Fig. 5 in A single residue determines substrate preference in benzylisoquinoline alkaloid N-methyltransferases

Fig. 5. Sequence-based prediction and in vivo screening of putative CNMT and RNMT enzymes. (A) The identity of residue 204 (E, glutamic acid; G, glycine) was used to predict CNMT- or RNMT-like function, respectively. (B) Overall amino acid sequence identity does not distinguish CNMT- or RNMT-like enzymes from each other (Supplementary Dataset 5). (C) Formation of N-methylated products following supplementation of yeast cultures with (S)-coclaurine (white bars), (S)-N- methylcoclaurine (black bars) and (S)-reticuline (grey bars). For each substrate, the culture with the greatest mean product concentration normalized to optical density was set to 100% and all other values were scaled accordingly (30.7μM OD−1 (S)-N-methylcoclaurine, HCANMT1; 3.4μM OD−1 (S)-N,N-dimethylcoclaurine, SCANMT1; 2.0μM OD−1 (S)-tembetarine, SCANMT1). Error bars represent standard deviation of four replicate cultures. Western blot analysis of NMT expression is show in Supplementary Fig. 6.Published as part of Morris, Jeremy S., Yu, Lisa & Facchini, Peter J., 2020, A single residue determines substrate preference in benzylisoquinoline alkaloid N-methyltransferases, pp. 1-11 in Phytochemistry (112193) 170 on page 7, DOI: 10.1016/j.phytochem.2019.112193, http://zenodo.org/record/829291