Part:BBa_K1216006

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 [http://www.rcsb.org/pdb/explore/explore.do?structureId=4KRX 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
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 951
  

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.[http://www.ncbi.nlm.nih.gov/pubmed/20978973]
2. [http://homepage.univie.ac.at/nikos.pinotsis/tev_protease.html University of Vienna TEV Protease info]
3. [http://www.cpcbiotech.it/EN/c/d/enzyme-portfolio/enzymes/acetyl-esterase-lyophilized CPC Biotech]
4. [http://www.ecocyc.org/new-image?type=GENE&object=EG11101 Ecocyc]
5. [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=ENZYME&object=MONOMER0-158&orgids=ECOLI Ecocyc]