Coding
ADH

Part:BBa_K2271115

Designed by: iGEM Cologne-Düsseldorf   Group: iGEM17_Cologne-Duesseldorf   (2017-10-27)
Revision as of 18:53, 30 October 2017 by Dani94 (Talk | contribs)

ADH alcohol dehydrogenase

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal suffix found in sequence at 2047
    Illegal BglII site found at 880
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2402
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

We used an alcohol dehydrogenase(ADH) as part of our nootkatone pathway. The ADH we used for our project is from Pichia pastoris. It converts (+)-trans-nootkatol into (+)-nootkatone by oxidation, using NAD+ as a cofactor. A cofactor regeneration is not necessary in our case, because BM3, another enzyme of our nootkatone pathway, degrades NADH+H+ into NAD+. In nature ADH reduces or oxidizes alcohols to their specific aldehydes or ketones with help of NAD+.

This part is designed for cytosolic expression, for peroxisomal import please choose part: BBa_K2271116


Characterization

We verified the expression of ADH via western blot. Therefore, we have a 3xFLAG-6xHis-Tag as a part in our plasmids, we can use for the antibodies. The estimated atomic mass of ADH is 38kDa.


Protein abundance in WT and transformed cells from Saccharomyces cerevisiae: Protein abundance was detected using 6x His Tag Antibody. WT = wild type, ADH = Alcohol Dehydrogenase, PTS1 = Peroxisom Targeting Signal 1


References

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Categories
Parameters
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