Team:DTU-Denmark/Description

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Project Inspiration and Description

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. That's where we enter the scene.

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This pie chart shows that 49% of all industrially produced enzymes are produced by filamentous fungi, 2% by yeast, 27% by miscellaneous Bacteria, and 22% by others.
This pie chart shows that of the promoters in the iGEM registry, 79% are from <i>E. coli</i>, 1.75% from <i>B. subtilis</i>, 4.50% from bacteriophages, 2.25% from miscellaneous procaryotes, 8% from yeast, and 4.5% from miscellaneous eukaryotes

Description

Filamentous fungi are responsible for 50% of all industrially produced enzymes and 10% of all known bioactive compounds, therefore this multi-billion dollar industry is a big part of everyday life. Yet, while this is impressive, filamentous fungi can be utilized in the production of many other compounds. [1][2]


Whilst filamentous fungi are hugely important in industry, a disproportionately small number of iGEM teams have worked with this group of organisms. The 2018 DTU iGEM team Hyphae Hackers exploited filamentous fungi in their project but realized that the accessibility of parts was lacking severely compared to that of bacteria and yeasts (e.g. Escherichia coli and Saccharomyces cerevisiae).

In the 15 years of iGEM, 2754 teams have participated but until last year only 5 had worked in filamentous fungi, and these accounted for less than 5% of the promoters in the iGEM registry.

The lack of filamentous fungal parts, therefore, resulted in great difficulties for the 2018 DTU iGEM team with transforming the fungi and achieving the desired expression. While several promoter libraries exist for bacteria and eukaryotic organisms such as yeast and mammalian cells, there are currently no such libraries for filamentous fungi in the scientific literature. [3]

The Technical University of Denmark harbors unique facilities and personnel with great expertise in working with synthetic biology in filamentous fungi. The department furthermore has a fermentation core which allows fungi to be examined in multiple scales and under differing conditions as well as control of the pH and monitoring of CO2-production during growth.


Promoters are essential in the production of metabolites, as they allow for the up or down-regulation of gene expression. The lack of fungal parts as well as the resources of the department inspired this year’s team to produce a library of synthetic promoters for filamentous fungi and other eukaryotic organisms. We built a piece of software that compared the genomes of several different species of Aspergillus to determine sequence conservation and create a library of synthetic promoters. The library provides promoters of different strengths that give consistent results between batches and scales.


Following discussions with several scientists and companies, the team decided on a strategy that would supply future iGEM teams with the tools required to do synthetic biology in filamentous fungi as well as close a gap in scientific literature, making our project useful in iGEM, academia, and industry.



(1) Novozymes, “The Novozymes Report 2017,” Novozymes A/S, 2016.
(2) S. Siddiqui, Protein Production: Quality Control and Secretion Stress Response. Elsevier B.V., 2016.
(3) http://parts.igem.org/Promoters/Catalog/Anderson, visited 16. october 2019

The logos of our three biggest supporters, DTU Blue Dot, Novo Nordisk fonden and Otto Mønsted fonden The logos of all of our sponsors, DTU, BioNordica, Eurofins Genomics, Qiagen, NEB New England biolabs, IDT Integrated DNA technologies and Twist bioscience