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Team:Bielefeld-Germany/Project/Achievments
From 2012.igem.org
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| - | ::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 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). |
::[https://2012.igem.org/Team:Bielefeld-Germany/Results/coli Results ECOL] | ::[https://2012.igem.org/Team:Bielefeld-Germany/Results/coli Results ECOL] | ||
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| - | :: | + | ::We successfully characterized the activity of all four bacterial laccases produced regarding the oxidization of ABTS under different conditions. |
::[https://2012.igem.org/Team:Bielefeld-Germany/Results/coli#Activity_Analysis_of_ECOL Results ECOL] | ::[https://2012.igem.org/Team:Bielefeld-Germany/Results/coli#Activity_Analysis_of_ECOL Results ECOL] | ||
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::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 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. | ||
| - | ::[https://2012.igem.org/Team:Bielefeld-Germany/Results/ | + | ::[https://2012.igem.org/Team:Bielefeld-Germany/Results/substrate Results] |
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::[https://2012.igem.org/Team:Bielefeld-Germany/Results/immo Results TVEL0] | ::[https://2012.igem.org/Team:Bielefeld-Germany/Results/immo Results TVEL0] | ||
| - | ::[https://2012.igem.org/Team:Bielefeld-Germany/Results/immo | + | ::[https://2012.igem.org/Team:Bielefeld-Germany/Results/immo#Immobilization_of_purified_ECOL.2C_BPUL.2C_BHAL_and_TTHL_and_Activity_Tests Results ECOL, BPUL, BHAL and TTHL] |
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[[File:Bielefeld2012_haken_grau.jpg|30px|frameless|left]] | [[File:Bielefeld2012_haken_grau.jpg|30px|frameless|left]] | ||
| - | ::We successfully created a model which simulates the required laccase amount to | + | ::We successfully created a model which simulates the required laccase amount to degrade harmful substances under real sewage plant conditions. |
::[https://2012.igem.org/Team:Bielefeld-Germany/Modell Results] | ::[https://2012.igem.org/Team:Bielefeld-Germany/Modell Results] | ||
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| - | :::We won a gold medal. | + | :::We won a gold medal at the Europe Regional Jamboree. |
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[[File:Bielefeld-Germany2012_Qualified_stemp.png|60px|frameless|left]] | [[File:Bielefeld-Germany2012_Qualified_stemp.png|60px|frameless|left]] | ||
| - | :::We qualified for the world | + | :::We qualified for the world campionship |
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Latest revision as of 23:33, 2 December 2012
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
- We successfully characterized the activity of all four bacterial laccases produced 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 created a model which simulates the required laccase amount to degrade harmful substances under real sewage plant conditions.
- We conducted a feasibility study of our project with sewage plant experts.
- We successfully created our own database to organize our samples.
- We successfully improved a BioBrick by clarifying its true function.
- [http://partsregistry.org/Part:BBa_K392014:Experience#User_Reviews Results]
- We did an outstanding outreach work.
- We won a gold medal at the Europe Regional Jamboree.
- We qualified for the world campionship
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