Difference between revisions of "Part:BBa K4428001"

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For the biosorption of lead ions, PbrR has been effectively engineered onto the surface of microorganisms. However, this molecule's lack of specificity makes it challenging to recover a single heavy metal ion selectively. The PbrR coming from the MerR family was characterized structurally by two distinct domains: an N-terminal DNA-binding domain (DBD) and a C-terminal metal binding domain (MBD) containing a central linker. Studies claimed that the MerR member was made up of the C-terminal conserved MBD and the N-terminal distinct DBD (Chang et al. 2015). It is known that the smaller the molecule the better the cell surface expression. Also, studies showed that both the domain’s functions are independent of each other (Song et al. 2004; Tao et al. 2013). This led to the expression of just the metal-binding-domain on the cell surface to increase the surface hence increasing the adsorption capacity.<br><br>
 
For the biosorption of lead ions, PbrR has been effectively engineered onto the surface of microorganisms. However, this molecule's lack of specificity makes it challenging to recover a single heavy metal ion selectively. The PbrR coming from the MerR family was characterized structurally by two distinct domains: an N-terminal DNA-binding domain (DBD) and a C-terminal metal binding domain (MBD) containing a central linker. Studies claimed that the MerR member was made up of the C-terminal conserved MBD and the N-terminal distinct DBD (Chang et al. 2015). It is known that the smaller the molecule the better the cell surface expression. Also, studies showed that both the domain’s functions are independent of each other (Song et al. 2004; Tao et al. 2013). This led to the expression of just the metal-binding-domain on the cell surface to increase the surface hence increasing the adsorption capacity.<br><br>
  
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Importantly, the Pb2+ adsorption capacity of PbrR MBD-displayed cells was about 1.92-fold higher than that of the full-length PbrR-displayed cells. Also, the PbrR MBD shows highly selective adsorption towards the lead similar to the full-length PbrR (Hui et al. 2018).  
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Importantly, the Pb<sup>2+</sup> adsorption capacity of PbrR MBD-displayed cells was about 1.92-fold higher than that of the full-length PbrR-displayed cells. Also, the PbrR MBD shows highly selective adsorption towards the lead similar to the full-length PbrR (Hui et al. 2018).
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[[File:IITD_PbrR_MBD_Structure.png|800px|thumb|centre|A model of PbrR protomer generated using Swiss-Model workspace (https://swissmodel.expasy.org/workspace/). The ribbon diagram shows the Pb<sup>2+</sup> binding domain in color and the other parts in gray. Residues Cys79, Cys114, and Cys123 involved in metal binding are presented as a space-filling model. d Ribbon representation of PbBD generated by truncating 49 N-terminal amino acids and 15 C-terminal amino acids of PbrR]]
  
 
==Part Uses==
 
==Part Uses==

Revision as of 21:28, 9 October 2022


PbrR MBD - Lead-binding domain of PbrR

PbrR was modified by truncating the N-terminal DNA Binding Domain and C-terminal redundant amino acid residues to obtain lead domain maintaining its Pb2+ binding property. The MBD (metal binding domain) being smaller in size leads to increased display and expression than the full-length PbrR-displayed cells.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 229
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 229
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 229
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 229
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 20


Background

For the biosorption of lead ions, PbrR has been effectively engineered onto the surface of microorganisms. However, this molecule's lack of specificity makes it challenging to recover a single heavy metal ion selectively. The PbrR coming from the MerR family was characterized structurally by two distinct domains: an N-terminal DNA-binding domain (DBD) and a C-terminal metal binding domain (MBD) containing a central linker. Studies claimed that the MerR member was made up of the C-terminal conserved MBD and the N-terminal distinct DBD (Chang et al. 2015). It is known that the smaller the molecule the better the cell surface expression. Also, studies showed that both the domain’s functions are independent of each other (Song et al. 2004; Tao et al. 2013). This led to the expression of just the metal-binding-domain on the cell surface to increase the surface hence increasing the adsorption capacity.

Importantly, the Pb2+ adsorption capacity of PbrR MBD-displayed cells was about 1.92-fold higher than that of the full-length PbrR-displayed cells. Also, the PbrR MBD shows highly selective adsorption towards the lead similar to the full-length PbrR (Hui et al. 2018).

A model of PbrR protomer generated using Swiss-Model workspace (https://swissmodel.expasy.org/workspace/). The ribbon diagram shows the Pb2+ binding domain in color and the other parts in gray. Residues Cys79, Cys114, and Cys123 involved in metal binding are presented as a space-filling model. d Ribbon representation of PbBD generated by truncating 49 N-terminal amino acids and 15 C-terminal amino acids of PbrR

Part Uses

In contrast to the conventional physical and chemical methods for environmental remediation, microbial cell-surface display has evolved as a unique strategy for the bioremediation of heavy metas. An innovative method for the selective bioremediation of harmful heavy metals is the engineering of the particular metal binding domain or motif derived from the MerR family regulators on the microbial cell surface.

The PbrR MBD domain due to its smaller size can lead to increased cell surface display causing better adsorption of the metal ion. Using the MBD instead of the full length PbrR for expression on cell surface will prove better for bioremediation of the heavy metal Pb2+ ions.