Difference between revisions of "Part:BBa K1450006"
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==Background and rational== | ==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). | + | ''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. | + | 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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<center> | <center> | ||
<video width="400" controls> | <video width="400" controls> | ||
| − | <source src="http://2014.igem.org/File:UBC_iGEM_chalcopyrite_biomining_2.mp4"> | + | <!-- source src="http://2014.igem.org/File:UBC_iGEM_chalcopyrite_biomining_2.mp4" --> |
| + | <source src="https://static.igem.org/mediawiki/2014/5/59/UBC_iGEM_chalcopyrite_biomining_2.mp4"> | ||
Your browser does not support the video tag. | Your browser does not support the video tag. | ||
</video> | </video> | ||
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<center> | <center> | ||
| − | 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. | + | '''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. |
</center> | </center> | ||
| − | == | + | ==References== |
| − | Curtis ''et al.'' Biomining with bacteriophage: Selectivity of displayed peptides for naturally occurring sphalerite and chalcopyrite. Biotechnology and bioengineering. 102:644-650 | + | Curtis ''et al.'' 2008. Biomining with bacteriophage: Selectivity of displayed peptides for naturally occurring sphalerite and chalcopyrite. Biotechnology and bioengineering. '''102''':644-650. |
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]