Wei Li,Chong Qiao,Jing Pang,Guolin Zhang,Yinggang Luo
Amaryllidaceae alkaloids are unique benzylphenethylamine derivatives that comprise of more than 600 members with a huge chemical diversity. Most of them showed interesting bioactivities, for instance, galanthamine (GAL) is clinically used for Alzheimer's disease treatment. All Amaryllidaceae alkaloids had been thought to be derived from 4′- O -methylnorbelladine originated from norbelladine catalyzed by norbelladine 4′- O -methyltransferase (N4OMT). Herein we mined the transcriptome datasets of Lycoris radiata , a GAL-producing plant. LrOMT was cloned, overexpressed in Escherichia coli , and purified to homogeneity. Bioinformatics analysis and enzymatic activity assays revealed that LrOMT is an S-adenosylmethionine-dependent Class I OMT. LrOMT exhibited both para - and meta - O -methylation activities toward norbelladine to give 4′- and 3′- O -methylnorbelladine. Twenty-four analogues, including the proposed biosynthetic intermediates, were introduced to investigate the substrate scope of LrOMT and it showed that the aromatic substrates should have two vicinal hydroxyl groups. The LrOMT-catalyzed O -methylation preference is dependent on the properties of the binding group of the substrates. The transcription levels of LrOMT were positively associated with the accumulation of the Amaryllidaceae alkaloids and the biosynthetic intermediates in L. radiata . The present work revealed that LrOMT catalyzes multiple O -methylation reactions and its characterization will be helpful to uncover novel biosynthetic genes for Amaryllidaceae alkaloids biosynthesis.
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