Constitutive naringenin producing operon
Sascha Hein, Niklas Hummel, Sebastian Barthel, Thomas Dohmen
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).
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.
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
tyrosine ammonia-lyase (TAL) is an enzyme which catalyzes the formation of p-coumaric acid (aka p-hyroxycinnamic acid) from tyrosine. It belongs to the family of ammonia-lyases, enzymes that catalyze the deamination of amino acids [1]. P-coumaric acid is an important precursor in many metabolic pathways. TAL also has a secondary function as a phenylalanine ammonia-lyase (PAL) which catalyses the formation of cinnamic acid. The wild type RsTAL has a preference for tyrosine, however the mutations Met4 -> Ile, Ile325 -> Val and Val409 -> Met, can shift this preference to phenylalanine [2]. 1.Zhixiong Xue, Michael McCluskey, Keith Cantera, F. Sima Sariaslani, Lixuan Huang (2007) Identification, characterization and functional expression of a tyrosine ammonia-lyase and its mutants from the photosynthetic bacterium Rhodobacter sphaeroides. J Ind Microbiol Biotechnol 34:599-604 2. J.A. Kyndt, T.E. Meyer, M.A Cusanovich, J.J. Van Beeumen (2002) Characterization of a bacterial tyrosine ammonia lyase, a biosynthetic enzyme for the photoactive yellow protein. FEBS Letters 512 240-244
Earlier in iGEM eucaryot TAL has been used, however we used bacterial TAL for better compatibility with E.coli.
Karl Holdar, Hampus Elofsson, Emil Marklund, Kristoffer Lundmark, Marcus Hong, Lovisa Pettersson, Ken Braech-Andersen, Theodor L??we
4-coumarate--coenzyme A ligase (4CL) coding region
Plasmid (template for PCR) provided by John A. Morgan, School of Chemical Engineering, Purdue University, West Lafayette, Indiana, USA http://www.ncbi.nlm.nih.gov/nucleotide/609339/ PMID: 15932991
4-coumarate--coenzyme A ligase (4CL) coding region from ''Arabidopsis thaliana''.
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).
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).
The Chalcone Synthase catalyzes the reaction from 4-Coumaroyl-CoA to Naringenin Chalcone. It is one of the essential enzymes in the anthocyanin pathway.
Conversion of the iGEM parts registry to SBOL2.1