Difference between revisions of "Part:BBa K5443013"
 
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Enoyl-CoA Hydratase/Aldolase (ECH) catalyzes the fifth and final step in the vanillin biosynthesis pathway, converting feruloyl-CoA to vanillin, thus completing the biosynthetic pathway. The enzyme belongs to the enoyl-CoA hydratase/isomerase superfamily and performs a two-step reaction, hydrating the double bond of feruloyl-CoA, then cleavage of the resulting thioester via a retro-aldol reaction. The ECH enzyme can also potentially use caffeoyl-CoA and 4-coumaroyl-CoA as substrates. We tested two FCS enzymes for this pathway step; this enzyme from Pseudomonas, and also a homolog from Streptomyces.
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The Enoyl-CoA Hydratase/Aldolase (ECH) enzyme from <i>Pseudomonas putida</i>, <b>ECH-Ppu</b>, catalyses the fifth and final step in our vanillin biosynthesis pathway. ECH-Ppu converts feruloyl-CoA to vanillin by performing a two-step reaction: it hydrates the double bond of feruloyl-CoA, then cleaves the resulting thioester via a retro-aldol reaction (Fig. 1). The ECH enzyme can also potentially use caffeoyl-CoA and 4-coumaroyl-CoA as substrates.
  
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This part builds on the work of earlier iGEM teams, who contributed ECH Parts as follows: Part:BBa K1129002
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We tested two alternate FCS enzymes for this pathway step; this enzyme from <i>Pseudomonas</i>, and also a homolog from <i>Streptomyces</i> ([https://parts.igem.org/Part:BBa_K5443014 BBa_K5443014]).
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This part builds on the work of earlier iGEM teams, who contributed ECH Parts as follows: [https://parts.igem.org/Part:BBa_K1129002 BBa_K1129002].
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<html>
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<figure>
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    <img src="https://static.igem.wiki/teams/5443/ech-reaction.jpg" style="width:600px;height:auto">
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    <figcaption>
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        <b>Figure 1. ECH-Ppu-catalysed conversion of feruloyl-CoA to vanillin.</b><br>
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        <i>This reaction represents the final step in our vanillin synthesis pathway. The ECH-Ppu enzyme converts feruloyl-CoA into vanillin.</i>
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    </figcaption>
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</figure>
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</html>

Latest revision as of 12:40, 2 October 2024

The Enoyl-CoA Hydratase/Aldolase (ECH) enzyme from Pseudomonas putida, ECH-Ppu, catalyses the fifth and final step in our vanillin biosynthesis pathway. ECH-Ppu converts feruloyl-CoA to vanillin by performing a two-step reaction: it hydrates the double bond of feruloyl-CoA, then cleaves the resulting thioester via a retro-aldol reaction (Fig. 1). The ECH enzyme can also potentially use caffeoyl-CoA and 4-coumaroyl-CoA as substrates.

We tested two alternate FCS enzymes for this pathway step; this enzyme from Pseudomonas, and also a homolog from Streptomyces (BBa_K5443014).

This part builds on the work of earlier iGEM teams, who contributed ECH Parts as follows: BBa_K1129002.

Figure 1. ECH-Ppu-catalysed conversion of feruloyl-CoA to vanillin.
This reaction represents the final step in our vanillin synthesis pathway. The ECH-Ppu enzyme converts feruloyl-CoA into vanillin.