CHE.496/2009/Planning

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==Arsenic sequestration==
==Arsenic sequestration==
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===Strategies===
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*Knockout arsA or arsB
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**Existing mutant strain: [http://cgsc2.biology.yale.edu/Mutation.php?ID=100082 arsB758(del)::kan]
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**Uncertain whether function is KO
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*Upregulate E. coli arsC (arsenate reductase)
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**Include copy of the gene on a plasmid, express at rates higher than wt
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*Transfer exogenous arsenate reductases to E.coli
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**Two families besides the E.coli arsC exist
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**Question of whether they are more efficient, require co-enzymes, etc
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*Express metal binding peptide in periplasm
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**Cys-Gly-Cys-Cys-Gly repeat + fusion to maltose binding protein for localization
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**[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=106604 Development of Bacterium-Based Heavy Metal Biosorbents: Enhanced Uptake of Cadmium and Mercury by Escherichia coli Expressing a Metal Binding Motif]
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===Pathway===
===Pathway===
<html><img src="http://www3.interscience.wiley.com/cgi-bin/fulltext/118914921/nf3"></html>
<html><img src="http://www3.interscience.wiley.com/cgi-bin/fulltext/118914921/nf3"></html>
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===Genes===
===Genes===
*arsA  
*arsA  

Revision as of 16:47, 17 May 2009

Arsenic sequestration

Strategies


Pathway

Genes

  • arsA
  • arsB
    • arsenite/antimonite transporter [ Escherichia coli str. K-12 substr. DH10B ]
    • GeneID: 6062297
  • arsC
    • arsenate reductase [ Escherichia coli str. K-12 substr. DH10B ]
    • GeneID: 6058451
    • Protein: Arsenate Reductase (ArsC) family, ArsC subfamily; arsenic reductases similar to that encoded by arsC on the R733 plasmid of Escherichia coli. E. coli ArsC catalyzes the reduction of arsenate [As(V)] to arsenite [As(III)], the first step in the detoxification of arsenic, using reducing equivalents derived from glutathione (GSH) via glutaredoxin (GRX). ArsC contains a single catalytic cysteine, within a thioredoxin fold, that forms a covalent thiolate-As(V) intermediate, which is reduced by GRX through a mixed GSH-arsenate intermediate. This family of predominantly bacterial enzymes is unrelated to two other families of arsenate reductases which show similarity to low-molecular-weight acid phosphatases and phosphotyrosyl phosphatases.

References

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