Coding

Part:BBa_K4350818

Designed by: Ryan Riopel   Group: iGEM22_UBC-Okanagan   (2022-10-09)

N. nambi Hipidin Synthase (nnHispS)

Hispidin synthase (HispS) is a polyketide synthase responsible for catalyzing the first step in the fungal bioluminescence pathway (Kotlobay et al., 2018). Polyketide synthases are a class of enzymes that produce secondary metabolites through the addition of malonyl moieties (alternating methyl and ketone groups) to carbon chains (Kotlobay et al., 2018). HispS catalyzes the synthesis of hispidin from a common fungal metabolite, caffeic acid. This reaction is facilitated through the addition of coenzyme A to caffeic acid, creating caffeyl-CoA and dephosphorylating ATP into AMP (Kotlobay et al., 2018). 2 molecules of malonyl-CoA are then utilized to add malonyl subunits to caffeyl-CoA, the molecule forms a heterocyclic ring, resulting in hispidin (Kotlobay et al., 2018).

Hispidin synthase is composed of several domains, an acyl-CoA synthetase region, a phosphopantetheine attachment site, and an acyl transferase domain (Kotlobay et al., 2018). Additionally, proper functioning of HispS requires the addition of a phosphopantetheinyl group to a conserved serine residue by a phosphopantetheinyl transferase enzyme (Kotlobay et al., 2018).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 63
    Illegal SpeI site found at 1194
    Illegal PstI site found at 904
    Illegal PstI site found at 3173
    Illegal PstI site found at 3926
    Illegal PstI site found at 4142
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1194
    Illegal PstI site found at 904
    Illegal PstI site found at 3173
    Illegal PstI site found at 3926
    Illegal PstI site found at 4142
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1944
    Illegal BglII site found at 4917
    Illegal BamHI site found at 1380
    Illegal BamHI site found at 2089
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 63
    Illegal SpeI site found at 1194
    Illegal PstI site found at 904
    Illegal PstI site found at 3173
    Illegal PstI site found at 3926
    Illegal PstI site found at 4142
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 63
    Illegal SpeI site found at 1194
    Illegal PstI site found at 904
    Illegal PstI site found at 3173
    Illegal PstI site found at 3926
    Illegal PstI site found at 4142
    Illegal NgoMIV site found at 3097
    Illegal AgeI site found at 1295
    Illegal AgeI site found at 3752
    Illegal AgeI site found at 4435
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  • Kotlobay, A. A., Sarkisyan, K. S., Mokrushina, Y. A., Marcet-Houben, M., Serebrovskaya, E. O., Markina, N. M., Gonzalez Somermeyer, L., Gorokhovatsky, A. Y., Vvedensky, A., Purtov, K. V., Petushkov, V. N., Rodionova, N. S., Chepurnyh, T. V., Fakhranurova, L. I., Guglya, E. B., Ziganshin, R., Tsarkova, A. S., Kaskova, Z. M., Shender, V., … Yampolsky, I. V. (2018). Genetically encodable bioluminescent system from fungi. Proceedings of the National Academy of Sciences, 115(50), 12728–12732. https://doi.org/10.1073/pnas.1803615115
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