Journal: Biotechnology Notes
Article Title: Model-based dynamic engineering of Escherichia coli for N-acetylglucosamine overproduction
doi: 10.1016/j.biotno.2022.02.001
Figure Lengend Snippet: Construction and application of the dynamic regulatory system. (A) The optimal flux of the reaction catalyzed by pfKAB and zwf in the simulated fermentation process. (B) Simulated optimal distribution of fluxes catalyzed by glmS and pfkAB , and specific growth rate during fermentation. (C) Schematic diagram of gene interference with CRISPRi system constitutive expression and results of shake flask fermentation. The dCas9 was under the control of P lacUV5 , and one sgRNA was under the control of P j23119 . (D) Schematic diagram and fluorescence verification results of the fluorescent reporter gene sf-GFP under the control of lacI and pdhR . The plasmid was transformed into E. coli JM109 , cultured in a 96-well plate with shaking at 37 °C for 8 h in LB liquid medium to detect the fluorescence intensity of the fermentation broth. Different concentrations of pyruvate and glucose were added with and without IPTG. (E) Schematic diagram and fermentation results of the dynamic regulation on pfkA and zwf under the control of pdhR and lacI . ** and * symbols indicate p < 0.01 and 0.01
Article Snippet: E. coli ATCC 25947 (DE3) , starting strain , Lab stock.
Techniques: Expressing, Control, Fluorescence, Plasmid Preparation, Transformation Assay, Cell Culture