ISBN 9783843946650

978-3-8439-4665-0, Reihe Biotechnologie

Minliang Chen
Novel approaches for in vivo evolution, screening and characterization of enzymes for metabolic engineering of Escherichia coli as hyper L-tryptophan producer

157 Seiten, Dissertation Technische Universität Hamburg (2020), Softcover, A5

Zusammenfassung / Abstract

Nowadays, microbes have been extensively optimized for the production of L-tryptophan (Trp) from renewable feedstocks. The main objective of this work was therefore to design and construct Trp producing E. coli strain(s) with more robust growth and more efficiency in Trp production and yield by the engineering of key enzymes (e.g., TrpC, AroG, and GalP-Glk). To effectively in vivo screen and characterize the enzyme variants, three approaches of linking growth-coupled and sensor-guided in vivo screening (PGSS, CGSS, and Auto-CGSS) were developed and examined for engineering enzyme variants with desired performances. Subsequently, integrative metabolic engineering was performed to assemble the enzyme variants into a Trp-producing strain S028.