Figure 2
From: An automated pipeline for the screening of diverse monoterpene synthase libraries
PinS domain organisation and location of the targeted amino acids. (a) Domain organisation of native PinS from Pinus Taeda (Uniprot ID: Q84KL6) and VAR3-PinS with indicated plasticity regions targeted in this study. The N-terminal domain of unknown function is shown in orange and the C-terminal class I terpene cyclase domain is shown in green. The amino acid residues in the three plasticity regions targeted in this study are indicated. Residues at positions 339 and 558 (in grey) were not targeted as they are highly conserved among plant mTC/S8. (b) Relative product profiles of native PinS and VAR3-PinS (data obtained from Leferink et al.8), the latter was used as template for further mutagenesis in this study. (c) The three targeted areas mapped onto a homology model of native PinS (SWISS-MODEL, Abies grandis α-bisabolene synthase (PDB: 3SAE40) was used as a template, which has 44.8% sequence identity with (-)-α-pinene synthase from Pinus taeda). The N-terminal domain is shown in orange, the C-terminal terpene cyclase domain in green, and the three targeted plasticity regions are highlighted in cyan sticks. The amino-acid sequences of recombinant native and VAR3-PinS are shown in Table S1 in the Supplementary Information available.
