Search bioRxiv⌕ Search

Biology subjects

Grogan, G.

Publications and source records attributed to Grogan, G..

2 recordsLinked to original sources

Biosynthesis of Securinega alkaloids via a cyanogenic glucoside-like pathway

The elucidation of biosynthetic pathways towards bioactive compounds in plants is essential for building biotechnological platforms for their scalable synthesis. Securinine and allosecurinine form part of the complement of Securinega alkaloids in Flueggea suffruticosa, a fundamental herb of Chinese medicine. Despite their potent biological activities and a growing interest in derivatives as pharmaceutical leads, key steps in their biosynthesis remain obscure. Here we report the discovery and reconstitution of the pathway to the core Securinega alkaloid precursor menisdaurilide. Using transcriptomics, Nicotiana benthamiana transient expression, in vitro assays, and pathway reconstitution in Saccharomyces cerevisiae, we reveal that three cytochrome P450s and a reductase are required for the biosynthesis of menisdaurilide from L-tyrosine via the cyano-metabolite menisdaurigenin, revealing a surprising convergence with the biosynthesis of cyanogenic glucosides such as dhurrin. The de novo biosynthesis of allosecurinine in N. benthamiana highlights the potential for preparative-scale heterologous production of Securinega alkaloids.

biochemistry↗

Nicotine biosynthesis completed by cryptic activating glucosylation

Nicotine is a neuroactive alkaloid produced by tobacco (Nicotiana tabacum) as a defense against herbivory, and an addictive stimulant that has been used by humans for millennia. Despite its significance, the core steps of its biosynthesis have remained elusive. Here, we demonstrate in vitro reconstruction of nicotine synthase, a four-enzyme stereoselective biocatalytic cascade that forms (S)-nicotine from nicotinic acid and N-methylpyrrolinium. This cascade includes two glucose-processing enzymes that participate in a cryptic activating glucosylation step. We also reconstruct this pathway in planta and present high resolution X-ray structures of the key oxidoreductases A622 and BBL bound to their substrate and product, respectively. This work establishes the complete biosynthetic pathway to nicotine, providing new gene targets for controlling alkaloid production in Nicotiana and unlocking enzymatic routes to pyridine alkaloids.

biochemistry↗