Difference between revisions of "Part:BBa K1216002"

(→‎Chromogenic enzymatic reaction)
(→‎Chromogenic enzymatic reaction)
Line 26: Line 26:
 
==='''Chromogenic enzymatic reaction'''===
 
==='''Chromogenic enzymatic reaction'''===
 
To check the functionality of our reporter construct, we tested the functionality of the enzyme by using the following enzymatic reaction resulting in a violet color.
 
To check the functionality of our reporter construct, we tested the functionality of the enzyme by using the following enzymatic reaction resulting in a violet color.
−
[[Image:Aeshydro.png|600px|left|thumb|]]
+
[[Image:Aeshydro.png|600px|none|thumb|]]
 
This reaction was tested in liquid culture as well as on colonies.
 
This reaction was tested in liquid culture as well as on colonies.
  

Revision as of 19:13, 1 October 2013

Acetyl esterase (aes) from Escherichia Coli

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]

A form of this protein with added 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[1] . Further it can be used for desacteylation of β-Lactam antibiotics[1] .


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[2] [3].


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


Chromogenic enzymatic reaction

To check the functionality of our reporter construct, we tested the functionality of the enzyme by using the following enzymatic reaction resulting in a violet color.

Aeshydro.png

This reaction was tested in liquid culture as well as on colonies.

Michaelis Menten kinetic of the Cell free extract with aes

AES reaction.png
Hydrolase AES.png

References

  1. [http://www.cpcbiotech.it/EN/c/d/enzyme-portfolio/enzymes/acetyl-esterase-lyophilized CPC Biotech]
  2. [http://www.ecocyc.org/new-image?type=GENE&object=EG11101 Ecocyc]
  3. [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=ENZYME&object=MONOMER0-158&orgids=ECOLI Ecocyc]