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Team:Bielefeld-Germany/Project/Achievments
From 2012.igem.org
Project Achievements
- We successfully produced purified and identified active laccases of E. coli BL21(CueO named ECOL), Bacillus pumilus DSM 27 laccase (CotA named BPUL), Bacillus halodurans C-125 laccase (Lbh1 named BHAL) and Thermus thermophilus HB27 (tthL named TTHL).
- We successfully designed a shuttle vector for site-directed integration in P. pastoris which allows the production and secretion of a protein of interest. We were able to show that the vector is functioning as expected.
- We successfully produced the laccase TVEL5 from Trametes versicolor in P. pastoris and showed the activity of the produced enzymes.
- Results
- Successful characterization of the activity of all four bacterial laccases regarding the oxidization of ABTS under different conditions.
- We established a method for the degradation of several compounds with the help of our self-produced laccases. The HPLC analysis showed that our produced bacterial laccases are able to degrade estradiol with and without addition of ABTS. BPUL is able to degrade ethinyl estradiol with addition of ABTS. LC-MS measurements showed degradation products which could not be identified but were further analyzed using MS/MS. The resulting products have been discussed with an organic chemist.
- We successfully immobilized Trametes versicolor laccase (named TVEL0), E. coli BL21 laccase (CueO named ECOL) and Bacillus pumilus DSM 27 laccase (CotA named BPUL) and measured their activity.
- We successfully improved a BioBrick by clarifying its true function.
- [http://partsregistry.org/Part:BBa_K392014:Experience#User_Reviews Results]
- We successfully created our own database to organize our samples.
- We successfully created a model which simulates the oxidation of ABTS.
- We conducted a feasibility study of our project with sewage plant experts.
- We did an outstanding outreach work.
- We won a gold medal.
- We qualified for the Boston finals
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