Approach

The conventional methods in sewage treatment plants are unable to treat waste water sufficiently because the most frequently used micro contaminants like estrogen, Bisphenol A, Dicolfenac etc. are very difficult to break down. The goal of Bielefeld’s iGEM team is to develop a biological filtersystem using immobilized laccases to purify municipal and industrial wastewater from synthetic estrogens and other aromatic compounds. Laccases are copper-containing oxidase enzymes found in many organisms, and one of their properties is the ability to break down a wide range of aromatic and phenolic compounds. For this purpose, genes of various bacterial and eukaryotic laccases should be isolated and expressed in Escherichia coli and Pichia pastoris. The choice of the expressionsystem depends on the glycosylation status of the enzyme.

The first step for Step of our project is to isolate the specific gene sequences and to generate new BioBricks for the iGEM competition. The Laccases of the following organisms are isolated: bactirial laccases:

1. Escherichia Coli
2. Baccilus Halodurans
3. Baccilus Pumilus
4. Streptomyces griseus
5. Streptomyces lavendulae
6. Themus thermophilus
7. Xanthomonas campestris

Eukaryotic laccases:

1. Arabidopsis thaliane
2. Tramestes Versicolor
3. Trametes villosa

For more information about the Organisms click here.

These BioBricks are used to design new plasmids and vector-systems. Plasmids, which are used for production in E.coli, includes a T7-Promotor, a ribosomal-binding-sequence ,a His-Tag-Sequence at the end of the Protein an a Chloramphenicol Resistent. The genetically modified organisms are used to produce different laccases.