Part:BBa_K808027

EstA : inactive fusion protein for bacterial surface expression

EstA is a membrane bound but inactive esterase from Pseudomonas aeruginosa. It can be used (unlike https://parts.igem.org/Part:BBa_K633001 BBa_K633001) for the cell surface expression of enzymes. Similiar to OmpA it allows to perform bacterial display of e.g. hydrolases like it has already been shown by Becker, Theile, Heppeler & Michalczyk, 2005. We used EstA to create our chimeric protein BBa_K808030 (Figure 1).

Figure 1:Visualization of BBa_K808030: EstA (red & grey) anchoring a second protein (pNB-Est13 in yellow) into a biological membrane

IISER_BHOPAL 2021

Cold adapted Esterase

A cold-adapted esterase was extracted from psychrotrophic bacterium Pseudoalteromonas sp. strain 643A, which is present in the alimentary tract of Antarctic krill Euphasia superba Dana. This protein consists of 206 amino acids residue with a molecular mass of 23,036 Da and belongs to the member of the GDSL-lipolytic enzymes family.1

References

Cieśliński, H., Białkowska, A. M., Długołęcka, A., Daroch, M., Tkaczuk, K. L., Kalinowska, H., Kur, J., & Turkiewicz, M. (2007). A cold-adapted esterase from psychrotrophic Pseudoalteromas sp. strain 643A. Archives of Microbiology, 188(1), 27–36. https://doi.org/10.1007/s00203-007-0220-2

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 97
  Illegal NgoMIV site found at 235
  Illegal NgoMIV site found at 775
  Illegal NgoMIV site found at 1144
  
• 1000
  COMPATIBLE WITH RFC[1000]