Difference between revisions of "Team:DTU-Denmark/Description"
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Revision as of 06:21, 20 October 2019
Project Inspiration and Description
The total annual revenue from enzymes produced by Aspergillus spp. is 480 mio USD per year [3].
Description
Filamentous fungi are used as cell factories for approximately 28% of all known bioreactive microbial metabolites [1]. With their ability to increase or decrease the expression of a metabolite, promoters are essential in the production of bioproducts.
Several promoter libraries exist for bacteria and eukaryotic organisms such as yeast and mammalian cells. Nevertheless, there are currently no such libraries for filamentous fungi in the scientific literature.
Our project aims to create a constitutive promoter library for filamentous fungi and other eukaryotic organisms that provide promoters of different strengths that give reproducible results between batches and scales.
The total annual revenue from enzymes produced by Aspergillus spp. is 480 mio USD per year [3].
Inspiration
The inspiration for the project came from several places that together created the goal we have now. First, the iGEM team from DTU in 2018 worked with filamentous fungi and it became clear that only very few parts in the registry have been used in filamentous fungi. This resulted in many difficulties, getting the system to work and getting anything expressed. This year we will fill the hole in the registry with parts that work in filamentous fungi and we will provide information on how to successfully work with filamentous fungi, including protoplastation, transformation, and product assays.
The second inspiration came from our supervisors’ colleagues in academia and contacts in the industry. Here promoter libraries are very useful, but the promoters in literature is limited and spread out. We saw a hole that we could fill and provide a valuable product to researchers across the world.
Lastly, at the Technical University of Denmark, we have unique facilities and personnel who have expertise in working with synthetic biology in filamentous fungi. The research being done in filamentous fungi, most specifically Aspergillus allows us to attempt this project as these organisms are harder to work with than bacteria. Furthermore, the department has a fermentation core that we can use to examine the strength of the promotes in multiple scales and in differing conditions.
Taking all of this together, we see that our project is a combination between introducing a new organism to iGEM with the tools required to do synthetic biology, filling a hole in the literature, and the unique opportunity we have.
Pick your Promoter
An12g07450
Alias: PmstA
mstA encodes a high-affinity sugar transporter, and has been shown to be expressed when A. niger starts to run out of sugar. It doesn't care about the age/phase of the culture, only about sugar
Special Ability: sensitive sugar sensor, turns on when sugar is limited
Versions: 1 naturalistic & 1 synthetically enhanced
An07g00070
A homolog of the A. fumigatus tpcK gene, part of the synthesis pathway for a mycotoxin, this gene represents a classical secondary metabolite, and its promoter acts accordingly: with relatively low leakiness, it turns on hard when the fungi reach stationary phase, a boon for growth-decoupled industrial production.
Despite the scary story of the gene it regulates, the promoter is perfectly tame, harmless, and ready to help.
Versions: 6 versions, with 4 levels: synthetically enhanced 1x, naturalistic-ish 2x, sorta weakened 2x, even more weakened 1x
(1) J. Berdy, “Bioreactive Microbial Metabolites” J. antibiot, vol. 58. no. 1, pp. 1-26, 2005.
