Structure and Biocatalytic Scope of Coclaurine N‐Methyltransferase. Issue 33 (28th June 2018)

• Record Type: Journal Article
• Authors: Bennett, Matthew R.; Thompson, Mark L.; Shepherd, Sarah A.; Dunstan, Mark S.; Herbert, Abigail J.; Smith, Duncan R. M.; Cronin, Victoria A.; Menon, Binuraj R. K.; Levy, Colin; Micklefield, Jason
• Is Part Of: Angewandte Chemie international edition. Volume 57:Issue 33(2018)
• Page Start: 10600
• Page End: 10604
• Publication Date: 2018-06-28
• Subjects: biocatalysis -- biosynthesis -- mechanism -- Methyltransferase -- structure; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201805060
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Benzylisoquinoline alkaloids (BIAs) are a structurally diverse family of plant secondary metabolites, which have been exploited to develop analgesics, antibiotics, antitumor agents, and other therapeutic agents. Biosynthesis of BIAs proceeds via a common pathway from tyrosine to ( S )‐reticulene at which point the pathway diverges. Coclaurine N ‐methyltransferase (CNMT) is a key enzyme in the pathway to ( S )‐reticulene, installing the N ‐methyl substituent that is essential for the bioactivity of many BIAs. In this paper, we describe the first crystal structure of CNMT which, along with mutagenesis studies, defines the enzymes active site architecture. The specificity of CNMT was also explored with a range of natural and synthetic substrates as well as co‐factor analogues. Knowledge from this study could be used to generate improved CNMT variants required to produce BIAs or synthetic derivatives. Abstract : Coclaurine N ‐methyltransferase (CNMT) installs the N ‐methyl substituent essential for the bioactivity of therapeutically important benzyl‐isoquinoline alkaloids (BIA). CNMT′s active site architecture is defined through structure guided mutagenesis and its biocatalytic scope is explored with synthetic substrates and co‐factor analogues. These studies provide insights for engineering and exploiting CNMT in the production of bioactive BIA and synthetic derivatives.