Generator

Part:BBa_K902080

Designed by: Lisa Oberding, Ali Honarmand   Group: iGEM12_Calgary   (2012-10-25)

PlacI/dszB/B0034/dszC

This part produces DszB and DszC under control of an inducible lacI promoter.

DszC enzymes have been shown to catalyze the oxidation of dibenzothiophene (DBT)to dibenzothiophene-5-oxide (DBTO) in the first reaction and then from DBTO to DBT sulfoxide (DBTO2) in the second reaction, both in the presence of NADH, oxygen, FMN and flavin reductases.

Ucalgary2012pathforDszC.png

HBPS desulfinase (DszB), which is a monomeric enzyme that carries out the last step of the 4S desulfurization pathway. This step carries out the conversion of 2-hydroxybiphenyl-2-sulfinic acid (HBPS) to 2-hydroxybiphenyl, releasing the sulfur in the form of sulfite. Naturally, this enzyme has the lowest activity of all the desulfurization genes involved in the 4S pathway, and thus this step is the rate limiting step. Because of this, the sequence has been mutated Y63F to increase enzyme activity.

Ucalgary2012 DszBpathway figure.PNG

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1428
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 652
    Illegal BamHI site found at 2232
    Illegal XhoI site found at 644
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 359
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2382


[edit]
Categories
Parameters
None