Translational_Unit

Part:BBa_K1362020

Designed by: Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha   Group: iGEM14_Heidelberg   (2014-10-08)
Revision as of 21:32, 17 October 2014 by Cahlmann (Talk | contribs)

RBS + Xylanase

Xylanases are enzymes that degrad Xylan, a major component of wood. This is a modified version of BBa_K1175005 that is from B. subtilis (XylA). To ensure that it is kept intracellular we removed a signaling peptide from the N-terminus. Xylanases are interesting enzymes for industry as they have a huge variety of applications.


Usage and Biology

To characterize our part in respect to its thermal stability we conducted a Heatshock Assay. Firstly the Xylanases were incubated at 37 °C, 50 °C, 56 °C, 60 °C and 66 °C. After 30 minutes substrate (EnzChek® Ultra Xylanase Assay Kit) was added and the fluorescence was measured for 150 minutes in a platereader at 37 °C. The results indicate a reduction of activity at temperatures higher than 50 °C and complete loss of function for temperatures higher than 60 °C.

Activity after Heatshock assay for 30 minutes of linear Xylanase

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 22
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 426
    Illegal SapI.rc site found at 480


[edit]
Categories
Parameters
None