Coding
tphB

Part:BBa_K808010

Designed by: Sascha Hein, Sven Rumpf, Daniel Sachs   Group: iGEM12_TU_Darmstadt   (2012-09-02)
Revision as of 00:36, 27 September 2012 by DSachs (Talk | contribs) (→‎Part Characterisation)

tphB: reaction from DCD to protocatechuate

Homology modelling of TphB. For simulation parameters http://2012.igem.org/Team:TU_Darmstadt/Modeling_Homologie_Modeling#TphB click here.

TphB is coding for the decarboxylating cis-dihydrodiol dehydrogenase from Comamonas testosteroni Kf-1. It catalyzes the second reaction in the terephthalic acid degration - the decarboxylation of DCD to protocatechuate. The [http://2012.igem.org/Team:TU_Darmstadt/Modeling_Homologie_Modeling#TphB homology modeling] suggests a homodimerization as described by the recent paper by Bains et al.
For further information, please visit the [http://2012.igem.org/Team:TU_Darmstadt/Labjournal/Metabolism TU Darmstadt iGEM team wiki].











Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 155
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 202
    Illegal NgoMIV site found at 877
    Illegal AgeI site found at 939
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 76



Part Characterisation

Figure 1. Note the peak in FPLC at about 32 minutes (about 64 kDa)


FPLC confirmed the suggested homodimerization (Figure 1).

References

  • Sasoh, M., E. Masai, et al. (2006). "Characterization of the terephthalate degradation genes of Comamonas sp. strain E6." Appl Environ Microbiol 72(3): 1825-1832.
  • Fukuhara, Y., K. Inakazu, et al. (2010). "Characterization of the isophthalate degradation genes of Comamonas sp. strain E6." Appl Environ Microbiol 76(2): 519-527.
  • Kamimura, N., T. Aoyama, et al. (2010). "Characterization of the protocatechuate 4,5-cleavage pathway operon in Comamonas sp. strain E6 and discovery of a novel pathway gene." Appl Environ Microbiol 76(24): 8093-8101.
  • Schläfli HR, Weiss MA, Leisinger T, Cook AM. (1994)"Terephthalate 1,2-dioxygenase system from Comamonas testosteroni T-2: purification and some properties of the oxygenase component." J Bacteriol. 1994 Nov;176(21):6644-52.
  • Bains, J., J.E. Wulff, and M.J. Boulanger, Investigating Terephthalate Biodegradation: Structural Characterization of a Putative Decarboxylating cis-Dihydrodiol Dehydrogenase. J Mol Biol, 2012.
[edit]
Categories
//cds/biosynthesis
//function/degradation
Parameters
protein