Difference between revisions of "Part:BBa K1692002"
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[[File:SB2015_styrene_enzymes_SDS_PAGE.png|thumbnail|center|700px|This is a <b>SDS PAGE gel</b> with purified PAL, FDC and UbiX protein.  We ran a Mark 12 protein ladder to verify that our proteins were the correct molecular weight.]]<br><br>
 
[[File:SB2015_styrene_enzymes_SDS_PAGE.png|thumbnail|center|700px|This is a <b>SDS PAGE gel</b> with purified PAL, FDC and UbiX protein.  We ran a Mark 12 protein ladder to verify that our proteins were the correct molecular weight.]]<br><br>
  
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[[File:SB2015_styrene_absorbance.png|thumbnail|center|300px|<b>Styrene Absorbance</b>This is where we write]]<br><br>  
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[[File:SB2015_styrene_absorbance.png|thumbnail|center|400px|<b>Styrene Absorbance</b>The absorbance spectrum of pure styrene (1 mM) measured on a Spectramax Pro spectrophotometer]]<br><br>  
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[[File:SB2015_tCA_absorbance.png|thumbnail|center|300px|<b>Styrene Absorbance</b>This is where we write]]<br><br>  
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[[File:SB2015_tCA_absorbance.png|thumbnail|center|400px|<b>trans-Cinnamic Acid Absorbance</b>The absorbance spectrum of pure trans-Cinnamic Acid (1 mM) measured on a Spectramax Pro spectrophotometer]]<br><br>  
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[[File:SB2015_FMN_absorbance.png|thumbnail|right|300px|<b>Flavin Mononucleotide Absorbance</b>This is where we write]]<br><br>
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[[File:SB2015_FMN_absorbance.png|thumbnail|center|400px|<b>Flavin Mononucleotide Absorbance</b>The absorbance spectrum of pure Flavin Mononucleotide (1 mM) measured on a Spectramax Pro spectrophotometer]]<br><br>
  
 
<h2>Reference</h2>
 
<h2>Reference</h2>

Revision as of 23:52, 17 September 2015

codon optimized FDC with T7 promoter and Flag Tag

Introduction

Ferulic acid decarboxylase (FDC) catalyzes the conversion of trans-cinnamic acid to styrene. We codon-optimized S. cerevisiae’s FDC gene for expression in E. coli.

Styrene synthesis pathway The enzymes of interest are phenylalanine ammonia lyase (PAL), ferulic acid decarboxylase (FDC), and a flavin prenyltransferase involved in ubiquinone biosynthesis called UbiX. PAL catalyzes the conversion of phenylalanine to trans-cinnamic acid, while FDC catalyzes the conversion of trans-cinnamic acid to styrene. Recently, it has been discovered that a cofactor is required to activate FDC. This cofactor is a product of the reaction between dimethylallyl monophosphate (DMAP) and flavin mononucleotide (FMN), which is catalyzed by the enzyme UbiX.



Background

This is a SDS PAGE gel with purified PAL, FDC and UbiX protein. We ran a Mark 12 protein ladder to verify that our proteins were the correct molecular weight.


Styrene AbsorbanceThe absorbance spectrum of pure styrene (1 mM) measured on a Spectramax Pro spectrophotometer


trans-Cinnamic Acid AbsorbanceThe absorbance spectrum of pure trans-Cinnamic Acid (1 mM) measured on a Spectramax Pro spectrophotometer


Flavin Mononucleotide AbsorbanceThe absorbance spectrum of pure Flavin Mononucleotide (1 mM) measured on a Spectramax Pro spectrophotometer


Reference

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 233
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]