Bastian Wagner, Christian Mende, Andreas Schmidt, Max Dombrowsky
B0034-CHI - Chalcone Isomerase from Petunia with strong RBS
Sascha Hein, Niklas Hummel, Sebastian Barthel, Thomas Dohmen
The Chalcone Isomerse catalyzes the reaction from Naringenin chalcone to Naringenin. This enzyme is non-essential for flavanone pathways, because the reaction takes place independently from CHI in the cytosol. The enzyme independent reaction is catalyzed by an acidic compound. Without CHI the reaction is stereo unspecific. S-Naringenin is the active compound for the most enzymes in the biochemical pathways. To prevent gaining a racemic product and losing 50% yield, CHI is used, because only the enzyme is able to form 100 % S-Naringenin. For using the CHI in our Naringenin Operon (K1497007) we added the strong RBS B0034.
B0034-CHS-B0034-CHI - Composite part of RBS-CHS and RBS-CHI
This device is able to generate Naringenin from 4-courmaroyl-CoA, if put under a promotor. This device is the second composite part for the assembly of our Naringenin Operon (K1497007). Part 1: K1497004: B0034-CHS - Chalcone Synthase from Gerbera with strong RBS The Chalcone Synthase catalyzes the reaction from 4-Coumaroyl-CoA to Naringenin Chalcone. It is one of the essential enzymes in the anthocyanin pathway. For using the CHI in our Naringenin Operon (K1497007) we added the strong RBS B0034. Part 2: K1497003: B0034-CHI - Chalcone Isomerase from Petunia with strong RBS The Chalcone Isomerse catalyzes the reaction from Naringenin chalcone to Naringenin. This enzyme is non-essential for flavanone pathways, because the reaction takes place independently from CHI in the cytosol. The enzyme independent reaction is catalyzed by an acidic compound. Without CHI the reaction is stereo unspecific. S-Naringenin is the active compound for the most enzymes in the biochemical pathways. To prevent gaining a racemic product and losing 50% yield, CHI is used, because only the enzyme is able to form 100 % S-Naringenin. For using the CHI in our Naringenin Operon (K1497007) we added the strong RBS B0034.
B0034-CHS - Chalcone Synthase from Gerbera with strong RBS
The Chalcone Synthase catalyzes the reaction from 4-Coumaroyl-CoA to Naringenin Chalcone. It is one of the essential enzymes in the anthocyanin pathway. For using the CHI in our Naringenin Operon (K1497007) we added the strong RBS B0034.
This composite part to produce naringenin from tyrosine, if put under a promotor. It consists of 4 genes: 4-CL (BBa_K801093) TAL (BBa_K1033000) CHI (BBa_K1497100) CHS (Bba_K1497101) For a strong and stable translation in front of each gene the RBS Bba_B0034 is implemented.
The functionality of this construct is verified by putting the device under a promotor. Proofing functionality leaded to Bba_K1497008 (Naringenin Operon under the constitutive promotor J23100) and Bba_K1497008 (Naringenin Operon under the T7 Promotor).
Pelargonidin producing operon B0034-F3H-B0034-DFR-B0034-eANS
Sascha Hein, Sebastian Barthel, Thomas Dohmen, Sven Jager
The Chalcone Isomerse catalyzes the reaction from Naringenin chalcone to Naringenin. This enzyme is non-essential for flavanone pathways, because the reaction takes place independently from CHI in the cytosol. The enzyme independent reaction is catalyzed by an acidic compound. Without CHI the reaction is stereo unspecific. S-Naringenin is the active compound for the most enzymes in the biochemical pathways. To prevent gaining a racemic product and losing 50% yield, CHI is used, because only the enzyme is able to form 100 % S-Naringenin. Reaction of the CHI. A proton shift of naringenin chalcone builds naringenin. We used the CHI and verified the function in our Naringenin Operon (K1497007).
Constitutive naringenin producing operon
The Chalcone Synthase catalyzes the reaction from 4-Coumaroyl-CoA to Naringenin Chalcone. It is one of the essential enzymes in the anthocyanin pathway.
Reaction of the CHS. 4-coumaroyl-CoA reacts together with 3 Malonyl-CoA to one Naringenin Calcone. We used the CHS and verified the function in our Naringenin Operon (K1497007).
Pelargonidin producing operon - T7-B0034-F3H-B0034-DFR-B0034-eANS
B0034-eANS (engineered anthocyanidin synthase with strong RBS)
Sascha Hein, Sven Jager, Thomas Dohmen, Sven Rumpf
Gene was E. coli optimated synthezied by Eurofins.
Naringenin producing operon under the T7 promoter BBa_I712074
Transformed into BL21(DE3) and induced with IPTG this composite is able to produce naringenin from tyrosine. It consists of 4 genes: 4-CL (BBa_K801093) TAL (BBa_K1033000) CHI (BBa_K1497000) CHS (BBa_K1497001) For a strong and stable translation in front of each gene the RBS Bba_B0034 is implemented. For characterization the naringenin biosensor K1497020 was used. After 24 h incubation GFP fluorescence was visible to the unaided eye. We measured the relative GFP fluorescence by use of OptimaFluoroStar.
Pelargonidin producing operon (low yield)- T7-B0034-F3H-B0034-DFR-B0034-ANS
Sascha Hein, Sebastian Barthel, Thomas Dohmen
This composite part is the fusion device of the Biobrick parts K1033001 by Uppsala 2013 and K1033000 by. It is able to produce 4-coumaroyl-CoA from tyrosine, if put under a promotor. This device is the first composite part for the assembly of our Naringenin Operon (K1497007). We used the previously announced Biobricks to assemble this part and verified the function in our Naringenin Operon (K1497007).