Difference between revisions of "Part:BBa K200003"

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<partinfo>BBa_K200003 short</partinfo>
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<partinfo>BBa_K200003 short</partinfo><br>
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Rfal (Waal ligase) is an enzyme responsible for the ligation of an [http://en.wikipedia.org/wiki/O_antigen#O-antigen O-antigen] to the core [http://en.wikipedia.org/wiki/Oligosaccharide oligosaccharide] in the [http://en.wikipedia.org/wiki/Gram-negative_bacteria Gram-negative] bacterium's outer membrane.
  
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Unlike other exopolysaccharides, colanic acid does not naturally bind to the cell surface but rather forms a thick mesh between cells. While Waal Ligase usually links the O-antigen to the lipid A core, in K-12 it links colanic acid to the lipid A core. This is because E.coli K-12 does not produce O-antigen due to an insertion mutation.
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Unlike other exopolysaccharides, colanic acid does not naturally bind to the cell surface but rather forms a thick mesh between cells. While Waal Ligase usually links the O-antigen to the core oligosaccharide, in K-12 it links colanic acid to the core oligosaccharide. This is because E.coli K-12 does not produce O-antigen due to an insertion mutation.
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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Waal ligase is an enzyme responsible for the ligation of an O-antigen
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[[Image:II09_LPS.jpg|100px|frame|left|<i>Figure 1:</i> LPS structure (source: [http://en.wikipedia.org/wiki/O_antigen wikipedia])]]
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LipoPolySaccharyde (LPS) is a major component of the outer cell wall in Gram-negative bacteria. It is constituted of three main components: the O-antigen, the core polysaccharides and the lipid A (see Fig. 1).<br>
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<span class='h3bb'>Sequence and Features</span>
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The O-antigen is a highly variable chain of polysaccharydes and is physically linked to the core.<br>
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The core component is made up of a short chain of sugars (oligosaccharide) and is linked to both the O-antigen and the Lipid A components of the cell wall.<br>
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The Lipid A component of the LPS contains fatty acid chains which causes the LPS to dock into the outer membrane of the Gram-negative bacterium.<br>
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In the majority of E.coli, the enzyme Rfal joins the O-antigen to the membrane-bound lipid core molecule. Since the K-12 strain has an insertion mutation in the gene coding for O-antigen, the enzyme Rfal is free to join colanic acid to the lipid core.
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<br><br>The gene was used alongside [[Part:BBa_K200000 |RcsB]] and [[Part:BBa_K200002 |YgiV]] as part of the Imperial iGEM 2009 [http://2009.igem.org/Team:Imperial_College_London <i>The E.ncapsulator</i>] project to bind the colanic acid capsule to the cellular membrane.
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<br><br><br><br><br>
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<span class='h3bb'><b>Sequence and Features</b></span>
 
<partinfo>BBa_K200003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K200003 SequenceAndFeatures</partinfo>
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<br>
  
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===Requirements===
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*Gram-negative bacterium
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*Colanic acid producing gene ([[Part:BBa_K200000|rcsB]])
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
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#function pmid=15838026
 
#function pmid=15838026
 
#Function2 pmid=19019161
 
#Function2 pmid=19019161
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#Function3 pmid=15215252
 
</biblio>
 
</biblio>

Latest revision as of 16:17, 10 May 2013

Rfal
Rfal (Waal ligase) is an enzyme responsible for the ligation of an [http://en.wikipedia.org/wiki/O_antigen#O-antigen O-antigen] to the core [http://en.wikipedia.org/wiki/Oligosaccharide oligosaccharide] in the [http://en.wikipedia.org/wiki/Gram-negative_bacteria Gram-negative] bacterium's outer membrane.

Unlike other exopolysaccharides, colanic acid does not naturally bind to the cell surface but rather forms a thick mesh between cells. While Waal Ligase usually links the O-antigen to the core oligosaccharide, in K-12 it links colanic acid to the core oligosaccharide. This is because E.coli K-12 does not produce O-antigen due to an insertion mutation.

Usage and Biology

Figure 1: LPS structure (source: [http://en.wikipedia.org/wiki/O_antigen wikipedia])

LipoPolySaccharyde (LPS) is a major component of the outer cell wall in Gram-negative bacteria. It is constituted of three main components: the O-antigen, the core polysaccharides and the lipid A (see Fig. 1).
The O-antigen is a highly variable chain of polysaccharydes and is physically linked to the core.
The core component is made up of a short chain of sugars (oligosaccharide) and is linked to both the O-antigen and the Lipid A components of the cell wall.
The Lipid A component of the LPS contains fatty acid chains which causes the LPS to dock into the outer membrane of the Gram-negative bacterium.
In the majority of E.coli, the enzyme Rfal joins the O-antigen to the membrane-bound lipid core molecule. Since the K-12 strain has an insertion mutation in the gene coding for O-antigen, the enzyme Rfal is free to join colanic acid to the lipid core.

The gene was used alongside RcsB and YgiV as part of the Imperial iGEM 2009 [http://2009.igem.org/Team:Imperial_College_London The E.ncapsulator] project to bind the colanic acid capsule to the cellular membrane.




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 754
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Requirements

  • Gram-negative bacterium
  • Colanic acid producing gene (rcsB)


References

<biblio>

  1. function pmid=15838026
  2. Function2 pmid=19019161
  3. Function3 pmid=15215252

</biblio>