium chelator EDTA.

For more information about the project of the iGEM Team Marburg 2018 please do not hesitate and visit their website: iGEM team of Marburg