Regulatory

Part:BBa_K3784006

Designed by: LUXUAN LIU   Group: iGEM21_BNUZ-China   (2021-09-20)
Revision as of 15:14, 17 October 2021 by MANUL (Talk | contribs)


Fld


Fld consists of TAA1 (BBa_K3784004), fldH (BBa_K3784003), fldB (BBa_K3784001), fldC(BBa_K3784002) and acdA(BBa_K3784000), aiming to build a pathway in the engineered bacteria to metabolize tryptophan to produce indolepropionic acid, which can consume the remaining tryptophan and strengthen the intestinal mucosal barrier.


Usage and Biology

We reconstructed a five-step reaction. We cloned its tryptophan aminotransferase(TAA1) from Arabidopsis thaliana, which can catalyze the production of indole pyruvate from tryptophan. Then cloned from Clostridium sporogenes four enzymes -- fldB, fldC, fldH, and acdA in the fld gene cluster. fldB and fldC encode the 2-hydroxyacyl-CoA dehydratase protein family. fldH encodes phenyllactate dehydratase. acdA encodes 3-(aryl) acrylate reductase acdA. Tryptophan is first catalyzed by tryptophan aminotransferase(TAA1) to produce indolepyruvate, and then indolepyruvate is catalyzed by phenyllactate dehydratase (fldH) to produce indole lactic acid. This enzyme is also the rate-limiting enzyme of the entire reaction [2]. Then indole lactic acid is catalyzed by the 2-hydroxyacyl-CoA dehydratase protein family encoded by fldBC to produce indole acrylic acid, and finally indole acrylic acid is catalyzed by the 3-(aryl) acrylate reductase acdA encoded by acdA to produce indole propionic acid. In this way, our engineered bacteria can metabolize tryptophan to produce indolepropionic acid and repair the intestinal mucosal barrier

Characterization

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3649
    Illegal EcoRI site found at 3721
    Illegal XbaI site found at 4570
    Illegal SpeI site found at 5354
    Illegal SpeI site found at 5682
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3649
    Illegal EcoRI site found at 3721
    Illegal NheI site found at 1269
    Illegal SpeI site found at 5354
    Illegal SpeI site found at 5682
    Illegal NotI site found at 2374
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3649
    Illegal EcoRI site found at 3721
    Illegal BglII site found at 2528
    Illegal XhoI site found at 111
    Illegal XhoI site found at 1222
    Illegal XhoI site found at 6204
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3649
    Illegal EcoRI site found at 3721
    Illegal XbaI site found at 4570
    Illegal SpeI site found at 5354
    Illegal SpeI site found at 5682
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3649
    Illegal EcoRI site found at 3721
    Illegal XbaI site found at 4570
    Illegal SpeI site found at 5354
    Illegal SpeI site found at 5682
    Illegal AgeI site found at 385
  • 1000
    COMPATIBLE WITH RFC[1000]


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