A microbial supply chain for production of the anti-cancer drug vinblastine
Jie Zhang
(1)
,
Lea Hansen
(1)
,
Olga Gudich
(1)
,
Konrad Viehrig
(1)
,
Lærke Lassen
(1)
,
Lars Schrübbers
(1)
,
Khem Adhikari
(1)
,
Paulina Rubaszka
(1)
,
Elena Carrasquer-Alvarez
(1)
,
Ling Chen
(1)
,
Vasil D’ambrosio
(1)
,
Beata Lehka
(1)
,
Ahmad Haidar
(1)
,
Saranya Nallapareddy
(1)
,
Konstantina Giannakou
(1)
,
Marcos Laloux
(1)
,
Dushica Arsovska
(1)
,
Marcus Jørgensen
(1)
,
Leanne Jade G. Chan
(2)
,
Mette Kristensen
(1)
,
Hanne Christensen
(1)
,
Suresh Sudarsan
(1)
,
Emily Stander
(3)
,
Edward Baidoo
(2)
,
Christopher Petzold
(2)
,
Tune Wulff
(1)
,
Sarah O’connor
(4)
,
Vincent Courdavault
(3)
,
Michael Jensen
(1)
,
Jay Keasling
(1, 2, 5)
Lea Hansen
- Fonction : Auteur
- PersonId : 1210293
- ORCID : 0000-0002-8454-312X
Konrad Viehrig
- Fonction : Auteur
- PersonId : 1210294
- ORCID : 0000-0003-0992-3904
Elena Carrasquer-Alvarez
- Fonction : Auteur
- PersonId : 1210295
- ORCID : 0000-0001-9292-9631
Ling Chen
- Fonction : Auteur
- PersonId : 1210296
- ORCID : 0000-0001-6341-6191
Emily Stander
- Fonction : Auteur
- PersonId : 803568
- ORCID : 0000-0002-8382-0631
Christopher Petzold
- Fonction : Auteur
- PersonId : 1210297
- ORCID : 0000-0002-8270-5228
Sarah O’connor
- Fonction : Auteur
- PersonId : 1210298
- ORCID : 0000-0003-0356-6213
Vincent Courdavault
- Fonction : Auteur
- PersonId : 753009
- IdHAL : vincent-courdavault
- ORCID : 0000-0001-8902-4532
- IdRef : 095144439
Michael Jensen
- Fonction : Auteur
- PersonId : 1210299
- ORCID : 0000-0001-7574-4707
Jay Keasling
- Fonction : Auteur
- PersonId : 1210300
- ORCID : 0000-0003-4170-6088
Résumé
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 vincristine 1 . 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 scale 2,3 . 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.