Part:BBa_K5357003

Pectate lyase B (PelB) Signal Sequence

Usage and Biology

PelB (pectate lyase B) is a 22 amino acid protein sequence that originates from the Erwinia carotovora pectate lyase B gene. It is a signal peptide that is important for the correct localisation of both membrane and secretory proteins. When fused to a recombinant protein in E. coli via the N-terminus, that protein will be targeted to the periplasmic space, where the PelB sequence will then be cleaved off.

The purpose of targeting proteins to the periplasm is to allow for proper disulphide bond formation. The cytoplasm is a reducing environment, favouring the reduction of S-S bonds to -SH HS- groups, whereas the periplasm is oxidizing, enabling the oxidation of -SH HS- to S-S (a disulphide bond) and water. This signal sequence therefore enables the correct folding of proteins that have disulphide bonds in E. coli.

Design Notes

The sequence was codon-optimised for expression in E.coli.

Source

http://www.signalpeptide.de/index.php?sess=&m=listspdb_bacteria&s=details&id=194499&listname=

References

Sockolosky JT, Szoka FC. Periplasmic production via the pET expression system of soluble, bioactive human growth hormone. Protein Expr Purif. 2013;87(2):129-35.

Lei SP, Lin HC, Wang SS, Callaway J, Wilcox G. Characterization of the Erwinia carotovora pelB gene and its product pectate lyase. J Bacteriol. 1987;169(9):4379-83.

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 54
  
• 1000
  COMPATIBLE WITH RFC[1000]