Difference between revisions of "Part:BBa K1450006"
 
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==Background and rational==
 
==Background and rational==
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Caulobacter crescentus is a bacterium frequently inhabiting fresh water lakes and streams. An important model for understanding microbial physiology, Caulobacter is known for it’s differentiated cell cycle. In particular, cells switch between two states; a motile “swarmer” cell and a static “stalked” cell. Here, the Caulobacter lifestyle provides a chasis for engineering solutions that require mobility interspersed with surface binding. Another potentially useful Caulobacter trait a regularly arrayed protein surface layer known as the S-layer. The S-layer is most frequently associated with protection, as the bacterium remains covered at all times throughout it’s lifestyle.  Indeed, a platform protein expression system has been engineered to exploit the S-layer for simple, scalable protein display. In recent years, there have been several examples where target proteins (such as immunoglobulins) are displayed to bind pathogen associated molecular protein (PAMPS).  
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''Caulobacter crescentus'' is a bacterium frequently inhabiting fresh water lakes and streams. An important model for understanding microbial physiology, ''Caulobacter'' is known for it’s differentiated cell cycle. In particular, cells switch between two states; a motile “swarmer” cell and a static “stalked” cell. Here, the ''Caulobacter'' lifestyle provides a chasis for engineering solutions that require mobility interspersed with surface binding. Another potentially useful ''Caulobacter'' trait a regularly arrayed protein surface layer known as the S-layer. The S-layer is most frequently associated with protection, as the bacterium remains covered at all times throughout it’s lifestyle.  Indeed, a platform protein expression system has been engineered to exploit the S-layer for simple, scalable protein display. In recent years, there have been several examples where target proteins (such as immunoglobulins) are displayed to bind pathogen associated molecular protein (PAMPS).  
  
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The Caulobacter traits above could be useful for applications in biomining, where a bacterium can be imagined to bind and recover select minerals from a complex mixture. Recently, several peptides were discovered that selectively bind to economically important minerals in ore slurries. We therefore hypothesized that these peptides could be integrated into the Caulobacter S-layer as a proof of concept for biomining applications.   
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The ''Caulobacter'' traits above could be useful for applications in biomining, where a bacterium can be imagined to bind and recover select minerals from a complex mixture. Recently, several peptides were discovered that selectively bind to economically important minerals in ore slurries. We therefore hypothesized that these peptides could be integrated into the ''Caulobacter'' S-layer as a proof of concept for biomining applications.   
  
 
==Experimental overview==
 
==Experimental overview==
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As a preliminary test, we observed increased Chalcopyrite precipitation with Caulobacter cells expressing a Chalcopyrite binding peptide in the S-layer. While non-quantitative, the data showed noticeable differences in precipitate settle when the peptide expressed in various locations in the S-layer. See the video below.  
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As a preliminary test, we observed increased Chalcopyrite precipitation with ''Caulobacter'' cells expressing a Chalcopyrite binding peptide in the S-layer. While non-quantitative, the data showed noticeable differences in precipitate settle when the peptide expressed in various locations in the S-layer. See the video below.  
  
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  <!-- source src="http://2014.igem.org/File:UBC_iGEM_chalcopyrite_biomining_2.mp4" -->
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  <source src="https://static.igem.org/mediawiki/2014/5/59/UBC_iGEM_chalcopyrite_biomining_2.mp4">
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</video>
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<center>
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'''Figure 1.''' Various peptides from Curtis ''et al.'' are displayed in different positions in the S-layer of ''Caulobacter'' cells. The cells were mixed with calcopyrite and precipitate settling was observed. 
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</center>
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==References==
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Curtis ''et al.'' 2008. Biomining with bacteriophage: Selectivity of displayed peptides for naturally occurring sphalerite and chalcopyrite. Biotechnology and bioengineering. '''102''':644-650.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 22:43, 31 October 2014

WSD-1 Chalcopyrite-binding peptide

WSD-1 Chalcopyrite-binding peptide

Background and rational

Caulobacter crescentus is a bacterium frequently inhabiting fresh water lakes and streams. An important model for understanding microbial physiology, Caulobacter is known for it’s differentiated cell cycle. In particular, cells switch between two states; a motile “swarmer” cell and a static “stalked” cell. Here, the Caulobacter lifestyle provides a chasis for engineering solutions that require mobility interspersed with surface binding. Another potentially useful Caulobacter trait a regularly arrayed protein surface layer known as the S-layer. The S-layer is most frequently associated with protection, as the bacterium remains covered at all times throughout it’s lifestyle. Indeed, a platform protein expression system has been engineered to exploit the S-layer for simple, scalable protein display. In recent years, there have been several examples where target proteins (such as immunoglobulins) are displayed to bind pathogen associated molecular protein (PAMPS).

The Caulobacter traits above could be useful for applications in biomining, where a bacterium can be imagined to bind and recover select minerals from a complex mixture. Recently, several peptides were discovered that selectively bind to economically important minerals in ore slurries. We therefore hypothesized that these peptides could be integrated into the Caulobacter S-layer as a proof of concept for biomining applications.

Experimental overview

As a preliminary test, we observed increased Chalcopyrite precipitation with Caulobacter cells expressing a Chalcopyrite binding peptide in the S-layer. While non-quantitative, the data showed noticeable differences in precipitate settle when the peptide expressed in various locations in the S-layer. See the video below.

Figure 1. Various peptides from Curtis et al. are displayed in different positions in the S-layer of Caulobacter cells. The cells were mixed with calcopyrite and precipitate settling was observed.

References

Curtis et al. 2008. Biomining with bacteriophage: Selectivity of displayed peptides for naturally occurring sphalerite and chalcopyrite. Biotechnology and bioengineering. 102:644-650.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]