iGEM Marburg 2018: establishing Vibrio natriegens as a chassis for synthetic biology

Imagine a lab where you need twelve hours for a cloning cycle instead of three days. Imagine a lab where you start your culture in the morning and harvest your cells after lunch. Imagine a lab where the time lost waiting for cells to grow is reduced to the absolute minimum.

In the past years almost all processes (e.g. DNA synthesis or sequencing) have become faster, but one aspect is still unchanged: The organism that is used as chassis in the majority of Synbio projects remains Escherichia coli. The iGEM Team Marburg 2018 attempts to shake the fundaments of biological engineering by replacing E. coli with Vibrio natriegens, the fastest growing organism known to date, with a demonstrated doubling time of seven minutes.

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The iGEM Team Marburg and their project “Vibrigens - Accelerating Synbio“ aim for the establishment of V. natriegens as the new go-to organism for everyday lab work in both academic and industrial applications. This could improve production output of valuable chemicals and pharmaceuticals and pave the way for a brighter future of research. V. natriegens is very easy for labs to adapt to. It is a non-pathogenic S1 organism, which grows on salt rich standard LB-Medium.

The team is organized in three subgroups: First the strain engineering subgroup will adapt the genome to create the perfect chassis for molecular biology, as has been done with E. coli over the past decades. Strains without nucleases for ideal cloning, others with T7 expression systems and no proteases to express proteins as desired, and many more. These changes shall be done through genome engineering, recombineering and V. natrigens’s exceptional natural competence.

The part-collection subgroup will build the “Vibrigens MoClo toolbox” which will consist of a large number of precisely characterized parts. First experiments showed already, most parts commonly used in E. coli are also functional in V. natriegens. Still, plenty of work is required for the exact characterization of these parts in V. natriegens to achieve the same degree of predictability which is needed for the construction of synthetic regulatory networks or metabolic pathways.

Finally, the Metabolic Engineering subgroup will implement the first heterologous pathway into Vibrio natriegens, producing 3-Hydroxypropionic acid (3-HPA) one of the twelve most important bio-based chemicals needed to reduce dependence on fossil fuel. There is also another, theoretical pathway, which is the most efficient 3HPA-pathway. Because there is no known enzyme for the last reaction, it has never been tested in vivo, but that can be changed. The team will try to build multiple suitable enzymes by calculating the binding pocket for the substrate and subsequently expanding the protein structure until the whole enzyme is built. After testing these enzymes, the pathway can be perfected at a fast pace because biosensors will be established to translate product concentration into a measurable fluorescent output.

In their project “Vibrigens - Accelerating Synbio“ , Marburg's iGEM team is going to combine the fastest cloning methods available with the fastest growing organism Vibrio natriegens optimized for the needs of the community and to show its usefulness by producing 3-Hydroxypropionic acid.

In addition to their ambitious scientific project, iGEM Marburg will be hosting the German iGEM Meetup (22nd - 24th) in collaboration with iGEM Bielefeld and the GASB. This meetup will give all german iGEM teams a platform to reach out for collaborations and to present preliminary results of their projects.

 

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