Coding

Part:BBa_K316003

Designed by: IC 2010 Team   Group: iGEM10_Imperial_College_London   (2010-10-22)
Revision as of 13:36, 22 October 2010 by Ks907 (Talk | contribs)

BBa_S04510

XylE - catechol 2,3-dioxygenase from P.putida with terminator

Catechol or catechol 2,3-dioxygenases + O(2) is converted by a ring cleavage into 2-hydroxymuconate semialdehyde which is the toxic and bright yellow-coloured substrate. This is a key enzyme in many (soil) bacterial species used for the degradation of aromatic compounds. The enzyme used here itself originating from Pseudomonas putida is a homotetramer of C230 monomers. The tetramerization interactions position a ferrous ion critical for enzymatic activity. It has been deduced that intersubunit interaction is essential to produce a functioning enzyme after performing N and C terminal modifications on the monomer. Coming together the subunits generate an active site. The reaction itself takes place within seconds after the addition by Pasteur pipette or spraying of catechol at a 100mM stock solution diluted with DDH20 (used by our lab.) The toxic byproduct is thought to interfere with cell wall integrity and cellular machinery such that exposed cells gradually die. 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 311
    Illegal NgoMIV site found at 483
    Illegal AgeI site found at 834
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
n/aXylE - catechol 2,3-dioxygenase from P.putida with terminator