Difference between revisions of "Part:BBa K4949004"
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'''Figure 1.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, <i>E.coli</i> BL21 (DE3) + Est 119 or <i>E.coli</i> BL21 (DE3)ΔLpp + Est119, a native Lpp deficient stain, were washed and resuspended in PBS with 100µM NPO with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | '''Figure 1.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, <i>E.coli</i> BL21 (DE3) + Est 119 or <i>E.coli</i> BL21 (DE3)ΔLpp + Est119, a native Lpp deficient stain, were washed and resuspended in PBS with 100µM NPO with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | ||
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| − | <html><img src="https://static.igem.wiki/teams/4949/wiki/results/figure19-est119-pnob-deltalpp-strain.png" width="950" height=" | + | <html><img src="https://static.igem.wiki/teams/4949/wiki/results/figure19-est119-pnob-deltalpp-strain.png" width="950" height="450"></html> |
'''Figure 2.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, <i>E.coli</i> BL21 (DE3) + Est119 or <i>E.coli</i> BL21 (DE3)ΔLpp +Est119, a native Lpp deficient stain, were washed and resuspended in PBS with 100µM pNOB with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | '''Figure 2.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, <i>E.coli</i> BL21 (DE3) + Est119 or <i>E.coli</i> BL21 (DE3)ΔLpp +Est119, a native Lpp deficient stain, were washed and resuspended in PBS with 100µM pNOB with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | ||
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| − | <html><img src="https://static.igem.wiki/teams/4949/wiki/results/figure21-est119-npo-deltalpp-plasmid.png" width="950" height=" | + | <html><img src="https://static.igem.wiki/teams/4949/wiki/results/figure21-est119-npo-deltalpp-plasmid.png" width="950" height="450"></html> |
'''Figure 3.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression <i>E.coli</i> BL21 (DE3) + Est119 or <i>E.coli</i> BL21 (DE3) + Est119(ΔLpp), bacteria with a Lpp deficient plasmid, were washed, and resuspended in PBS with 100µM pNOB with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | '''Figure 3.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression <i>E.coli</i> BL21 (DE3) + Est119 or <i>E.coli</i> BL21 (DE3) + Est119(ΔLpp), bacteria with a Lpp deficient plasmid, were washed, and resuspended in PBS with 100µM pNOB with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | ||
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| − | <html><img src="https://static.igem.wiki/teams/4949/wiki/results/figure21-est119-npo-deltalpp-plasmid.png" width="950" height=" | + | <html><img src="https://static.igem.wiki/teams/4949/wiki/results/figure21-est119-npo-deltalpp-plasmid.png" width="950" height="450"></html> |
'''Figure 4.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, <i>E.coli</i> BL21 (DE3) + Est119 or <i>E.coli</i> BL21 (DE3) + Est119(ΔLpp), bacteria with an Lpp deficient plasmid, were washed and resuspended in PBS with 100µM NPO with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | '''Figure 4.''' Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, <i>E.coli</i> BL21 (DE3) + Est119 or <i>E.coli</i> BL21 (DE3) + Est119(ΔLpp), bacteria with an Lpp deficient plasmid, were washed and resuspended in PBS with 100µM NPO with a cell density normalized to OD<sub>600</sub>=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm. | ||
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Revision as of 03:04, 12 October 2023
Est119
Est119 is an esterase originally identified in the Thermobifida alba strain AHK119 (AB298783). Est119 has been shown to degrade aliphatic-aromatic copolyesters and decrease the size of polymer particles of other biodegradable plastics, with an optimal temperature range of 45-55°C (Hu and al. 2009). Similarly to MGS0156, Est119 is interesting due to its potential temperature compatibility with Manitoban composting methods. Our team modified the Est119 genetic sequence to include the Lpp-OmpA anchor to allow for the characterization of the surface-display mechanism of PLAnet Zero
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Figure 1. Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, E.coli BL21 (DE3) + Est 119 or E.coli BL21 (DE3)ΔLpp + Est119, a native Lpp deficient stain, were washed and resuspended in PBS with 100µM NPO with a cell density normalized to OD600=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm.
Figure 2. Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, E.coli BL21 (DE3) + Est119 or E.coli BL21 (DE3)ΔLpp +Est119, a native Lpp deficient stain, were washed and resuspended in PBS with 100µM pNOB with a cell density normalized to OD600=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm.
Figure 3. Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression E.coli BL21 (DE3) + Est119 or E.coli BL21 (DE3) + Est119(ΔLpp), bacteria with a Lpp deficient plasmid, were washed, and resuspended in PBS with 100µM pNOB with a cell density normalized to OD600=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm.
Figure 4. Absorbance as a function of time for the whole cell esterase activity assay of Est119. After induced expression, E.coli BL21 (DE3) + Est119 or E.coli BL21 (DE3) + Est119(ΔLpp), bacteria with an Lpp deficient plasmid, were washed and resuspended in PBS with 100µM NPO with a cell density normalized to OD600=1.0. Samples were then incubated for 5 min at room temperature. Aliquots of 100 µl were collected at 5 intervals and the absorbance of the supernatant as measured at 410 nm.