title: Experimental and computational optimization of an Escherichia coli co-culture for the efficient production of flavonoids.
abstract: Metabolic engineering and synthetic biology have enabled the use of microbial production platforms for the renewable production of many high-value natural products. Titers and yields, however, are often too low to result in commercially viable processes. Microbial co-cultures have the ability to distribute metabolic burden and allow for modular specific optimization in a way that is not possible through traditional monoculture fermentation methods. Here, we present an Escherichia coli co-culture for the efficient production of flavonoids in vivo, resulting in a 970-fold improvement in titer of flavan-3-ols over previously published monoculture production. To accomplish this improvement in titer, factors such as strain compatibility, carbon source, temperature, induction point, and inoculation ratio were initially optimized. The development of an empirical scaled-Gaussian model based on the initial optimization data was then implemented to predict the optimum point for the system. Experimental verification of the model predictions resulted in a 65% improvement in titer, to 40.7±0.1mg/L flavan-3-ols, over the previous optimum. Overall, this study demonstrates the first application of the co-culture production of flavonoids, the most in-depth co-culture optimization to date, and the first application of empirical systems modeling for improvement of titers from a co-culture system.
authors: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: jonesj12@rpi.edu.
authors: J Andrew
authors: JA
authors: Jones
authors: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: vernav@rpi.edu.
authors: Victoria R
authors: VR
authors: Vernacchio
authors: Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: sinkoa@rpi.edu.
authors: Andrew L
authors: AL
authors: Sinkoe
authors: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: collis6@rpi.edu.
authors: Shannon M
authors: SM
authors: Collins
authors: Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Chemistry of Natural Products Department, National Research Centre, Al-Bohoos St., 12622 Cairo, Egypt. Electronic address: ibrahm3@rpi.edu.
authors: Mohammad H A
authors: MHA
authors: Ibrahim
authors: Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: lachad2@rpi.edu.
authors: Daniel M
authors: DM
authors: Lachance
authors: Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: hahnj@rpi.edu.
authors: Juergen
authors: J
authors: Hahn
authors: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. Electronic address: koffam@rpi.edu.
authors: Mattheos A G
authors: MAG
authors: Koffas
mesh_terms: D018920:Coculture Techniques / Q000379:methods*; D003198:Computer Simulation; D004926:Escherichia coli / Q000254:growth & development*; D005419:Flavonoids / Q000096:biosynthesis*; D008954:Models, Biological
chemical_list: D005419:Flavonoids; C404987:flavan-3-ol
keywords: Co-culture; Flavan-3-ols; Microbial consortium; Pathway optimization; Systems modeling