Difference between revisions of "Part:BBa K4147006"
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<partinfo>BBa_K4147006 short</partinfo>
 
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The DsbA enzyme from <i>P. aeruginosa</i> is one of the most oxidizing proteins yet characterized [1]. This protein introduces disulfide bonds into proteins secreted to the periplasm. It was later optimized for <i>E. coli </i> strains.
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The DsbA enzyme from <i>P. aeruginosa</i> is one of the most oxidizing proteins yet characterized [1]. This protein introduces disulfide bonds into proteins secreted to the periplasm. It was later optimized for <i>E. coli </i> strains. </p>
  
 
===Usage and Biology===
 
===Usage and Biology===
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<p align="justify">
 
Fusing a recombinant protein's N-terminus to a signal peptide is one way to increase the solubility and functionality of the protein in <i>E. coli</i>. This can cause the heterologous protein to be exported from the cytoplasm into the periplasm or the culture media via the type II secretion system [1].  
 
Fusing a recombinant protein's N-terminus to a signal peptide is one way to increase the solubility and functionality of the protein in <i>E. coli</i>. This can cause the heterologous protein to be exported from the cytoplasm into the periplasm or the culture media via the type II secretion system [1].  
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One of the recognized proteins that oxidizes the most is DsbA. Although DsbA homologs from other bacteria, such as <i>Pseudomonas aeruginosa</i>, are even more oxidizing than <i>E. coli</i> DsbA, which has a redox potential of -119 mV (-95 mV). Numerous secreted polypeptides, including virulence factors, are folded oxidatively by DsbA. About 300 proteins in <i>E. coli</i>, or 40% of the periplasmic proteome, have an even number of cysteine residues and may therefore be folded by DsbA [2].
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One of the recognized proteins that oxidizes the most is DsbA. Although DsbA homologs from other bacteria, such as <i>Pseudomonas aeruginosa</i>, are even more oxidizing than <i>E. coli</i> DsbA, which has a redox potential of -119 mV (-95 mV). Numerous secreted polypeptides, including virulence factors, are folded oxidatively by DsbA. About 300 proteins in <i>E. coli</i>, or 40% of the periplasmic proteome, have an even number of cysteine residues and may therefore be folded by DsbA [2]. </p>
  
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Among the TRX-related proteins known to exist, DsbA is the principal disulfide bond introducer and has the second-highest redox potential in the periplasm of <i>E. coli</i>. DsbA was frequently used in the development of commercial expression plasmids to support periplasmic or extracellular secretion based on this property. Adding proteins to DsbA peptides can improve the target molecule's performance and solubility while also drastically reducing the creation of inclusion bodies [1].  
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<p align="justify">
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Among the TRX-related proteins known to exist, DsbA is the principal disulfide bond introducer and has the second-highest redox potential in the periplasm of <i>E. coli</i>. DsbA was frequently used in the development of commercial expression plasmids to support periplasmic or extracellular secretion based on this property. Adding proteins to DsbA peptides can improve the target molecule's performance and solubility while also drastically reducing the creation of inclusion bodies [1]. </p>
  
 
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Revision as of 06:05, 10 October 2022


Disulfide interchange protein DsbA from Pseudomonas aeruginosa

The DsbA enzyme from P. aeruginosa is one of the most oxidizing proteins yet characterized [1]. This protein introduces disulfide bonds into proteins secreted to the periplasm. It was later optimized for E. coli strains.

Usage and Biology

Fusing a recombinant protein's N-terminus to a signal peptide is one way to increase the solubility and functionality of the protein in E. coli. This can cause the heterologous protein to be exported from the cytoplasm into the periplasm or the culture media via the type II secretion system [1]. One of the recognized proteins that oxidizes the most is DsbA. Although DsbA homologs from other bacteria, such as Pseudomonas aeruginosa, are even more oxidizing than E. coli DsbA, which has a redox potential of -119 mV (-95 mV). Numerous secreted polypeptides, including virulence factors, are folded oxidatively by DsbA. About 300 proteins in E. coli, or 40% of the periplasmic proteome, have an even number of cysteine residues and may therefore be folded by DsbA [2].

Among the TRX-related proteins known to exist, DsbA is the principal disulfide bond introducer and has the second-highest redox potential in the periplasm of E. coli. DsbA was frequently used in the development of commercial expression plasmids to support periplasmic or extracellular secretion based on this property. Adding proteins to DsbA peptides can improve the target molecule's performance and solubility while also drastically reducing the creation of inclusion bodies [1].

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 219
    Illegal NgoMIV site found at 403
  • 1000
    COMPATIBLE WITH RFC[1000]


REFERENCES

[1] Zhang, W., Lu, J., Zhang, S., Liu, L., Pang, X., & Lv, J. (2018). Development an effective system to expression recombinant protein in E. coli via comparison and optimization of signal peptides: Expression of Pseudomonas fluorescens BJ-10 thermostable lipase as case study. Microbial Cell Factories, 17(1). doi:10.1186/s12934-018-0894-y

[2] Shouldice, S. R., Heras, B., Jarrott, R., Sharma, P., Scanlon, M. J., & Martin, J. L. (2010). Characterization of the DsbA Oxidative Folding Catalyst from Pseudomonas aeruginosa Reveals a Highly Oxidizing Protein that Binds Small Molecules. Antioxidants & Redox Signaling, 12(8), 921–931. doi:10.1089/ars.2009.2736