Team:Bielefeld-Germany/Results/Datapage

From 2012.igem.org

Datapage


How our BioBricks work

Figure 1: The iGEM Team Bielefeld is developing a biological filter using immobilized laccases, enzymes able to oxidize and break down a broad range of aromatic substances. For the production of laccases from different bacteria, fungi and plants, three expression systems were used: Escherichia coli KRX and Rosetta-Gami 2 and the yeast Pichia pastoris. Immobilization is carried out either by using CPC-silica beads (Controlled Pore Carrier) or by fusing the enzymes to cellulose binding domains. The concept can be extended to other toxic pollutants in drinking and wastewater, as well as to industrial applications in paper and textile industries or even for bioremediation of contaminated soil.

Data for our favorite new parts

  1. BBa_K863000 - bpul (laccase from Bacillus pumilus) with T7 promoter, RBS and His-tag: This part is used to overexpress the laccase bpul for further purification followed by characterization of enzyme activity and substrate specificity.
  2. BBa_K863005 - ecol (laccase from E. coli) with T7 promoter, RBS and His-tag: This enzyme is overexpressed after induction with rhamnose and can be purified using its His-tag. Subsequently the laccase can be characterized regarding to its enzyme activity and substrate specificity.
  3. BBa_K863204 - shuttle vector pECPP11JS for site-directed recombination of genes of interest in yeast: With this part the production and secretion of a protein of interest like our laccase is possible. Protein expression is induced with methanol, selection is performed by complementation of its histidine auxotrophy.

We have also characterized the following parts

  1. BBa_K863012 - tthl laccase (from T. thermophilus) with constitutive promoter J23100, RBS and His-tag: This part is used for the constitutive expression of tthl for further purification followed by characterization of enzyme activity and substrate specificity.
  2. BBa_K863022 - bhal laccase (from Bacillus halodurans) with constitutive promoter J23100, RBS and His-tag: This part is used for the constitutive expression of bhal for further purification followed by characterization of enzyme activity and substrate specificity.
  3. BBa_K863207 - shuttle vector pECPP11JS for site-directed recombination of TVEL5 laccase in yeast: With this part the production and secretion of TVEL5 is possible.


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