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1. Lisowsky, T. (1996). Removal of an intron with unique 3′ branch site creates an amino‐terminal protein sequence directing the scERV1 gene product to mitochondria. Yeast, 12(15), 1501-1510.
 
1. Lisowsky, T. (1996). Removal of an intron with unique 3′ branch site creates an amino‐terminal protein sequence directing the scERV1 gene product to mitochondria. Yeast, 12(15), 1501-1510.
 
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4. Hatahet, F., Nguyen, V. D., Salo, K. E., & Ruddock, L. W. (2010). Disruption of reducing pathways is not essential for efficient disulfide bond formation in the cytoplasm of E. coli. Microbial cell factories, 9(1), 67.
 
4. Hatahet, F., Nguyen, V. D., Salo, K. E., & Ruddock, L. W. (2010). Disruption of reducing pathways is not essential for efficient disulfide bond formation in the cytoplasm of E. coli. Microbial cell factories, 9(1), 67.
 
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Revision as of 22:35, 15 September 2019


Truncated Saccharomyces cerevisiae FAD-linked sulfhydryl oxidase Erv1p

The ERV1 gene from Saccharomyces cerevisiae encodes for a 189 amino acids protein, Erv1p, that is involved in different processes of mitochondrial biogenesis and maintenance1, 2. The protein shows a flavin-linked sulfhydryl oxidase enzymatic activity linked to the carboxy-terminal domain. Thus, Erv1p is capable of oxidizing thiol groups in proteins and catalyzing disulfide bond formation. A 15 kDa truncated version of Erv1p, consisting of the 117 amino acid residues carboxy-terminal domain, shows a similar or improved sulfhydryl oxidase activity compared to the full length protein2.


For a successful expression in E. coli, it is indispensable to avoid interactions that result in the aggregation of folding intermediates. Disulfide bonds can be involved in the structural, catalytic and signaling roles of the protein, but the formation of disulfide bonds can have certain problems that can cause misfolding, aggregation and low yields during the production of the protein3. The usage of Erv1p has been proposed as a mechanism for the formation of disulfide bonds in recombinant proteins in prokaryotic expression systems. It has been shown that, thanks to its ability to form disulfide bonds de novo, co-expression of Erv1p allows the proper formation of disulfide bonded proteins in the cytoplasm of E. coli4.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 58
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

1. Lisowsky, T. (1996). Removal of an intron with unique 3′ branch site creates an amino‐terminal protein sequence directing the scERV1 gene product to mitochondria. Yeast, 12(15), 1501-1510.
2. Lee, J. E., Hofhaus, G., & Lisowsky, T. (2000). Erv1p from Saccharomyces cerevisiae is a FAD‐linked sulfhydryl oxidase. FEBS letters, 477(1-2), 62-66.
3. Veggiani, G., & de Marco, A. (2011). Improved quantitative and qualitative production of single-domain intrabodies mediated by the co-expression of Erv1p sulfhydryl oxidase. Protein expression and purification, 79(1), 111-114.
4. Hatahet, F., Nguyen, V. D., Salo, K. E., & Ruddock, L. W. (2010). Disruption of reducing pathways is not essential for efficient disulfide bond formation in the cytoplasm of E. coli. Microbial cell factories, 9(1), 67.