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Coding
BFRM52H

Part:BBa_K1438001

Designed by: Johann Bauerfeind   Group: iGEM14_Berlin   (2014-10-06)

Bacterioferritin (BFR) M52H heme-deletion

Optimized bacterioferritin and its properties

Iron Storaging Protein

Bacterioferritins are the E. coli cells natural iron storage proteins. These hollow nearly spherical protein shells detoxify the cell by sequestering excessive iron and forming Iron(III)hydroxid-oxide particels.

Bacterioferritin is an haem containing bacterial ferritin. Each heme is bound in a pocked formed by the interface between a pair of symmetry-related subunits [1]. However, it was investigated that these heme groups may be involved in the release of iron out of the ferritin iron core by forming an heme-mediated electron transfer to reduce immobilized Fe3+ to more soluble Fe2+.

We isolated this particular bacterioferritin from the probiotic strain E. coli Nissle 1917, which does have more iron aquisition systems than other well established E. coli strains. We conducted a site-directed mutagenesis to produce a haem free bacterioferritin protein [3].


Bacterioferritin as a mediator for magnetism in a cell

Bacterioferritins have been investigated regarding their magnetic character since the 80s. Bfr overexpressing E. coli Nissle 1917 - a strain with a low immunogenity- were previously used in studies as biological MRI contrast agents. [4] Furthermore, the magentic character of iron loading bacterioferritins were studied by Hawkins & Williams concluding that the contamination of the bacterioferritin iron core with phosphate reduces the super paramagnetic properties significantally. [5]

Usage and Biology

Improving BFRs Iron Storage Properties=

We constructed a improved version of this protein by site-directed Mutagenesis (M52H) and added it to the parts registry as BBa_K1438001.


Furthermore we constructed the iGEM Ferritin library which consists of 6 different ferritin protein expression devices, which can be used by future generations of iGEM students to further works with ferritins as nanocages. All of these parts are featured with an his-tag so purification and invitro studies may be conducted.

Quantification of Bacterioferritin mediated iron capacity increase

Expression of bacterioferritin increased the cells capacity to store iron

More iron can be stored inside of a cell that is overexpressing bacterioferritin.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 106


[edit]
Categories
//binding/metal
//cds
Parameters
protein19,7 kDa