Difference between revisions of "Part:BBa K316003"
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<partinfo>BBa_K316003 short</partinfo>
 
<partinfo>BBa_K316003 short</partinfo>
  
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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.
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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. Catechol 2,3-dioxygenase (pdb id: 1MPY[http://www.ebi.ac.uk/pdbsum/1mpy]) was originally isolated from Pseudomonas putida and 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.
  
 
Please see ‘Part Design’ section for design considerations and parts used.
 
Please see ‘Part Design’ section for design considerations and parts used.
 +
 +
References:
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PubMed id: 10368270]
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http://www.ebi.ac.uk/pdbsum/1mpy
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Biochem. J. (2003) 371, 557–564 (Printed in Great Britain) 557
 +
Intersubunit interaction and catalytic activity of catechol 2,3-dioxygenases
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Akiko OKUTA1, Kouhei OHNISHI2,3, Sakiko YAGAME and Shigeaki HARAYAMA
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Marine Biotechnology Institute, 3-75-1 Heita, Kamaishi, Iwate 026-0001, Japan
 +
 +
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</biblio>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 15:46, 23 October 2010

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. Catechol 2,3-dioxygenase (pdb id: 1MPY[http://www.ebi.ac.uk/pdbsum/1mpy]) was originally isolated from Pseudomonas putida and 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.

Please see ‘Part Design’ section for design considerations and parts used.

References: PubMed id: 10368270] http://www.ebi.ac.uk/pdbsum/1mpy Biochem. J. (2003) 371, 557–564 (Printed in Great Britain) 557 Intersubunit interaction and catalytic activity of catechol 2,3-dioxygenases Akiko OKUTA1, Kouhei OHNISHI2,3, Sakiko YAGAME and Shigeaki HARAYAMA Marine Biotechnology Institute, 3-75-1 Heita, Kamaishi, Iwate 026-0001, Japan


</biblio>

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 337
    Illegal NgoMIV site found at 509
    Illegal AgeI site found at 860
  • 1000
    COMPATIBLE WITH RFC[1000]