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1 ampicillin  (Thermo Fisher)


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    Thermo Fisher 1 ampicillin
    In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.
    1 Ampicillin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 3912 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Engineering Escherichia coli for polyethylene terephthalate powder biodegradation via recoding of an outer membrane protein"

    Article Title: Engineering Escherichia coli for polyethylene terephthalate powder biodegradation via recoding of an outer membrane protein

    Journal: iScience

    doi: 10.1016/j.isci.2025.114621

    In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.
    Figure Legend Snippet: In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.

    Techniques Used: In Vivo, Cell Culture, Generated, Control, Concentration Assay, Expressing



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    In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.
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    In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.
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    In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.
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    In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.

    Journal: iScience

    Article Title: Engineering Escherichia coli for polyethylene terephthalate powder biodegradation via recoding of an outer membrane protein

    doi: 10.1016/j.isci.2025.114621

    Figure Lengend Snippet: In vivo accumulation of PET degradation products by genome-rewired E . coli cultured with PET powder (A) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of pPET (solid lines, primary axis) and the growth of strains S3 and control S1 during this experiment (dotted lines, secondary axis). Cultures (10 mL) were grown in 50 mL Falcon tubes for 192 h in M9 medium supplemented with 0.1 mg mL −1 ampicillin at 37°C and 200 rpm, with an initial OD 600 nm of 0.1 and a pPET concentration of 5.0 mg mL −1 . To induce growth via EG utilization, 1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and 0.1 mg mL −1 glycerol were added. We would like to note that in our previous work, strains expressing only the recoded PETase-like protein (LsrB m ) or only the EG-metabolizing genes did not grow, confirming their inability to use EG as a carbon source. These controls were therefore not repeated here; instead, strain S1, which carries both heterologous EG-metabolizing genes and the recoded periplasmic LsrB m , was used as the main control, as it can hydrolyze nanometric PET particles but not PET powder, as shown in this study. (B) Concentrations of degradation products generated during in vivo cultivation of strain S3 in the presence of different concentrations of pPET. Reactions were performed in 2-mL Eppendorf tubes using 500 μL of culture (OD 600 nm of 0.1) in M9 medium supplemented with 0.1 mg mL −1 ampicillin, 1 mM IPTG, 0.1 mg mL −1 glycerol, and different concentrations of pPET (5, 10, 20, and 30 mg mL −1 ) at 37°C and 1,200 rpm. (A and B) Absorbance in (A) was quantified spectrophotometrically, and the degradation products in (A and B) were quantified using HPLC, as detailed in . The values represent the means of three biological replicates ( n = 3), with the means and standard deviations (SDs) shown. Unprocessed data are available in D and S1F.

    Article Snippet: In brief, selected E . coli BL21 (DE3) clones were grown at 37°C on LB agar plates (ThermoFisher Scientific, MA, USA; ref. 22700041) supplemented with 0.1 mg mL -1 ampicillin (ThermoFisher Scientific; ref. BP1760).

    Techniques: In Vivo, Cell Culture, Generated, Control, Concentration Assay, Expressing