Part:BBa_K4665001

Usage and Biology

This is the N-terminal of an ice nucleation protein which will be embedded into the outer cell membrane of E. coli. The sequence coding for the INPN is preceded by a pelB leader sequence. By attaching the pelB signal peptide in front of the INP protein, the fusion protein will be directed towards the bacterial periplasm where it will be anchored in the cell membrane (Singh et al., 2013). The INPN sequence is followed by two front-end sub-repeat sequences important for the stability of the fusion protein (Zhu et al., 2022).

INPN has been shown to be highly effective for whole cell catalysis and surface display, compared to other anchoring proteins such as Omp’s and Lpp (Hui et al., 2018).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 470
  Illegal PstI site found at 592
  Illegal NgoMIV site found at 54
  Illegal AgeI site found at 555
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

Hui, C. et al. (April 9, 2018). Surface display of PbrR on Escherichia coli and evaluation of the bioavailability of lead associated with engineering cells in mice. Scientific Reports 8(5685). https://doi.org/10.1038/s41598-018-24134-3

Singh, P., et al. (2013). Effect of signal peptide on stability and folding of Escherichia coli thioredoxin. PloS one, 8(5), e63442. https://doi.org/10.1371/journal.pone.0063442

Zhu, Y., et al. (December 6, 2021). Surface display of carbonic anhydrase on Escherichia coli for CO2 capture and mineralisation. Synthetic and Systems biotechnology, 7(1): 460-473. https://doi.org/10.1016%2Fj.synbio.2021.11.008