TU Darmstadt Championship Poster
iGEM Team TU Darmstadt 2014 Linear Response Theory (LRT) TAL Introduction of a new ANS (eANS) with a shorter C-terminus leads to enhanced pelargonidin production F3´H CHI TAL 4CL ANS Residue position Problem region C4H F3H Residue position All models show a destabilized C-terminus Membrane DFR CHS Residue position Plants Heterologous systems CHI ΔRMSD The anthocyanidin pathway in plants is able to form a multi-enzyme complex. This complex increases the reaction efficiency of anthocyanidin synthase (ANS) [1]. In heterologous systems (e.g. E. coli) this complex is not formed and side reaction products are produced [2]. We identified and re-engineered the potential interaction site of the ANS and increased the yield of pelargonidin. Rigid core RMSF LRT Model of ANS Gaussian Network Molecular Dynamics Model (GNM) Simulation (MD) Slow modes Engineering and modeling of the anthocyanidin synthase E. grätzel F3H DFR CHS ANS 4CL Protein engineering Policy & Practices With engineered ANS more pelargonidin Naringenin Biosensor Naringenin Government / NGOs Purchasing machines Developing concepts Training Villagers Selling DSCs Cooperation Lending money Providing capital No access to electricity Modified E. coli produce anthocyanins DSC producer Grätzel cell construction Grätzel cells are an alternative for conventional Sensor solar cells. They generate construction electricity without rare earth metals. Application Small market for Grätzel cells Easy handling Adjustment of the pathway required! No glycosylation Extraction with organic solvent Grätzel cell Grätzel cell function chs R chi Dependent on the promotor the operon produces 1-3 µM naringenin. Naringenin is detectable and can be quantified. GFP fluorescense pH T7-BBa_K1497023 E. coli BL21(DE3) Fluorescence (RFU) R Plate with 100 µM Naringenin Control under UV light. mKate fluorescense pelargonidin is visible 4000 tal CFP fluorescense Anthocyanidin biosynthesis and production Control BBa_K1497007 R 3 Sensors with different fluorophores were constructed. Naringenin [µM] The implementation of the naringenin operon was necessary for pelargonidin production. 4-cl Solar cell Metabolic engineering Anthocyanidin precursor biosynthesis R gfp BBa_K1497019-22 Mutual learning Purchasing DSCs 30 kDa fdeR Dye Local partner Local microcredit bank FdeR Fluorescense [RFU] Local industry The goal: Harvesting solar power with anthocyanins Naringenin SDS-Page + - BBa_K1497023 3000 f3h R R dfr R eans 2000 Control T7–BBa_K1497007 1000 0 E. coli BL21(DE3) E. coli BL21(DE3) + BBa_K1497020 + BBa_K1497020 The biosynthesis of pelargonidin in E. coli is feasible. Pelargonidin can be extracted by dichloromethane. At pH 1 pelargonidin has a dark red color. Pelargonidin Naringenin Conclusions Full pelargonidin and naringenin biosynthesis pathways are deposited in the parts registry 3 new Biosensors for naringenin detection Successfull pelargonidin production and Grätzel cell construction 33 new Biobricks for the parts registry Novel mutual learning approach for Policy and Practices Refernces: [1] Yan Y, Chemler J, Huang L, [2] Winkel BSJ (2004) Metabolic channeling in et al. (2005) Metabolic plants. Engineering of Anthocyanin Biosynthesis in Escherichia coli. Our supporters: Special thanks to: A Prof. Dr. Michael Grätzel Prof. Dr. Katja Schmitz Dr. Toby Meier Dr. Stefan Martens Wieke Betten …and many more B
© Copyright 2025