Difference between revisions of "Part:BBa K3829014"
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The composite Part is used to express MHETase. SS secretes MHETase out of the cell, while anchor protein 5105 anchor it on the cell surface. This part is based on<a href="https://parts.igem.org/Part:BBa_K3829011">BBa_K3829011</a>, replacing GFP with MHETase. | The composite Part is used to express MHETase. SS secretes MHETase out of the cell, while anchor protein 5105 anchor it on the cell surface. This part is based on<a href="https://parts.igem.org/Part:BBa_K3829011">BBa_K3829011</a>, replacing GFP with MHETase. | ||
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<P>In order to evaluate the surface display of MHETase, HPLC was used to detecte the enzyme activity. We set different reaction times, namely 10, 20, 30, 60, and 90 min. The HPLC results showed that the substrate decreased sharply within 60 min, and the degradation was almost complete at 60 min. Besides, MHET could not be detected at 90 minutes, indicating that the display of MHETase was successful.</P> | <P>In order to evaluate the surface display of MHETase, HPLC was used to detecte the enzyme activity. We set different reaction times, namely 10, 20, 30, 60, and 90 min. The HPLC results showed that the substrate decreased sharply within 60 min, and the degradation was almost complete at 60 min. Besides, MHET could not be detected at 90 minutes, indicating that the display of MHETase was successful.</P> | ||
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<br/><b>Fig.2</b> Determination of enzyme activity of MHETase. | <br/><b>Fig.2</b> Determination of enzyme activity of MHETase. | ||
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Revision as of 06:53, 21 October 2021
P-ss-MHETase-V5tag-Anchor protein 5105-T
The composite Part is used to express MHETase. SS secretes MHETase out of the cell, while anchor protein 5105 anchor it on the cell surface. This part is based onBBa_K3829011, replacing GFP with MHETase.
On the basis of BBa_K3829011, we replaced MHETase with yeGFP to obtain the composite parts BBa_K3829014. The structure was shown in figure 1.
Fig.1 The structure of the gene circuit.
In order to evaluate the surface display of MHETase, HPLC was used to detecte the enzyme activity. We set different reaction times, namely 10, 20, 30, 60, and 90 min. The HPLC results showed that the substrate decreased sharply within 60 min, and the degradation was almost complete at 60 min. Besides, MHET could not be detected at 90 minutes, indicating that the display of MHETase was successful.
Fig.2 Determination of enzyme activity of MHETase.
Sequence and Features