Coding

Part:BBa_K4700001

Designed by: Thomas James Alexander   Group: iGEM23_BOKU-Vienna   (2023-10-12)


Delta24-Sterol Reductase (DHCR24)

Part type - Enzyme CDS - Class: Oxidoreductase (catalyzes e- transfer) - Coenzymes: (NADP+ / NADPH) or (NAD+ / NADH)

Other Names: Delta24-Sterol Reductase; Lanosterol Delta24-Reductase; EC: 1.3.1.72

Function: Reduction/Oxidation of double bond between the 24th and 25th carbon of sterane ring structure, on the B ring, of the sterol molecules suspended as in the cell membrane that serve as the substrate of our process. (e.x. Cholesterol [animals], Campesterol [plants], Ergosterol [yeasts])

In our anabolic process to ferment steroid hormones from simple carbon, this enzyme reduces the sterane ring to remove the double bond between C24/C25. This is a bond on the functional group of the central ring structure. This will likely not affect the function of the Cytochrome P450 enzymes too much as we use mammalian enzymes primarily and they are primarily sensitive to the sterane center.

In the Boku 2023 Process this enzyme is paired with DHCR7 (BBa_K4700000) for the purpose of engineering Cholesterol into yeast. (As well as knocking out the ERG5 and ERG6 Genes) Reactions Catalyzed:

Zymosterol -> Cholesta-8-en-3B-ol Cholesta-7,24-dien-3B-ol -> Lathosterol 7-Dehydrodesmosterol]] -> 7-Dehydrocholesterol Desmosterol -> Cholesterol

p1.jpg Figure (Restriction Digest) : BB2ABC + pGAP + 24DH + RPL2A

bokuhormownpathway.jpg Figure: Boku Vienna 2023 Complete Biosynthetic Pathway

Organism of Origin:

  -BOKU-Vienna 2023: Danio rerio (Zebra Fish)
  -(Souza et al, 2011): Bovine


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1439
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1439
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1439
    Illegal BglII site found at 1527
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1439
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1439
    Illegal AgeI site found at 512
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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