Part:BBa_K2662002

7-Epi-Sesquithujene Synthase

This gene encodes for 7-epi-sesquithujene synthase, which is derived from Terpene Synthase 4 from Zea mays cultivar B73. It catalyzes farnesyl pyrophosphate to 7-epi-sesquithujene and other sesquiterpenoids, including beta-bisabolene.

Results

File:T--UNebraska-Lincoln--results fig 11 2018.png

Figure 1:Production of 7-epi-sesquithujene under differing growing conditions.

Codon-optimized 7-epi-sesquithujene synthase (TPS4CO) was paired with genes encoding the mevalonate-dependent pathway, which produces farnesyl pyrophosphate, in a two plasmid system. The cultures were overlaid with n-octane to partition the sesqueterpenoid products. Octane samples were analyzed through gas chromatography-mass spectrometry to identify and quantify sesqueterpenoid products. The adjacent results show that sesquiterpene production can vary widely with growing conditions, as exampled by the major product 7-epi-sesquithujene.

File:T--UNebraska-Lincoln--results fig 14 2018.png

Figure 2: Production comparison of 7-epi-sesquithujene to beta-bisabolene.

Six biological replicates were tested to determine if product production was consistent under identical growing conditions. For these replicates, 7-epi-sesquithujene and beta-bisabolene production were quantified. The adjacent results show that sesquiterpenoid production was greatly varied, with observed concentration of the major product ranging between 0.52 to 92.78 mg/L. There must be a variable controlling production that has not been identified yet. In these replicates, it was observed that although production varied, 7-epi-sesquithujene is the major product and beta-bisabolene is the second major product of 7-epi-sesquithujene synthase.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 58
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]