A microbial supply chain for production of the anti-cancer drug vinblastine

Abstract

Monoterpene indole alkaloids (MIAs) are a diverse family of complex plant secondary metabolites with many medicinal properties, including the essential anti-cancer therapeutics vinblastine and vincristine1. As MIAs are difficult to chemically synthesize, the world’s supply chain for vinblastine relies on low-yielding extraction and purification of the precursors vindoline and catharanthine from the plant Catharanthus roseus, which is then followed by simple in vitro chemical coupling and reduction to form vinblastine at an industrial scale2,3. Here, we demonstrate the de novo microbial biosynthesis of vindoline and catharanthine using a highly engineered yeast, and in vitro chemical coupling to vinblastine. The study showcases a very long biosynthetic pathway refactored into a microbial cell factory, including 30 enzymatic steps beyond the yeast native metabolites geranyl pyrophosphate and tryptophan to catharanthine and vindoline. In total, 56 genetic edits were performed, including expression of 34 heterologous genes from plants, as well as deletions, knock-downs and overexpression of ten yeast genes to improve precursor supplies towards de novo production of catharanthine and vindoline, from which semisynthesis to vinblastine occurs. As the vinblastine pathway is one of the longest MIA biosynthetic pathways, this study positions yeast as a scalable platform to produce more than 3,000 natural MIAs and a virtually infinite number of new-to-nature analogues.

Genome Assembly of the Medicinal Plant Voacanga thouarsii

“…Our genomic analysis showed that despite some similarities (e.g., similar gene content, absence of post-γ whole-genome duplication), V. thouarsii genome displays specific genomic features such as a higher TE content and a bigger size compared with other Apocynaceae . This new Apocynaceae genome thereby paves the way for a better understanding of MIA biosynthesis as well as the identification of new and/or more efficient MIA biosynthetic enzymes that can be used in the developing yeast cell factories producing MIAs ( Guirimand et al 2021 ; Kulagina et al 2021 ; Zhang et al 2022 ).…”

Biosynthesis Investigations of Terpenoid, Alkaloid, and Flavonoid Antimicrobial Agents Derived from Medicinal Plants

“…Using these approaches, genes of interest could be constructed in a highly effective way; meanwhile, the side pathways could be eliminated to a large extent. The targeted antimicrobial or resistance–reversal agents could be produced in the transgenic microorganisms or plants, which have had great success in the production of artemisinin [ 91 ], as well as other valuable compounds such as vinblastine [ 211 , 212 ], etoposide aglycone [ 213 ], vindoline, and catharanthine [ 212 ]. Lastly, with the continuous discovery of new phytochemicals, deep clarification of pharmacological mechanisms, and comprehensive understanding of specific biosynthesis pathways, plant-derived natural products will become increasingly more useful therapeutic antimicrobial candidates in the future.…”

MGCEP 1.0: A Genetic-Engineered Marine-Derived Chassis Cell for a Scaled Heterologous Expression Platform of Microbial Bioactive Metabolites

“…As a crucial part of the heterologous expression system, the efficiency and success rate are mostly determined by the chassis cells . There were some widely used hosts, including Escherichia coli, Saccharomyces cerevisiae, Pseudomonas putida, Aspergillus niger, Myxococcus xanthus, and Bacillus subtilis, developed for heterologous expression of primary or secondary metabolites. − Based on the important role in antibiotic production and relatively well-established genetic engineering systems, Streptomyces sp. has some advantages in the expression of Actinomycetes -derived BGCs compared with other bacteria and fungi, such as the abundant primary metabolic pathways, unique post-transcriptional modification system, and antibiotic resistance mechanism. ,, Streptomyces coelicolor A3(2) is a model bacterium for the study of Streptomyces sp.…”