Difference between revisions of "Part:BBa K316003"
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<partinfo>BBa_K316003 short</partinfo> | <partinfo>BBa_K316003 short</partinfo> | ||
| − | 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. | + | 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<cite>1</cite>. This is a key enzyme in many (soil) bacterial species used for the degradation of aromatic compounds. Catechol 2,3-dioxygenase (pdb id: 1MPY<cite>2</cite>[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. | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
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<!-- --> | <!-- --> | ||
| − | <span class='h3bb'>Sequence and Features</span> | + | <span class='h3bb'><big>'''Sequence and Features'''</big></span> |
| − | <partinfo> | + | |
| + | <partinfo>BBa_K316004 SequenceAndFeatures</partinfo> | ||
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
| − | <partinfo> | + | <partinfo>BBa_K316004 parameters</partinfo> |
<!-- --> | <!-- --> | ||
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| + | <span class='h3bb'><big>'''Part Characterisation'''</big></span> | ||
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| + | Characterisation data still to come | ||
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| + | ===References=== | ||
| + | <biblio> | ||
| + | #1 PubMed id: 10368270 | ||
| + | #2 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> | ||
Revision as of 16:07, 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 substrate1. This is a key enzyme in many (soil) bacterial species used for the degradation of aromatic compounds. Catechol 2,3-dioxygenase (pdb id: 1MPY2[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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 380
Illegal NgoMIV site found at 552
Illegal AgeI site found at 903 - 1000COMPATIBLE WITH RFC[1000]
Part Characterisation
Characterisation data still to come
References
<biblio>
- 1 PubMed id: 10368270
- 2 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>