Generator

Part:BBa_K1216006

Designed by: Parvathi Chandran   Group: iGEM13_ETH_Zurich   (2013-08-29)

Acetyl esterase (aes) from Escherichia Coli with TEV and poly-HIS tags

Acetyl esterase - This is a cytosolic hydrolase that can catalyze hydrolysis of esters of p-nitrophenyl derivatives

3D representation of the acetyl esterase from RCSB
The poly-HIS tag can be used for protein purification (IMAC)[1]. The TEV tag can then be used to have the TEV protease specifically cleave off the poly-HIS tag from the purified protein [2].

A form of this protein without TEV and poly-HIS tags can be found here.

Usage and Biology

The hydrolase capacity of acetyl esterase makes it suitable for reporter application with substrates like acetylated xylan, ethyl acetate, cephalosporin C and derivatives[3] . Further it can be used for desacteylation of β-Lactam antibiotics[3] .


The acetyle esterase, apart from its hydrolase activity also binds competitively to malT, a transcription activator for the maltose operon, thus inhibiting regulation of genes required for maltose catabolism[4] [5].


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 27
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

The final construct was sequenced.

References

  1. Loghran ST, "Purification of poly-histidine-tagged proteins.",Methods Mol Biol. 2011;681:311-35. doi: 10.1007/978-1-60761-913-0_17.[1]
  2. University of Vienna TEV Protease info
  3. CPC Biotech
  4. Ecocyc
  5. Ecocyc
[edit]
Categories
Parameters
None