Translational_Unit
tphB

Part:BBa_K4378000

Designed by: Leander Maximilian Linke   Group: iGEM22_Uppsala   (2022-09-28)
Revision as of 14:03, 28 September 2022 by LeeAnd (Talk | contribs)


TphB: An effective overexpression casette

The first documented degradation of phthalates in soil bacteria comes from a paper published 1995. Researchers sampled sediment from the riverbed of the Passaic River in New Jersey (Wang, 1995). Samples were then exposed to various phthalates and their respective microbial populations and metabolites characterized thereafter. C. testosteroni emerged from a sample with a microbiota capable of growing on two different phthalate isomers. These were terephthalate (Tph), isophthalate, and p-hydroxybenzoate.

The designation of the gene TphB is used interchangeably with the name of its product’s function, decarboxylating cis-dihydrodiol dehydrogenase (DCDDH). TphB is unique in its capacity to decarboxylate the reaction product from the TphA1, A2, and A3 actions.

Bains et al. structurally characterized the 3-dimensional structure of DCDDH at 1.85å (Bains, 2012). The method used for structural characterization was iodide single wavelength anomalous dispersion. Computational modeling yielded information about how DCDDH acts on its substrate (Bains, 2012).

No genetic engineering has been carried out to improve catalytic activity of DCDDH. All published efforts outline characterizations of either mechanistic or structural interest. Similarly, little information exists as to the kinetics of the above catalysis accomplished by TphB. A comprehensive report on kinetics of phthalate ester metabolism is available from Kluwe et al. (Kluwe, 1982). Apart from the publication from Bains, no experiment has to date provided insight into how TphB accomplishes its reaction.


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 288
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 409


[edit]
Categories
//cds/transcriptionalregulator/repressor
//chassis/prokaryote/ecoli
//proteindomain/affinity
Parameters
chassisE. coli