Difference between revisions of "Part:BBa K5443003:Experience"
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===Applications of BBa_K5443003===
 
===Applications of BBa_K5443003===
  
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We showed that the HpaBC enzyme was functional in E.coli as part of a vanillin biosynthetic pathway, as evidenced by production of caffeate (see LC-MS data from one clone pMQ3C-11). Further evidence for functional expression of the HpaBC monooxygenase was seen from production of a brown melanin-type dye as a side-reaction; we believe this indicates that HpaBC is acting on tyrosine (to make L-DOPA, which polymerises) in addition to our intended reaction on 4-coumarate to make caffeate.  
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We showed that the HpaBC enzyme was functional in <i>E. coli</i> as part of a vanillin biosynthetic pathway, as evidenced by production of caffeate (see LC-MS data from one clone pMQ3C-11, Fig. 1). Further evidence for functional expression of the HpaBC monooxygenase was seen from production of a brown melanin-type dye as a side-reaction (see Fig. 2); we believe this indicates that HpaBC is acting on tyrosine (to make L-DOPA, which polymerises) in addition to our intended reaction on 4-coumarate to make caffeate.  
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<figure>
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    <img src="https://static.igem.wiki/teams/5443/pmq3c11-lcms-data-v2.jpg" style="width:600px;height:auto">
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    <figcaption>
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        <b>Figure 1. LCMS analysis of compounds produced using pMQ3C-11.</b><br>
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        <i>The graph shows the detected presence of all 6 tested compounds, validating successful expression of all parts within pMQ3C-11.</i>
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    </figcaption>
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</figure>
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</html>
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<html>
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<figure>
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    <img src="https://static.igem.wiki/teams/5443/melanin-pmq3c.jpg" style="width:600px;height:auto">
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    <figcaption>
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        <b>Figure 2. Patched clones of plasmid variants.</b><br>
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        <i>Each plate shows patched colonies transformed with variants of our created plasmids pMQ3B and pMQ3C. Some patched colonies show diffusion of a brown pigment into the surrounding agar, suggesting a side-reaction of the enzyme HpaBC that produces the brown pigment L-DOPA.</i>
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    </figcaption>
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</figure>
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</html>
  
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===User Reviews===
 
===User Reviews===
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Revision as of 09:57, 2 October 2024

Applications of BBa_K5443003

We showed that the HpaBC enzyme was functional in E. coli as part of a vanillin biosynthetic pathway, as evidenced by production of caffeate (see LC-MS data from one clone pMQ3C-11, Fig. 1). Further evidence for functional expression of the HpaBC monooxygenase was seen from production of a brown melanin-type dye as a side-reaction (see Fig. 2); we believe this indicates that HpaBC is acting on tyrosine (to make L-DOPA, which polymerises) in addition to our intended reaction on 4-coumarate to make caffeate.


Figure 1. LCMS analysis of compounds produced using pMQ3C-11.
The graph shows the detected presence of all 6 tested compounds, validating successful expression of all parts within pMQ3C-11.



Figure 2. Patched clones of plasmid variants.
Each plate shows patched colonies transformed with variants of our created plasmids pMQ3B and pMQ3C. Some patched colonies show diffusion of a brown pigment into the surrounding agar, suggesting a side-reaction of the enzyme HpaBC that produces the brown pigment L-DOPA.


User Reviews

UNIQe114ea317e7c619a-partinfo-00000002-QINU UNIQe114ea317e7c619a-partinfo-00000003-QINU