Figure 12.

Protein expression, sequence alignment, structure, and in vitro enzymatic analysis of γ-terpinene synthase TqTPS2 (Tq13G005250.1) from T. quinquecostatus. (A) Protein expression results for the control vector without γ-terpene synthase. 1, protein products after 0 hours of induction; 2, protein products after 12 hours of induction; 3, supernatant of purified protein; 4, sediment of purified protein; 5–8, eluant of purified protein (orange arrows indicate the eluant containing the target protein). (B) Protein expression results for the vector with TqTPS2. 1, supernatant of purified protein; 2, sediment of purified protein; 3, protein products after 0 hours of induction; 4, protein products after 12 hours of induction; 5–8, eluant of purified protein (arrows indicate the eluant containing the target protein). (C) Amino acid sequence alignment of TPS proteins from T. quinquecostatus, T. caespititius, and T. vulgaris. (D) 3D structure of the deduced TqTPS2 protein sequence. (E) In vitro enzymatic analysis of TqTPS2 using GPP as a substrate. Enzymatic products of TqTPS1 (green line) [22], enzymatic products of TqTPS2 (red line), products of authentic standard γ-terpinene (blue line), products of GPP (yellow line), and products of pGEX-4 T (control vector without terpene synthase insert) (purple line).