Difference between revisions of "Part:BBa K2662002"

Latest revision as of 06:49, 17 October 2018

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.

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

Results

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 how 7-epi-sesquithujene production varied with growing conditions, such as differing optical density at the introduction of IPTG and n-octane. The results show that the amount of sesquiterpene production can vary widely with growing conditions.

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