Figure 5.

Evolution of TⅡASs. (A) Identification of TⅡASs from Salvia species. CT and TⅡA are the substrate and product, respectively. B Phylogenetic tree of TⅡASs with reported 2-ODD activities. The phylogenetic tree was constructed by the maximum likelihood method, and the branches indicate bootstrap values calculated by the 1,000 permutaion test. Scale bar indicates 0.2 substitutions per position in the sequence. The classification was according to Kawai et al. ( 2014) and red fonts indicate the DOXC D4H/BX6 clade. C, Multiple sequence alignment of TⅡASs and other 2-ODD proteins with the conserved and specified domains highlighted. D, Steady-state kinetic parameters of SmTIIAS and its substitution mutants by in vitro enzymatic assays. Assays were carried out in triplicate. Data represent the mean ± se of values.