Part:BBa_K2271116

ADH PTS1

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 880
  Illegal BamHI site found at 2047
  Illegal XhoI site found at 2083
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2433
  
• 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 additionally holds a PTS1 sequence for peroxisomal PEX5 import. For cytosolic expression please choose part: BBa_K2271115

Characterization

We verified the expression of ADH PTS1 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 PTS1 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