Journal:

Article Title: Improvement of Posttranslational Bottlenecks in the Production of Penicillin Amidase in Recombinant Escherichia coli Strains

doi: 10.1128/AEM.69.2.1237-1245.2003

Figure Lengend Snippet: E. coli strains used in this study

Article Snippet: Pure PA at a concentration of 0.1 mg/ml (B chain) and the slowly processed Thr263Gly mutant PA precursor (pPA) at a concentration of 0.06 mg/ml were used as standards. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Plasmid E. coli host Medium PA expression level (BPU/g of cell dry wt) None ATCC 11105 LB 80 M9 140 ρPAEC DH5α LB 50 M9Y 130 K5 LB 130 M9Y 295 M9 364 MC4100 LB 147 M9Y 250 M9 390 JM109 LB 190 M9Y 455 BL21(DE3) LB 110 M9Y 610 M9 1,170 b Open in a separate window a PA activity was measured 6 h after induction with 0.5 mM IPTG.

Techniques:

Journal:

Article Title: Improvement of Posttranslational Bottlenecks in the Production of Penicillin Amidase in Recombinant Escherichia coli Strains

doi: 10.1128/AEM.69.2.1237-1245.2003

Figure Lengend Snippet: PA expression levels in various E. coli host cells a

Article Snippet: Pure PA at a concentration of 0.1 mg/ml (B chain) and the slowly processed Thr263Gly mutant PA precursor (pPA) at a concentration of 0.06 mg/ml were used as standards. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Plasmid E. coli host Medium PA expression level (BPU/g of cell dry wt) None ATCC 11105 LB 80 M9 140 ρPAEC DH5α LB 50 M9Y 130 K5 LB 130 M9Y 295 M9 364 MC4100 LB 147 M9Y 250 M9 390 JM109 LB 190 M9Y 455 BL21(DE3) LB 110 M9Y 610 M9 1,170 b Open in a separate window a PA activity was measured 6 h after induction with 0.5 mM IPTG.

Techniques: Expressing

Journal:

Article Title: Improvement of Posttranslational Bottlenecks in the Production of Penicillin Amidase in Recombinant Escherichia coli Strains

doi: 10.1128/AEM.69.2.1237-1245.2003

Figure Lengend Snippet: In vivo proteolysis of ppPA in E. coli host cells. E. coli BL21(DE3) (A), E. coli MC4100 (B), and E. coli K5 (C) were used as host cells. Residual concentrations of ppPA and the 50-kDa proteolytic product were determined by immunoblot analysis after the inhibition of de novo protein synthesis by rifampin. The relative optical densities (dens.) of the bands of interest from the Western blot were quantified by using ONE-Dscan. ppPA density was related to the density of the pPA standard, and the density of the 50-kDa proteolytic band was related to the density of the PA standard (for details, see Materials and Methods). LB, LB medium; M9, M9 minimal medium.

Article Snippet: Pure PA at a concentration of 0.1 mg/ml (B chain) and the slowly processed Thr263Gly mutant PA precursor (pPA) at a concentration of 0.06 mg/ml were used as standards. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Plasmid E. coli host Medium PA expression level (BPU/g of cell dry wt) None ATCC 11105 LB 80 M9 140 ρPAEC DH5α LB 50 M9Y 130 K5 LB 130 M9Y 295 M9 364 MC4100 LB 147 M9Y 250 M9 390 JM109 LB 190 M9Y 455 BL21(DE3) LB 110 M9Y 610 M9 1,170 b Open in a separate window a PA activity was measured 6 h after induction with 0.5 mM IPTG.

Techniques: In Vivo, Western Blot, Inhibition

Journal:

Article Title: Improvement of Posttranslational Bottlenecks in the Production of Penicillin Amidase in Recombinant Escherichia coli Strains

doi: 10.1128/AEM.69.2.1237-1245.2003

Figure Lengend Snippet: Rate constants for intracellular proteolysis and transport in different strains a

Article Snippet: Pure PA at a concentration of 0.1 mg/ml (B chain) and the slowly processed Thr263Gly mutant PA precursor (pPA) at a concentration of 0.06 mg/ml were used as standards. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Plasmid E. coli host Medium PA expression level (BPU/g of cell dry wt) None ATCC 11105 LB 80 M9 140 ρPAEC DH5α LB 50 M9Y 130 K5 LB 130 M9Y 295 M9 364 MC4100 LB 147 M9Y 250 M9 390 JM109 LB 190 M9Y 455 BL21(DE3) LB 110 M9Y 610 M9 1,170 b Open in a separate window a PA activity was measured 6 h after induction with 0.5 mM IPTG.

Techniques:

Journal:

Article Title: Improvement of Posttranslational Bottlenecks in the Production of Penicillin Amidase in Recombinant Escherichia coli Strains

doi: 10.1128/AEM.69.2.1237-1245.2003

Figure Lengend Snippet: OmpT-PA expression levels in various E. coli host cells a

Article Snippet: Pure PA at a concentration of 0.1 mg/ml (B chain) and the slowly processed Thr263Gly mutant PA precursor (pPA) at a concentration of 0.06 mg/ml were used as standards. table ft1 table-wrap mode="anchored" t5 TABLE 2. caption a7 Plasmid E. coli host Medium PA expression level (BPU/g of cell dry wt) None ATCC 11105 LB 80 M9 140 ρPAEC DH5α LB 50 M9Y 130 K5 LB 130 M9Y 295 M9 364 MC4100 LB 147 M9Y 250 M9 390 JM109 LB 190 M9Y 455 BL21(DE3) LB 110 M9Y 610 M9 1,170 b Open in a separate window a PA activity was measured 6 h after induction with 0.5 mM IPTG.

Techniques: Expressing