Fig. 6: Engineering dynamic pathway regulation for DHS production using GPTAC.
From: De novo designed protein guiding targeted protein degradation
a Schematic representation of GPTAC-controlled metabolic flux regulation. In the DHS production phase, GPTAC expression not only continuously degraded AroE expressed from the genome but also rapidly eliminated previously expressed AroE, enabling a swift blockage of metabolic flux and facilitating DHS accumulation. b Binding interface selection (Left) and design models (Right) for GP complexes. Left: AroE surface showing hydrophobic residues (red) and Patchdock-selected binding sites (yellow). Right: Designed complexes of GPAroE-AroE, with binding sites (yellow) and targets (gray). c A dose-escalation experiment demonstrated GPTAC-mediated AroE degradation after one hour of treatment with different concentrations of IPTG (0.05 mM, 0.1 mM, 0.2 mM). d Detection of AroE degradation during the growth of E. coli DHS01 in M9 medium without essential amino acids. Data are reported as the average of three independent repeats. e Effect of continued addition of aTc or IPTG at 40 h on DHS production by E. coli DHS01. The first addition of the inducer occurred at 24 h. Furthermore, a decrease in DHS accumulation rate was observed at 40 h, prompting a second addition of the inducer at that time. All experiments are presented as mean ± s.d. from three biologically independent replicates. Source data are provided as a Source Data file. (a) is created in BioRender https://BioRender.com/a1rvfbu.
